Tuesday, December 29, 2009

Tuj-1-Neuronal Differentiation Marker

Our Tuj-1 antibodies are widely used and frequently referenced in customer publications. They are proven markers for Neural Progenitor and Neuronal Differentiation. Here's the latest reference:

...Tuj 1 (Neuron-specific class III beta-tubulin)-Mouse (MO15013, Neuromics Antibodies, Edina, MN)...

Immunofluorescence Method:

Cells grown on coverslips were fixed for 5 min in 4% paraformaldehydecontaining 4% sucrose in phosphate buffer saline (PBS) at 37ºC. Cells were then permeabilized with 0.2% Triton X-100 in PBS during 5 min at room temperature. After blocking (5% bovine serum albumin in PBS for 1 h), cells were incubated with the corresponding primary antibodies, and immunoreactivity was detected with the suitable fluorophore-conjugated secondary antibody before mounting on slides with Mowiol4-88 (Harland Co., UK). Confocal images were acquired using an inverted Leica TCS SP5 laser confocal microscope with a 63X Plan- Achromatic oil immersion objective and processed with LAS AF Leica Application Suite and Adobe Photoshop CS2 (Adobe Systems
Inc., CA). All images correspond to the projection of sections from a ~50μm z-stack, except for colocalization analysis where they correspond to 0.5-0.7μm single sections.

Image: Rat hippocampal neurons were fixed at 1.5 DIV and immunostained for the neuronal marker βIII-Tubulin/Tuj1 (blue).

Thursday, December 24, 2009

Opioid Receptors and Depression

Tricyclic antidepressants (TCAs) have been reported to interact with the Opioid Receptor system, but their pharmacological activity at opioid receptors has not yet been elucidated.

We would like to share a recent publications that sheds more light on the mystery. Our purified rabbit ployclonal phosphoERK1/2 Antibody is also referenced.

Pierluigi Onali, Simona Dedoni and Maria C. Olianas. Direct Agonist Activity of Tricyclic Antidepressants at Distinct Opioid Receptor Subtypes. JPET January 2010 vol. 332 no. 1 255-265 .

At the cloned μ-opioid receptor, TCAs showed low affinity and no significant agonist activity. These results show that TCAs differentially regulate opioid receptors with a preferential agonist activity on either δ or κ subtypes and suggest that this property may contribute to their therapeutic and/or side effects.

Related Reagents:
phosphoERK1/2 (Rabbit MAb)
Immune Response Research Antibodies
Immune Response Research Proteins
Neurotrophins and Growth Factor Antibdodies
Neurotrophin Proteins
Pain and Inflammation Research Antibodies

Friday, December 11, 2009

Hope for Stroke Victims-Transplanting STEMEZ hNP1 Cells

In a recent publication (Jen et al., 2009) Neuromics'/ArunA’s STEMEZTM human neural progenitor (hNP1) cells when injected (sterotaxic) into a rat stroke model produced significant beneficial results. The hNP1 cells reduced the infarct area by 50% and were positive for neuronal marker proteins, cleaved caspase-3 and 40% of the cells showed spontaneous action potentials and excitatory postsynaptic currents measured by patch clamp recordings at 8 weeks post hNP1 cell injections. The treated rats had improves cognitive and sensorimotor functions between four to nine months post injection. These Sprague–Dawley rats were not immunosuppressed.

Kunlin Jin, XiaoOu Mao, Lin Xie, Veronica Galvan, Bin Lai, Yaoming Wang, Olivia Gorostiza, Xiaomei Wang and David A Greenberg. Transplantation of human neural precursor cells in Matrigel scaffolding improves outcome from focal cerebral ischemia after delayed postischemic treatment in rats. Journal of Cerebral Blood Flow & Metabolism advance online publication 14 October 2009; doi: 10.1038/jcbfm.2009.219.

STEMEZ(TM) hNP1 Human Neural Progenitors Discovery Kit
Other Reagents:
STEMEZ(TM) hN2 Human Neurons Discovery Kit
All Stem Cell Research Reagents
Primary Neurons and Astrocytes

Wednesday, December 09, 2009

New Mouse Monoclonal GFAP Antibody

Check it out!

GFAP Antibody

Excelent marker for human astrocyte intermediate filaments in the central nervous system. It has also been detected in the glial cells of the enteric nervous system and some Schwann cells in the peripheral nervous systems.

Posted using ShareThis

Saturday, December 05, 2009

Blood Pressure, Transient Receptor Potential Vanilloid 1 Receptors and Baroreceptors

Dr. Dr. Hui-Lin Pan (Department of Anesthesiology) and his team at M. D. Anderson Cancer Center has been a loyal user of our TRPV1 (VR1) since the early days of Neuromics' existence. We appreciate their continuing to use our reagent in their research.

Here's yet another publication:

Hao Sun, De-Pei Li, Shao-Rui Chen, Walter N. Hittelman and Hui-Lin Pan. Sensing of Blood Pressure Increase by Transient Receptor Potential Vanilloid 1 Receptors on Baroreceptors. doi:10.1124/jpet.109.160473

...VR1 C-terminus (TRPV1), dilution 1:1000, Neuromics...

Related Reagents:
All TRP Antibodies
Pain and Inflammation Research Antibodies
Neurotransmission -Neurotransmission Research Antibody Categories

Friday, November 13, 2009

NPY Y2R and IHC-Mouse Distal Colon

Lixin Wang, Guillaume Gourcerol, Pu-Qing Yuan, S. Vincent Wu, Mulugeta Million, Muriel Larauche, and Yvette Taché. Peripheral peptide YY inhibits propulsive colonic motor function through Y2 receptor in conscious mice. Am J Physiol Gastrointest Liver Physiol (November 5, 2009). doi:10.1152/ajpgi.00349.2009.
...Note: Excellent IHC staining of myenteric plexus and submucosal (mouse distal colon) tissue-Free floating submucosal and LMMP whole mounts of both proximal and distal colon from 3 naïve mice were treated in 10% normal goat serum each for 30 min, and followed by incubation with polyclonal rabbit anti- NPY Y2 Receptor diluted at 1:1,000 (Neuromics, Inc., Edina, MN)...

Related Antibodies to Consider:
NPY Y2 Receptor-C/N Terminus
NPY Y1 Receptor
All Neuropeptide and Neuropeptide Receptor Antibodies
Pain and Inflammation Research Antibodies
Diabetes and Obesity Research Antibodies

More Pubs Referencing Neuromics' mGluRs

Jakob S. Satz, Alisdair R. Philp, Huy Nguyen, Hajime Kusano, Jane Lee, Rolf Turk, Megan J. Riker, Jasmine Hernández, Robert M. Weiss, Michael G. Anderson, Robert F. Mullins, Steven A. Moore, Edwin M. Stone, and Kevin P. Campbell. Visual Impairment in the Absence of Dystroglycan. J. Neurosci., Oct 2009; 29: 13136 - 13146 ; doi:10.1523/JNEUROSCI.0474-09.2009.
....anti-mGluR6 (Neuromics)...

Lasani S. Wijetunge, Sally M. Till, Thomas H. Gillingwater, Cali A. Ingham, and Peter C. Kind. mGluR5 Regulates Glutamate-Dependent Development of the Mouse Somatosensory Cortex. The Journal of Neuroscience, December 3, 2008, 28(49):13028-13037; doi:10.1523/JNEUROSCI.2600-08.2008.
...Western blotting was performed as mentioned above and membranes were probed with antibodies against mGluR5 (1:4000, Neuromics)...
Posted using ShareThis

Friday, November 06, 2009

gp130, IL-6 and Expression and Neuropathic Pain

We wanted to post yet another publication referencing use of one of our Pain and Inflammation Research Antibodies. Here the researchers used our Mouse Monoclonal gp130/CD130 for Immunohistochemistry.

Manfred Andratsch, Norbert Mair, Cristina E. Constantin, Nadja Scherbakov, Camilla Benetti, Serena Quarta, Christian Vogl, Claudia A. Sailer, Nurcan Üceyler, Johannes Brockhaus, Rudolf Martini, Claudia Sommer, Hanns Ulrich Zeilhofer, Werner Müller, Rohini Kuner, John B. Davis, Stefan Rose-John, and Michaela Kress. A Key Role for gp130 Expressed on Peripheral Sensory Nerves in Pathological Pain. J. Neurosci., Oct 2009; 29: 13473 - 13483 ; doi:10.1523/JNEUROSCI.1822-09.2009

The results suggest that gp130 expressed in sensory nerves not only mediates chronic inflammatory pain, but also contributes significantly to complex interactions between immune cells, tumor cells, and nerves in the context of cancer-evoked pain. Moreover, we identify IL-6 activating gp130, Gab1/Gab2, PI3K, and PKC- and regulating TRPV1 as a key mechanism linking cytokine release to sensitization of pain-sensing nerves.

Related Links
Immune Response
Opioid Neuropeptides
Opioid Receptors
Neurotransmissiom Research Antibodies
GPCRs, Ligand Gated Ion Channels, Biogenic Amines and more
Purinergic Receptors
TRPV (Vanilloid); TRPM; TRPA and TRPCs
i-Fect Transfection Kit
gene silencing of DOR, NaV1.8 tetrodotoxin-resistant sodium channel, NTS2 and more in-vitro and in vivo
Primary Neurons and Astrocytes
Primary human, rat and mouse neurons and astrocytes by Category

Monday, October 19, 2009

NSE and TUJ-1 and Parkinson's Disease Research

I would like to thank Meghan Coakley from University College Cork for alerting me to a new publication referencing our Chicken NSE (Neuron-Specific Enolase) and Tuj 1 (Neuron-specific class III beta-tubulin) antibodies.

Here's her feedback: "Just letting you know we published our paper using the beta-III-Tubulin and NSE antibodies you supplied to us – Timmons et al., Neuroscience Letters, Oct 1, 2009 [Epub ahead of print]. The antibodies were excellent and I’m sure we’ll be using Neuromics again in the future."

Timmons S, Coakley MF, Moloney AM, O' Neill C. Akt signal transduction dysfunction in Parkinson's disease. Neurosci Lett. 2009 Oct 1. [Epub ahead of print].

Abstract: Significant attention has been drawn to the potential role of defective PI3-kinase-Akt (PKB) signalling in Parkinson's disease (PD) neurodegeneration and to the possibility that activation of Akt may provide neuroprotection in PD. However, little knowledge exists on the integrity of the Akt system in PD. Results of the present study show diminished levels of both total and active phospho(Ser473)-Akt in the brain in PD. This was evident by western blot analysis of midbrain fractions from PD compared to non-PD control brain, but more specifically by immunofluorescence microscopy of the substantia nigra pars compacta (SNpc). Here, double immunofluorescence microscopy found Akt and phospho(Ser473)-Akt to be expressed at high levels in tyrosine hydroxylase (TH) immunopositive dopaminergic neurons in control human brain. Selective loss of these neurons was accompanied by a marked decrease of Akt and phospho(Ser473)-Akt expression in the PD brain, however Akt and active phospho(Ser473)-Akt are still evident in degenerating dopaminergic neurons in the disease. This suggests that it may be possible to target neuronal Akt in advanced PD. Converse to the marked loss of neuronal Akt in PD, increased Akt and phospho(Ser473)-Akt levels were observed in small non-TH positive cells in PD SNpc, whose increased number and small nuclear size indicate they are glia. These findings implicate defective Akt as a putative signalling pathway linked to loss of dopaminergic neurons in PD.

Other Reagents to Consider:
Tuj 1 (Neuron-specific class III beta-tubulin)-Mouse Monoclonal
Neuron/Glial Marker Antibodies
Neurotrophins-Neuron/Glial Marker Proteins
Neurodegenerative Disease Research Antibodies
Neurodegenerative Disease Research Proteins
Stem Cell Research Reagents

Friday, October 09, 2009

MMP-9 Squared

We are feverishly working on adding reagents for studying Autoimmunity and Immune Response. In this context, we wanted to alert you to 2 new publications referencing our Matrix Metalloproteinase 9 (MMP-9) antibody.

Zhi-Yuan Zhang, Zhiren Zhang, Caroline Zug, Barbara Nuesslein-Hildesheim, David Leppert and Hermann J. Schluesener. AUY954, a selective S1P1 modulator, prevents experimental autoimmune neuritis. doi:10.1016/j.jneuroim.2009.09.010.

Experimental autoimmune neuritis (EAN) is a T cell-mediated autoimmune inflammatory demyelinating disease of the peripheral nervous system and an animal model of human
inflammatory demyelinating polyradiculoneuropathy. AUY954, which targets selectively the sphingosine-1-phosphate receptor 1 (S1P1), is known to sequester lymphocytes into secondary lymphoid tissues. In EAN rats, AUY954 greatly prevented paraparesis if administrated from the day of immunization. T cell, B cell, and macrophage infiltration, inflammatory demyelination, and local expression of interleukine-17 and matrix metalloproteinase-9 in sciatic nerves of EAN rats were significantly decreased by AUY954 treatment. Therefore, S1P1 modulation might be a potential treatment option for inflammatory neuropathies.

Image: MMP-9 expression in the sciatic nerves of control (A) vs AUY954 (B) treated rats.

Yasuki Yasuda, Yoko Matsumura, Kazuki Kasahara, Noriko Ouji, Shigeki Sugiura, Keiichi Mikasa, and Eiji Kita. Microbial exposure early in life regulates airway inflammation in mice after infection with Streptococcus pneumoniae with enhancement of local resistance. Am J Physiol Lung Cell Mol Physiol, Sep 2009; doi:10.1152/ajplung.00193.2009

...with 10% milk in TBST (10mM Tris, 0.15M NaCl, 0.1% Tween-20), followed by probing with goat anti-mouse MMP-9 antibodies (Ab) (Neuromics Antibodies, Edina, MN, GT15020; 1:5000)...

Related Reagents:

CD and Cell Surface Markers
Heat Shock Proteins-New
Matrix Metalloproteinases (MMPs)
ILs, CCRs, CXCs and STATs
Other Immune Response Antibodies

Immune Response Research Proteins

Apoptosis Research Reagents

Wednesday, September 30, 2009

Rock Solid Tuj-1

The feedback from customers on our Tuj 1 (Neuron-specific class III beta-tubulin) is that it works!

This is confirmed by the growing list of references in key publications. Here's the latest:

S A Sakowski, S B Heavener, J S Lunn, K Fung, S S Oh, S K Spratt, N D Hogikyan and E L Feldman. Neuroprotection using gene therapy to induce vascular endothelial growth factor-A expression. Gene Therapy advance online publication 3 September 2009; doi: 10.1038/gt.2009.111.

...TUJ1 (Neuromics, Edina, MN, USA). ...

Tuj 1 (Neuron-specific class III beta-tubulin)
Related Reagents:
Other Reagents to Consider:
Stem Cell Reagents
Neuron/Glial Markers

Monday, September 28, 2009

Prodynorphin at Work

We would like to Dr. Andrew Todd for sharing this excellent IHC image using our Guinea Pig ProDynorphin (rat) antibody.

Image: Staining of adult rat spinal cord.

The tissue is perfusion-fixed (4% freshly prepared formaldehyde) adult rat spinal cord, reacted overnight with the PPD at 1:1000 and then o/n in Alexa488 secondary (raised in donkey, Invitrogen, 1:500).

The confocal image stack was taken through a 60x oil lens (Bio-Rad Radiance confocal) - pixel size is 0.196 micrometre and this is a projection of 10 confocal optical sections at 0.5 micrometre z-spacing.
Customer Publications
Related Reagents:
proDynorphin (guinea pig)
Opioid Receptors
Pain and Inflammation Antibodies

Thursday, September 10, 2009

TRPV1 Antibodies in Action

We would like to thank Dr. Fletcher White, LUMC, for Sharing this data with us:

Images: The CCR2 chemokine receptor colocalized with IB4 and TRPV1, markers of nociceptive neurons, after injury. A) Many lumbar DRG neurons in vehicle-treated rat sensory neurons were positive for IB4, a neuronal phenotype that distinguishes some C-fiber nociceptors (red cells), however there was no expression of the CCR2 protein. B) After perineural gp120/hCD4 treatment, CCR2 protein expression (green cells) was upregulated, and co-localized with IB4. C) Both gp120/hCD4 and ddC treatment resulted in an upregulation of CCR2 expression (green cells) in many small and medium diameter neurons. Again, CCR2 co-localized in a number of IB4 positive cells. D) The TRPV1 channel is present on many nociceptive neurons and is involved in the neuropathic pain mechanism. Under normal conditions, TRPV1 was expressed in neurons (red cells). E) After gp120/hCD4 treatment, CCR2 expression is upregulated (green cells) and colocalized with TRPV1. F) After the combination of gp120/hCD4 and ddC treatments, again CCR2 was upregulated to a similar degree as gp120/hCD4 treatment alone and exhibited some colocalizations with TRPV1.
Scale Bar: 100um.

We would also like to share recent publications referencing our TRPVs:

Hao Sun, De-Pei Li, Shao-Rui Chen, Walter Hittelman, and Hui-Lin PanSensing of Blood Pressure Increase by Transient Receptor Potential Vanilloid 1 Receptors on BaroreceptorsJ. Pharmacol. Exp. Ther., Sep 2009; doi:10.1124/jpet.109.160473 ......guinea pig anti-TRPV1, dilution 1:1000, Neuromics, Minneapolis, MN) and secondary antibody...guinea pig anti-VR1 C-terminus (TRPV1), dilution 1:1000,Neuromics; and rabbit anti-NF200, dilution 1:100...guinea pig anti-TRPV1, dilution 1:1000, Neuromics) for 2 hr at room temperature and

overnight......Kenjiro Matsumoto, Emi Kurosawa, Hiroyuki Terui, Takuji Hosoya, Kimihito Tashima, Toshihiko Murayama, John V. Priestley, and Syunji HorieLocalization of TRPV1 and contractile effect of capsaicin in mouse large intestine: high abundance and sensitivity in rectum and distal colonAm J Physiol Gastrointest Liver Physiol, Aug 2009; 297: G348 - G360. ......TRPV1 antibody (mouse TRPV1 C-terminus; Neuromics, Minneapolis, MN) were 1:60,000 for rectum...different anti-TRPV1 antibodies (1:60,000, Neuromics, and rat TRPV1 COOH-terminus, 1:1,000...experiments, the antibody (1:60,000; Neuromics) was preincubated with 10 uM of the corresponding......

Featured Reagents:
VR1 N-Terminus (TRPV1)
VR1 C-terminus (TRPV1)
VR1 C-Terminus (TRPV1) - mouse specific
Related Reagents
VR1 (TRPV1)-Goat
VR like-3 (TRPV3)
All TRPV (Vanilloid); TRPM; TRPA and TRPCs
Pain and Inflammation Antibodies

Friday, August 28, 2009

Tracking STEMEZ hNP1 Progenitor Cell Fate

We would like to promote an important new discovery using our STEMEZ(TM) hNP1 Human Neural Progenitors Discovery Kit.

Human embryonic stem cell–derived neural progenitors (NP) present an important tool for understanding human development and disease. Optimal utilization of NP cells, however, requires an enhanced ability to monitor these cells in vitro and in vivo. Here we report production of the first genetically modified self-renewing human embryonic stem cell–derived NP cells that express fluorescent proteins under constitutive as well as lineage-specific promoters, enabling tracking and monitoring of cell fate. Nucleofection, transfection, and lentiviral transduction were compared for optimal gene delivery to NP cells. Transduction was most efficient in terms of transgene expression (37%), cell viability (39%), and long-term reporter expression (>3 months). Further, the constitutive gene promoters, cytomegalovirus, elongation factor 1α, and ubiquitin-C, exhibited comparable silencing (20–30%) in NP cells over a 2-month period, suggesting their suitability for long-term reporter expression studies. Transduced NP cells maintained their progenitor state and differentiation potential, as demonstrated by expression of endogenous NP markers and neuronal markers after differentiation. We also detected reporter expression in astrocytes generated from NP cells transduced with an astrocyte-specific gene promoter, glial fibrillary acidic protein, demonstrating the usefulness of this approach. The genetically manipulated NP cells described here offer great potential for live cell–tracking experiments, and a similar approach can as well be used for expression of proteins other than reporters.

Related Publication:
Sujoy K. Dhara, Brian A. Gerwe, Anirban Majumder, Mahesh C. Dodla, Nolan L. Boyd, David W. Machacek, Kowser Hasneen, Steven L. Stice. Genetic Manipulation of Neural Progenitors Derived from Human Embryonic Stem Cells. Tissue Engineering Part A. -Not available-, ahead of print. doi:10.1089/ten.tea.2009.0155

Neuromics' Stem Cell Markers Referenced:
Musashi-1, Rabbit, 1:100 , Catalog#:RA14128
Nestin, Mouse 1:500, Catalog#: MO15012
Tuj 1 (Neuron-specific class III beta-tubulin), Mouse, 1:500, Catalog#: MO15013
TH, Chicken, 1:80, Catalog#: CH23006

Wednesday, August 19, 2009

STEMEZ hNP1 Neural Progenitors Now Available

We are pleased to announce the introduction of our STEMEZ TMhNP1 Human Neural Progenitors Discovery Kit. The kit was developed by our partner, ArunA Biomedical.

STEMEZTM hNP1 Human Progenitors are fully differentiated are derived as adherent cells from hESC WA09 line. The cells are shipped frozen in a vial with 1 x 106 cells; once thawed they should be immediately plated on a Matrigel (or other suitable extracellular matrix protein)-coated dish and maintained in the accompanying serum-free medium. The neurons should be used within 14 days of thawing. These cells have the capabilities to:
  • Display immunoreactive properties consistent with neural precursors and can be maintained in a proliferative state in monolayer cultures.
  • Differentiate under serum-free conditions into neuronal phenotypes with functionally responsive transmitter receptors in vitro .
  • Maintain multiple neural phenotypes in long term serum-free culture.
  • Can be directed to alter phenotypic characteristics under different media conditions.

Applications: Gene Expression Analysis Western blotting, Flow Cytometry, Immunocytochemistry, FACS sorting, DNA Microarray, RT-PCR, Neurite Outgrowth assays, FLIPR calcium assays, cytotoxicity, and second messenger signaling.

Images: STEMEZTM hNP1 Human Neural Progenitor Cells are grown as monolayers (A), are karyotypically normal (B) and express NSC markers, Nestin Mouse and Sox-2 (C, D, E). Nuclei of the cells were visualized with DAPI (blue). The Sox-2 transcription factor is co-localized with the DAPI (blue) staining in the nucleus (F).


Note: STEMEZTM NP1 Progenitors and hN2 Human Neurons can be genetically manipulated with GFP reporters without altering the function and differentiation potential of these cells. In contrast to many stem cell sources, the reporter genes are not readily silence in the neural cultures. See: doi:10.1089/ten.tea.2009.0155.

DNA fingerprint cells: The loci match the DNA fingerprint pattern for the H9 (NIH designation, WA09) HESC line as published in http://stemcells.nih.gov/research/nihresearch/scunit/.

Viral tests cells: This lot was derived from the H9 hESC line that has been tested for Hepatitis B, Hepatitis C, HIV-1, HIV-2, HTLV-I/II, HSV1, HSV2, EBV, and CMV. The H9 cell line has been tested and shown to be negative. (Tests performed by GIVF Laboratories).

Related Reagents:

Sunday, August 16, 2009

Link Between Inflammation and Tau Pathology in Alzheimer's Disease

Providing reagents for Alzheimer's Disease Researchers is a strategic focus area for us. This includes making a variety of Primary Neuronal Cultures available to these researchers.

We evolve the strategy based on Researcher input and how the cultures are being referenced in publications. This reference is especially satisfying because of the vangaurd findings:

Lisette T. Arnaud, Natura Myeku and Maria E. Figueiredo-Pereira. Proteasome–caspase–cathepsin sequence leading to tau pathology induced by prostaglandin J2 in neuronal cells. Journal of Neurochemistry. Volume 110 Issue 1, Pages 328 - 342.

"Our data suggest a potential sequence of events triggered by the neurotoxic product of inflammation PGJ2 leading to tau pathology. The accumulation of Ub proteins is an early response. If cells fail to overcome the toxic effects induced by PGJ2, including accumulation of Ub proteins, apoptosis kicks in triggering caspase activation and tau cleavage, the clearance of which by cathepsins could be compromised culminating in tau pathology. Our studies are the first to provide a mechanistic link between inflammation and tau pathology."

Featured Reagent:
E18 Primary Rat Cortical Neurons

Related Reagents:

Images: Neural phenotypes derived from hN2 cell lines. (A) Phase contrast image of differentiated culture. (B) Network including post-mitotic motoneurons (HB9). (C) Cholinergic neuron. (D) Tuj-1 positive cells that are DAT-positive (dopamine transporter; closed arrow) and DAT-negative (open arrow). (E) Gabaergic neurons, inset illustrates GABA in axon, but not the dendrites (arrow).
We will continue to post "whats new" in AD research and potential uses of our reagents.

Saturday, August 15, 2009

survival of efferent synapses on mammalian outer hair cells.

Dr Douglas Vetter (Tufts University School of Medicine) and team recently published a study that amplifies the understanding of the development, function and maintenance of auditory system function.

Their data strongly suggest that hair cell responses induced and/or modulated by Olivocochlear (OC) activation are necessary for the survival of OC innervation and that these responses must involve SK2-mediated hyperpolarization.

It also includes an excellent image of Olivocochlear fibers degeneration in SK2−/− mice. One of the makers used was our Tuj 1 (Neuron-specific class III beta-tubulin).

Vidya Murthy, Stéphane F. Maison, Julián Taranda, Nadeem Haque, Chris T. Bond , A. Belén Elgoyhen, John P. Adelman, M. Charles Liberman, Douglas E. Vetter. SK2 channels are required for function and long-term survival of efferent synapses on mammalian outer hair cells. Molecular and Cellular Neuroscience 40 (2009) 39–49
...TuJ-1 (class III β-tubulin, Neuromics, Northfield, MN; cat. # MO15013) and processedwith Oregon Green labeled secondary antibodies (Molecular Probes/InVitrogen) for confocal microscopy...

Other Pubs Referencing Neuromics' Tuj 1 (Neuron-specific class III beta-tubulin):
Yoshifumi Saisho, Paul E. Harris, Alexandra E. Butler, Ryan Galasso, Tatyana Gurlo,1 Robert A. Rizza, and Peter C. Butler. Relationship between pancreatic vesicular monoamine transporter 2 (VMAT2) and insulin expression in human pancreas. J Mol Histol. 2008 October; 39(5): 543–551. Published online 2008 September 13. doi: 10.1007/s10735-008-9195-9.
Sujoy K. Dhara, Kowser Hasneen, David W. Machacek, Nolan L. Boyd, Raj R. Rao, Steven L. Stice (2008). Human neural progenitor cells derived from embryonic stem cells in feeder-free cultures. Differentiation 76 (5) , 454–464 doi:10.1111/j.1432-0436.2007.00256.x
...NES (1:100, Neuromics, Edina, MN), MSI1 (1:100, Neuromics), Tuj1 (1:500, Neuromics)...
D. Conte, V. Lall, G. Dobson, S. Deuchars, J. Deuchars. Cerebrospinal fluid contacting neurones in the spinal cord of the mouse and rat: small cells with a big purpose? University of Leeds (2008) Proc Physiol Soc 10 PC45
...Preliminary immunohistochemical data supports the concept that the CSFcNs are immature neurones, since they express neuronal markers Tuj1 (Neuromics) ...

Saturday, August 08, 2009

ENS Development Markers

This is an excellent study on the developing Enteric Nervous Stem (ENS). This system is often referred to as the "second brain".

Despite this, neurogenesis has been much less studied in the ENS than in the brain. Understanding how neurons are formed in the gut is the foundation for finding cures for ENS disorders. The key finding here is that the ability of 5-HT4 receptors to unmask a regulation of enteric neurogenesis in adult animals suggests that the mature ENS is capable of an unexpected degree of plasticity (potentially good new for discovering therapies for ENS related disorders).

Min-Tsai Liu, Yung-Hui Kuan, Jingwen Wang, René Hen, and Michael D. Gershon. 5-HT4 Receptor-Mediated Neuroprotection and Neurogenesis in the Enteric Nervous System of Adult Mice. The Journal of Neuroscience, August 5, 2009, 29(31):9683-9699; doi:10.1523/JNEUROSCI.1145-09.2009.

More details @ http://www.anxietyinsights.info/serotonin_the_gut_and_neurogenesis.htm

On a side note, this publication is rich with excellent ICC and Western Blot images of a variety of Neurogenesis Markers. This include referencing of 4 of our markers:

GFAP-CH22102-ICC Dilution 1:2000
Musashi-1-RA14128-ICC Dilution 1:100 WB Dilution 1:1000
Neurofilament NF-H-CH22104-ICC Dilution 1:2000
S100B-RA25022-ICC Dilution 1;1000

Tuesday, August 04, 2009

Spinal Cord Injury Repair

Neuromics has featured an overview of Dr. Matt Ramer's Research on Spinal Cord Injury and "Finding Fixes for Injured Nerves" on our Neuroscience News Blog.

We wanted to share a recent publication authored by Dr. Mark Tuszynski and his team of researchers at USCD. Here's the good news:

"NT-3 expression in the correct target led to reinnervation of the nucleus gracilis in a dose-related fashion, whereas NT-3 expression in the reticular formation led to mistargeting of regenerating axons. Axons regenerating into the nucleus gracilis formed axodendritic synapses containing rounded vesicles, reflective of pre-injury synaptic architecture. Thus, we report for the first time, to the best of our knowledge, the reinnervation of brainstem targets after SCI and an essential role for chemotropic axon guidance in target selection"

Laura Taylor Alto, Leif A Havton, James M Conner, Edmund R Hollis II, Armin Blesch & Mark H Tuszynski. Chemotropic guidance facilitates axonal regeneration and synapse formation after spinal cord injury. Nature Neuroscience. Published online: 2 August 2009 doi:10.1038/nn.2365.

Featured Neuromics Reagent
Related Reagents:
Neurotrophins and Growth Factor Antibodies
Neuron/Glial Marker Antibodies
Neurotrophins-Neuron/Glial Marker Proteins
Stem Cell Reagents

Here's to finding the cure!

Sunday, July 26, 2009

STEMEZ hN2 Human Neurons-Data

Neuromics rolled out STEMEZTM hN2 Human Neurons Discovery Kits several months ago.

Applications for these include: cellular model studies, high content screening, developmental studies, RNAi studies and genetic manipulation.

Drilling down further, I am pleased to present Electro-physiology and related data generated by Aruna and collaborators: hN2 Cells-Electro Phys Data Supplement

hN2-Whole Cell Voltage Clamp
Figure. hN2 cells can produce inward currents that generate action potentials. (A) Isolated hN2 with significant neurite growth 1 week after plating . This cell was subjected to whole cell voltage clamp utilizing a potassium gluconate based intracellular solution. (B) Voltage gated inward and outward currents were elicited from this cell with depolarizing voltage steps. (C) Inward currents from another cell (potassium gluconate intracellular) were abolished by local application of 1 µM tetrodotoxin (red trace) while outward currents remained. Inward current recovered as TTX washed out of the region (green trace). (D) A different cell which exhibited voltage activated inward currents that inactivated in response to a 50 ms prepulse at different membrane potentials. The experiment was done 27 days after the removal of bFGF. A cesium gluconate based intracellular solution was used for this experiment to block outward potassium currents. The membrane potential for half maximal inactivation by standard Boltzman fitting (red line) was -40.1 mV with a slope of 4.7. (E) Recovery from fast inactivation utilizing a paired pulse protocol in the same cell as C. The single exponential time constant for recovery of inactivation was 1.7 ms (red line). (F) A different cell which elicited an overshooting action potential upon current injection under whole cell current clamp utilizing a potassium gluconate based intracellular solution. Inset: Response of the same cell under voltage clamp to a change in membrane potential from -80 mV to -10 mV elicited a peak current of 457 pA. Scale bars for inset: 5 ms, 0.2 nA.

Thursday, July 23, 2009

Stem Cell Markers

Yet more confirmations of the potency of our Stem Cell Markers.

In this study the authors demonstrate a between SSCs and RET.

ETV5 is expressed beginning in fetal Sertoli cells and can potentially have effects on both neonatal Sertoli and germ cells. In addition, ETV5 has critical effects on neonatal spermatogonial proliferation, which may involve impaired signaling through the RET receptor.

Gaurav Tyagi, Kay Carnes, Carla Morrow, Natalia V. Kostereva, Gail C. Ekman, Daryl D. Meling,
Chris Hostetler, Michael Griswold, Kenneth M. Murphy, Rex A. Hess, Marie-Claude Hofmann and Paul S. Cooke. Loss of Etv5 Decreases Proliferation and RET Levels in neonatal Mouse Testicular Germ Cells and Causes an Abnormal First Wave of Spermatogenesis.

Images: Ret IHC in WT and Etv5-/- testes (n=4). The intensity of RET staining per spermatogonia is markedly reduced in Etv5-/- testis (B) even though the number of cells staining for RET are comparable to the WT (A) testis. Insets are higher magnification of spermatogonia showing differences in staining intensity. Bars = 50 μm in A and B, insets = 10 μm.

Patent:Neuronal progenitors from feeder-free human embryonic stem cell culture.
Inventor: Stice, et al.
Date Issued: May 12, 2009
Application: 11/243,819
Stice; Steve (Athens, GA)Shin; Soojung (Baltimore, MD)Dhara; Sujoy (Athens, GA)
University of Georgia Research Foundation, Inc. (Athens, GA)Include use of Nestin antibody

Stem Cell Antibodies
Stem Cell Research Proteins
Neural Stem Cells and Media
Neuroprogenitor Neurosphere Tissue-NEW!
Neuroprogenitor Neurosphere tissue is provided live, unseparated, fresh from E18 rat cortex/hippocampus including subventricular zone.
Expansion/Differentiation Kits
Related Reagents:
Neurotrophin and Growth Factor Antibodies
Neurotrophin Proteins

Friday, June 26, 2009

Hes-1 and Develpmental Neurobiology

Developmental Neurobiology is a core area of focus for Neuromics. Our goal is to provide a growing catalog of research proven reagents to Researchers. Their work is key to unlocking root causes of developmental related diseases and defects.

We continue to see use of our reagents in publications by leading labs. Here's a new publication by Dr. Paul Trainor and his team at Stower's Institute referencing use of our HES-1 antibody:

Sunday, June 14, 2009

Potent Stem Cell Markers

The potency of our Stem Cell Research Reagents is confirmed by the growng list of publication referencing them.

Here're the latest:

Saishu Yoshida, Takako Kato, Takao Susa, Li-yi Cai, Michie Nakayama, Yukio Kato. PROP1 coexists with SOX2 and induces PIT1-commitment cells. Biochemical and Biophysical Research Communications, Volume 385, Issue 1, 17 July 2009, Pages 11-15
... SOX2 (1:500 dilution, Neuromics, Edina, MN, USA)...

Saravanan Karumbayaram, Bennett G. Novitchb, Michaela Patterson, Joy A. Umbach, Laura Richter, Anne Lindgren, Anne E. Conway, Amander T. Clark, Steve A. Goldman, Kathrin Plath, Martina Wiedau-pazos, Harley I. Kornblum, William E. Lowry. Directed Differentiation of Human-Induced Pluripotent Stem Cells Generates Active Motor Neurons. Stem Cells Vol. 27 No. 4 April 2009, pp. 806 -811. doi:10.1002.
... mouse anti-Nestin (Neuromics)...

Patrizia Rubini, Javorina Milosevic, Johannes Engelhardt, Mahmoud Al-Khrasani, Heike Franke, Attilla Heinrich, Beata Sperlagh, Sigrid C. Schwarz, Johannes Schwarz, Wolfgang Nörenberg and Peter Illes. Increase of intracellular Ca2+ by adenine and uracil nucleotides in human midbrain-derived neuronal progenitor cells. Cell Calcium. Volume 45, Issue 5, May 2009, Pages 485-498.
...rabbit anti-P2Y2 (1:1000; Neuromics, Edina, MN, USA...

Thursday, May 07, 2009

Detecting Apoptosis in Real Time

Kudos to Drs. Aaron Thomas and Carol Erickson for their novel use of our Magic Red™ Real Time! Apptosis Kits. Here's the related publication:

Aaron J. Thomas and Carol A. Erickson. FOXD3 regulates the lineage switch between neural crest-derived glial cells and pigment cells by repressing MITF through a non-canonical mechanism. Development, Apr 2009; doi:10.1242/dev.031989.
...Apoptosis was assayed using Magic Red Caspases 3&7 reagent (Neuromics)...

Related Reagents:
FLIVO™ Polycaspase Live!, in vivo Apoptosis Kits-New
FLICA™ in vitro Caspase Kits
Fast!-Use Caspase kits to quantitate apoptosis via active caspases in whole, living cells. These kits do not use ELISA or any antibodies for detection
FLISP™ Serine Protease Detection Kits
Measure chymotrypsin-like proteaseactivation in whole living cells.
MitoPT™ Kits
Quantitate mitochondrial functionality and apoptosis
Apoptosis Research Antibodies
Apoptosis Research Proteins

Thursday, April 02, 2009

RAGE and Pneumonia

We are working with customers and collaborators to strengthen our product offerings for Immune Response Researchers.

An interesting finding on Receptor for Advanced Glycation End Products (RAGE) and response to S. pneumoniae pneumonia infection just crossed our radar. Dr. Marieke A. D. van Zoelen and team published evidence that RAGE plays a detrimental role in the host response to S. pneumoniae pneumonia by facilitating the bacterial growth and dissemination and concurrently enhancing the pulmonary inflammatory and procoagulant response. Data include use of our RAGE-Cat#: GT15030.

Here's the publication and related data:

Marieke A. D. van Zoelen, Marcel Schouten, Alex F. de Vos, Sandrine Florquin, Joost C. M. Meijers, Peter P. Nawroth, Angelika Bierhaus, and Tom van der Poll. The Receptor for Advanced Glycation End Products Impairs Host Defense in Pneumococcal Pneumonia. J. Immunol., Apr 2009; 182: 4349 - 4356.

...Endogenous peroxidase activity was quenched using 1.5% H2O2 in PBS. Primary Abs used were goat anti-mouse RAGE polyclonal Abs (Neuromics), and secondary Abs were biotinylated rabbit anti-goat Abs (DakoCytomation). ABC solution (DakoCytomation) was used as the...

Images: Expression of RAGE in lungs during S. pneumoniae pneumonia. Representative view of a lung from a normal, uninfected Wt mouse (A) displaying ubiquitous expression of RAGE on the surface of endothelium. B, Absence of RAGE positivity in the lung of a RAGE–/– mouse. C and D, Lungs from a Wt mouse 48 h after the inoculation of S. pneumoniae. Arrow indicates bronchial epithelium in healthy lungs (A); asterisk indicates neutrophils in an area with confluent pneumonia (D), both being negative for RAGE staining. RAGE staining: original magnification x10.

Realted Reagents:
RAGE Mouse Mononclonal
Immune Response Antibodies

Immune Response Proteins

Monday, March 30, 2009

Introducing Human Neuron Kits

hN2 Human Neurons Discovery Kit-New

Energize you Research!
Neuromics has formed an alliance with Aruna Biomedical. This Alliance gives us the capabilities to bring you the reliable, robust and highly scalable hN2TMHuman Neurons Discovery Kits.
These kits are designed to reduce basic Neuroscience Research and Drug Discovery timelines. Potential applications include: cellular model studies, high content screening, developmental studies, RNAi studies and genetic manipulation.
Approximate Yield=1,000,000 healthy Neurons.

hN2 Human Neurons Discovery Kit Details

Friday, March 13, 2009

Amyloid Beta and E18 Primary Hippocampal Neurons

We would like to feature an interesting application of our E18 Rat Primary Hippocampal Neurons.

Related Publication:

Karunya K. Kandimalla1, Olenych G. Scott, Smita Fulzele1, Michael W. Davidson, Joseph F. Poduslo. Mechanism of Neuronal versus Endothelial Cell Uptake of Alzheimer's Disease Amyloid β Protein. PLoS ONE 4(2): e4627. doi:10.1371/journal.pone.0004627.

... Rat primary hippocampal (RPH) neurons were isolated from the hippocampii of 18-day-old embryonic Sprague Dawley rat brains (Neuromics, Edina, MN). The hippocampii were dispersed using a fire polished Pasteur pipette and plated on poly-D-lysine (Sigma-Aldrich, St. Louis, MO) coated glass cover slips in B-27 neurobasal medium containing 0.5 mM glutamine and 25 µM glutamate (Invitrogen, Carlsbad, CA). The neuronal cells were grown under 5% CO2 in an incubator maintained at 37°C until differentiation...

Images: A–D: Uptake of fluorescein labeled Aβ40 (F-Aβ40) and Alexa Fluor® 633 labeled transferrin (AF633-Trf), clathrin-mediated endocytosis marker, in rat primary hippocampal (RPH) neurons following 30 min incubation at 37°C. (A) F-Aβ40 uptake; (B) Uptake of AF633-Trf; (C) Superimposition of images A and B; (D) Overlay of fluorescence images on the DIC image of RPH neurons. E–G: Uptake of F-Aβ40 and AF633-Trf in RPH neurons at 4°C. (E) Uptake of F-Aβ40; (F) No significant neuronal uptake of AF633-Trf at 4°C; (G) Superimposition of images D and E on the DIC image of RPH neurons; H–J: Uptake of F-Aβ40 and AF633-Trf in RPH neurons treated with 10 mM Sodium Azide and 50 mM 2-deoxy glucose, agents that are known to deplete cellular ATP. (H) Uptake of F-Aβ40; (I) No significant cellular uptake of AF633-Trf was observed; (J) Superimposition of images H and I on the DIC image of neurons.doi:10.1371/journal.pone.0004627.g010

Saturday, February 28, 2009

TRPV1 with a Twist

Kudos to Dr. Marna Erickson and her team at the University of Minnesota. They recently published excellent work demonstrating a relationship between TRPV1 -EGFR signaling and skin cancer.

Ann M. Bode, Yong-Yeon Cho, Duo Zheng, Feng Zhu, Marna E. Ericson, Wei-Ya Ma, Ke Yao, and Zigang Dong. Transient Receptor Potential Type Vanilloid 1 Suppresses Skin Carcinogenesis.Cancer Res., Feb 2009; 69: 905 - 913.

"TRPV1 interacts with EGFR, leading to EGFR degradation. Notably, the absence of TRPV1 in mice results in a striking increase in skin carcinogenesis. The TRPV1 is the first membrane receptor shown to have a tumor-suppressing effect associated with the down-regulation of another membrane receptor."

...dorsal skin samples (100 mum) were processed and immunostained. For the human skin cancer tissue array, we used anti- TRPV1 -(Neuromics), anti-EGFR (Cell Signaling), and Alexa Fluor 488 and 647-conjugated secondary antibodies. Mouse skin samples were immunostained...

Sunday, February 22, 2009

TRPV1 and Retinal Ganglion Cell Apoptosis

The publications referencing our TRPV (Vanilloid); TRPM; TRPA and TRPC Antibodies keep on coming.

In this February 2009 publication, Dr. David Calkins and his team at Vanderbilt demontrate that Retinal Ganglian Cells express the TRPV1 channel and that TRPV1 activation contributes to their death with exposure to hydrostatic pressure . We also demonstrated that activation of TRPV1 alone was sufficient to induce apoptosis of RGCs.
Rebecca M. Sappington, Tatiana Sidorova, Daniel J. Long, and David J. Calkins. TRPV1: Contribution to Retinal Ganglion Cell Apoptosis and Increased Intracellular Ca2+ with Exposure to Hydrostatic Pressure. Invest. Ophthalmol. Vis. Sci., Feb 2009; 50: 717 - 728.
...we used rabbit anti-mouse TRPV1 IgG (1:500; catalog number RA14113; Neuromics, Edina, MN) against the absolute C terminus of mouse TRPV1 (EDAEVFKDSMAPGEK)...

TRPV1 localization in RGCs of rat retina. (A) Immunocytochemical labeling for TRPV1 shows strong localization in the outer retina and in RGCs (large cell bodies); there is little or no label in smaller displaced amacrine cells of the GCL. Clear examples of amoeboid-shaped cell bodies of microglia cells are indicated (ovals). Right: Control section preabsorbed using the TRPV1 blocking peptide (+BP). (B) Confocal image stack through GCL and NFL showing labeling for TRPV1 in wholemount preparation counterlabeled with antibodies against heavy-chain neurofilaments that recognize broad-field RGCs (SMI32). Image shows punctate localization to dendrites (arrows) as well as intense label to cell bodies (brackets); smaller cell bodies with TRPV1 label are in the background. (C) Confocal image stack through GCL and NFL of peripheral retina shows TRPV1 in RGC cell bodies (bracket) and in small bundles of RGC axons (arrows). (D) Confocal stack from central retina shows TRPV1 in RGC cell bodies (bracket) and in axon bundles in the NFL as they course toward the optic nerve head. Amoeboid-shaped cell bodies of microglia are apparent (ovals). (E) Confocal image in single plane at GCL/NFL border shows Iba-1–labeled microglia processes colocalizing with TRPV1, as we previously demonstrated.51 TRPV1-label RGC cell bodies (brackets) and axons (arrows) are indicated for reference. (F) Immunocytochemical labeling demonstrates strong perinuclear and dendritic localization of TRPV1 in cultured RGCs counterstained with the nuclear label DAPI. Localization to dendritic processes and neurites (dashed circles) includes node-like clusters; right: these regions are shown at higher magnification. (G, top) Western blot against TRPV1 in brain and whole retina from adult rat shows band at expected molecular weight (arrowheads; 100–113 kDa). Retina demonstrates an additional band with a slightly lower molecular weight that probably corresponds to a different glycosylation state for this antibody.79 Bottom: Control Western blot with preabsorption of TRPV1 antibody using the blocking peptide prevents detection of both bands. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer; NFL, nerve fiber layer.

Thursday, February 19, 2009

Neurotrophin Expression in Airway Smooth Muscle Cells

Dr. Y.K. Prakash and his team at Mayo Clinic College of Medicine have published a study of Neurotrophin Signaling in Airway Smooth Muscle (ASM) cells. Here they demonstrate that BDNF/TrkB signaling (but not BDNF/p75NTR) in human ASM cells regulates [Ca2+]i responses to agonist in both normal ASM, as well as with inflammation induced by TNFα. These effects suggest an important role of NT signaling in inflammation-induced changes in ASM contractility.

Y.S. Prakash, Michael A Thompson, and Christina M Pabelick. Brain Derived Neurotrophic Factor in TNF Modulation of Ca2+ in Human Airway Smooth Muscle. Am. J. Respir. Cell Mol. Biol., Feb 2009; doi:10.1165/rcmb.2008-0151OC.
... goat polyclonal anti-TrkB-Cat#: GT15080 (Neuromics, Minneapolis, MN; 1:1000 dilution)...

Featured Reagents
TrkB-Cat#: GT15080
TrkB-Cat#: MO15042
Related Reagents:
TrkB-Cat#: MO15025
TrkA for FC
TrkA Cat#: MO15034
TrkA Cat#: GT15153
All Neurotrophins and Growth Factor Antibodies
All Neurotrophins-Neuron/Glial Marker Antibodies

Thursday, February 12, 2009

Substance-P Pub

We re-introduced our Substance P Whole Serum Guinea Pig and Substance P Purified-Guinea Pig Antibodies in 2008.

The feedback has been positive. Related to this, here's a recent reference:

Maureen S. Riedl, Stephen A. Schnell, Aaron C. Overland, Anne-Julie Chabot-Doré, Anna M. Taylor, Alfredo Ribeiro-Da-Silva, Robert P. Elde, George L. Wilcox, Laura S. Stone.
Coexpression of 2A-adrenergic and -opioid receptors in substance P-containing terminals in rat dorsal horn. The Journal of Comparative Neurology Volume 513 Issue 4, Pages 385 - 398 Published Online: 29 Jan 2009 Copyright © 2009 Wiley-Liss, Inc., A Wiley Company.

...guinea pig anti-SP (1:500; Neuromics Antibodies)...

Related Reagents:

Substance P-Rabbit
Substance P-Mouse
Neurokinin-1 (NK 1) (RA25001)
Neurokinin-1 (NK 1) Human (RA25003)
Neurokinin-3 (NK 3) (RA25001)
proNeurokinin B (proNKB or P2)
All Neuropeptides

Image: Immuofluorescent detection of Substance P in rat spinal cord dorsal horn (red fluorescence). DAPI (blue) was used as counter stain.