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:
Nestin
Musashi-1
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

Wednesday, September 16, 2009

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.

Description/Data:
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).


STEMEZ hNP1 FAQs


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
NT-3
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.
DOI:10.1095/biolreprod.108.075200

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
Inventors:
Stice; Steve (Athens, GA)Shin; Soojung (Baltimore, MD)Dhara; Sujoy (Athens, GA)
Assignee:
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
FACS/Phenotyping
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