Saturday, November 22, 2008

Excellent TRPV1-N IHC and WB

Kudos to Dr. Federica MF van Dissel-Emiliani and her team for the excellent Immunohistochemistry and Western Blot results using our TRPV1-N Antibody(Catalog #: RA10110) . The antibody was in their study demonstrating the sensitivity of spermatogenesis to capsaicin.

Here's the related publication:

Sefika C Mizrak, Bart M Gadella, Hatice Erdost, Aytekin Ozer, Ana MM van Pelt, Federica MF van Dissel-Emiliani. Spermatogonial stem cell sensitivity to capsaicin: An in vitro study. Reproductive Biology and Endocrinology 2008, 6:52 doi:10.1186/1477-7827-6-52.
Anti TRPV1 antibody staining: Bouin's fixed, paraffin embedded 5 um-thick rat testis sections were deparaffinized and boiled in a microwave oven (700 Watt) 3x10 min in sodium citrate buffer (0.1 mM, pH=6) for antigen retrieval. All subsequent incubations were performed for 1 hour at room temperature. The slides were then blocked with 5 % goat serum in 1 % BSA/PBS and incubated with the rabbit anti human - VR1 antibody (Neuromics, Edina, MN, USA; 1:500 in 1% BSA/PBS). Biotinilated goat anti-rabbit secondary antibody (BA-1000, Vector Labs; 1:200 in 1% BSA/PBS) was then applied. The ABC kit was finally used according to the manufacturer's instructions. Antibody reactivity was finally detected by diaminobenzidine staining (DAB, Sigma, St. Louis, MO, USA). Sections were counterstained with hematoxylin, dehydrated, mounted with Pertex and studied. Goat serum was applied on control sections.
Image: Photomicrograph of a section through an adult rat testis showing TRPV1 labelling of premeiotic germ cells, at stage II of the seminiferous epithelium. Arrow, undifferentiated spermatogonia; arrow head, early pachytene spermatocytes; asterisk, Sertoli cells.
SDS-PAGE and Western blotting: Protein lysates from the cell lines Gc-5spg and Gc-6spg and the control glioma cell line (A10-85) were prepared in RIPA buffer (PBS, 1% NP40, 0.5% sodium deoxycholate, 0.1% SDS) including 1 mM phenylmethylsulfonylfluoride. Of each sample, 50 μg were separated on a 12% SDS-polyacrylamide gel and blotted onto a polyvinylidene fluoride membrane (Millipore Corp., Bedford, MA, USA). Western blots were blocked using Blotto-A, containing 5% Protifar (Nutricia, Zoetermeer, The Netherlands) in Tris-buffered saline (10 mM Tris; 150 mM NaCl, pH 7.6), including 0.05% Tween-20. Rabbit polyclonal anti-VR1 antibody (Neuromics) was diluted 1:1000 in Blotto-A and incubated for 1 h at room temperature. Blots were washed with Tris-buffered saline with 0.05% Tween-20. After incubation with goat anti-rabbit-HRP (P-0260 Dako Cytomation, 1:5000 inBlotto-A) secondary antibody for 1 h, blots were incubated with the electrochemiluminescence kit (ECL, Amersham Pharmacia Biotech, Little Chalfont, UK)and exposed to an x-ray film (RX-omat, Kodak, Chalone / Saone, France).

Wednesday, November 12, 2008

DRG Neurons Now Available.

Primary Rat DRGs are live neurons isolated from micro-surgically dissected regions of day 18 embryonic Sprague/Dawley rat brain. These cells are prepared fresh each week and shipped in a nutrient rich medium that keeps the cells alive for up to 14 days under refrigeration.
Please note: It is important to review Protocol/Datasheet prior to ordering. There is a unique step for making the dissociation enzyme solution. Do not hesitate to call or e-mail me (612-801-1007 or should you have questions.

Image: DRGs cultured on Calf Skin Collagen.

-A dissection and Tissue Culture Manual of the Nervous System (1989). A. Shahar, J.D. Vellis, A. Vernadakis, B. Haber (Eds.), Dissociated Spinal Cord - Dorsal Root Ganglion Cultures on Plastic Tissue Culture Dishes and Glass Coverslips and Wells (pp.219-222). Wiley-Liss, Inc. J.L. Werth, -S.A. Thayer (1994) Mitrochondria Buffer Physiological Calcium Loads in Cultured Rat Dorsal Root Ganglion Neurons, The Journal of Neuroscience, 14(1), 348-356

Monday, November 03, 2008

Power Trio of Pain Research Antibodies

I received a call from a customer asking if there were any publications referencing our rabbit Alpha 2a Adrenergic Receptor antibody. The resulting search found an article referencing excellent results using a trio of of our pain research antibodies. The research was conducted by our friend, Dr. Hui-Lin Pan and his team at University of Texas M.D. Anderson Cancer Center.

Shao-Rui Chen, Hao-Min Pan, Timothy E. Richardson, and Hui-Lin Pan. Potentiation of Spinal α2-Adrenoceptor Analgesia in Rats Deficient in TRPV1- Expressing Afferent Neurons. Published online 2007 March 24. doi: 10.1016/j.neuropharm.2007.03.009.

Images: Double immunofluorescence labeling showing α2C-AR- and TRPV1-immmunoreactivity in the spinal cord dorsal horn of one vehicle-treated and one RTX-treated rat. Representative confocal images showing α2C-AR- and VR1 C-terminus (TRPV1)-immunoreactivities in the spinal dorsal horn of one vehicle- and one RTX-treated rat. All images are single confocal optical sections. Scale bar, 100 μm. Inset: high-magnification images (scale bar = 10 μm) showing the distribution of α2C-AR- and TRPV1-immunoreactivity in the lamina I and II. Neuropharmacology. 2007 June; 52(8): 1624–1630.

Rabbit VR1 N-Terminus (TRPV1) is also referenced.

Related Reagents:
Alpha 2c
Pain and Inflammation Antibodies
Vision and Retina Antibodies

Thursday, October 16, 2008

Purinergic Receptor Pubs

Our Purinergic Receptor Antibodies have recently been referenced in several publications.

In the first study 5 of our antibodies are used. It investigates the expression of purinergic P2 receptors in oxygen-induced retinal neovascularization.

Sylvia Sarman; Jorge Mancini; Ingeborg van der Ploeg; J. Oscar Croxatto; Anders Kvanta; Juan E. Gallo. Involvement of Purinergic P2 Receptors in Experimental Retinal Neovascularization. Current Eye Research. 10.1080/02713680701885470 .

...Five eyes (5 animals) from either hyperoxa- or non-hyperoxia-treated mice were enucleated and fixed for 48 hr at 4°C in paraformaldehyde (Sigma-Aldrich, St Louis, MO, USA). Afterward, eyes were immersed for cryoprotection in graded sucrose solution (5% overnight, 10%, 15%, and 20%) and interlocked with resin. Fourteen micron sections were obtained (Shandon AS325 Retraction Microtome, Thermo Scientific, Waltman, MA, USA) and fixed on polylysine-treated glass slides. After overnight incubation in primary antibody (P2X1 1:1000, P2X2 1:1000,P2X3 1:1000, and P2Y2 1:800) (Neuromics, Minneapolis, MN, USA) at 4°C, sections were treated with biotinylated secondary antibodies followed by an avidin peroxidase complex step (Vectastin Elite ABC, Vector Laboratories, Burlingame, CA, USA). Finally, a color reaction was obtained using 3,3'-diaminobenzidine (DAB)/nickel-enhanced solution for staining (Sigma-Aldrich, St Louis, MO, USA). At least 6 sections per retina were analyzed. For immunofluorescence, the sections were labeled with lissamine rhodamine-conjugated goat anti-rabbit Ig-G or with fluorescein-5-isothiocyanate-conjugated goat anti-guinea pig Ig-G (Jackson Immunoresearch Laboratories, West Grove, PA, USA). At least 6 sections per retina were analyzed. Visualization was done using a Nikon Fluorescence Eclipse Microscope (Tokyo, Japan), and photographs were taken with a Nikon DN 100 Digital Camera (Tokyo, Japan)...

Images: Expression of P2X and P2Y receptors in normal mice and in a model of ischemia-induced pathological retinal angiogenesis as assessed by immunohistochemistry. (A) P2X2 receptor expression was found in the outer plexiform layer of control mice. (B) In mice
treated with oxygen, the expression was similarly found in the outer plexiform layer. In addition, a strong signal was also found in the inner plexiform layer. This induction was noted in all slides analyzed. (C) Retina without primary antibody for P2X2. (D) In control mice,
weak P2Y2 receptor expression was seen in the ganglion cell and in the nerve fiber layer. (E) In mice treated with oxygen, P2Y2 expression
showed a similar overall distribution as control mice. The signal was, however, strongly up-regulated. This increase was noted in all slides analyzed. (F) Retina without primary antibody for P2Y2. Black bar = 20 μm.

The second studies P2Xs' role in neurotransmision.

Elsa Fabbretti*, Elena Sokolova*, Lara Masten, Marianna D’Arco, Alessandra Fabbro, Andrea Nistri and Rashid Giniatullin. Identification of negative residues in the P2X3 ATP receptor ectodomain as structural determinants for desensitization and the Ca2+ sensing modulatory sites. JBC Papers in Press. Published on October 8, 2004 as Manuscript M409772200.

...Western immunoblots of transfected or untransfected HEK cells were performed as recently reported (19). In brief, these were lysed using a buffer containing 100 mM Tris HCl (pH 6.8), 200 mM dithiothreitol, 4 % SDS, 20 % glycerol and a cocktail of protease inhibitors (Sigma), and separated on 10 % polyacrylamide gel. After blocking with Tris saline buffer (TBS) containing milk, Tween 20 and preimmune serum, proteins were incubated overnight at 4°C with an anti-P2X3 antibody (Neuromics; 1:2000). Immunocomplexes were incubated for 1 h with a peroxidase-conjugated secondary antibody (Sigma; 1:4000) and detected with a chemiluminescence ECL kit (Amersham). Negative controls were obtained by mock transfection of HEK cells with the pEGFP-N1 plasmid (CLONTECH). Controls for efficient loading of different lysate materials were carried out by using an anti-β actin antibody (mouse monoclonal, 1:2000, Sigma)...

Related Reagents:

P2X1-Rabbit Antibody
P2X2-Guinea Pig Antibody
P2X2-Rabbit Antibody
P2X3-Rabbit Antibody
P2X3-Guinea Pig Antibody
Related Reagents:
All Purinergic Receptor Antibodies
All Pain and Inflammation Antibodies
Vision and Retina Research Antibodies

Thursday, October 09, 2008

Neuron/Neuron Glial Markers

We have a comprehensive and growing catalog of Neuron/Glial Marker Antibodies and Proteins.

These reagents must work everytime in our customers' applications. Quality is confirmed by pro-active customers follow up and publication referencing the reagents.

Here we have several recent publications.

We are pleased to first feature Dr. Dr. Juana Maria Pasquini, University of and colleagues from University of Buenos Aires. She and her team use our Olig1,2,3 as a marker to study de-myelinating disease.

P.G. Franco, L. Silvestroff, E.F. Soto and J.M. Pasquini. Thyroid hormones promote differentiation of oligodendrocyte progenitor cells and improve remyelination after cuprizone-induced demyelination. doi:10.1016/j.expneurol.2008.04.039
...Olig 1-2-3 antibodies were from Neuromics Antibodies (Edina, MN); ...

Featured Product:

Related Products:
Neuron-Glial Markers
Neurotrophins and Growth Factors
Neurodegenerative Disease
Neurotrophins-Neuron/Glial Markers
Neurodegenerative Disease
SC Reagents

The second references one of our GFAP antibodies.

Sun Jin-qiao, Sha Bin, Zhou Wen-hao and Yang Yi. Basic fibroblast growth factor stimulates the proliferation and differentiation of neural stem cells in neonatal rats after ischemic brain injury. doi:10.1016/j.braindev.2008.06.005.
For the immunofluorescence assays, sections from the SVZ were washed (0.1 M Tris, pH 7.6, 15 min), denatured (2 N HCl, 37oC, 30 min), rinsed (0.1 M P10 min), incubated with 1% H2O2 in 0.1 M Tris for 30 min, rinsed, blocked (10% normal goat serum, 37oC, 30 min).
...GFAP (1:100, Neuromics)...

Saturday, October 04, 2008

siRNA-mediated gene silencing

Dr. Josephine Lai (Professor of Pharmacology, University of Arizona) is a pioneer in developing experimental designs and methods for delivering siRNA to the CNS for gene expression analysis.

She and her team have documented these in the publication:

For researchers desiring to effectively deliver siRNA to the CNS for gene expression of analysis of specific receptors, this publication offers proven methods. These include:

  • The Choice of siRNA
  • Choosing and Optimizing Transfection Reagents for siRNA Delivery to the Nervous System
  • Delivery Systems-Microinjection and Infusion (using mini-osmotic pumps)
  • Validation

We will continue to track advances by Dr. Lai and team

Wednesday, October 01, 2008

Friday, September 19, 2008

PDGFs in Non-small Cell Lung Cancer Tumor and Stromal Cells

Serving Cancer and Tumorigenesis Researhers is a central component to our business strategy. This means constantly building our catalog of Cancer Antibodies and Cancer Proteins.

I personally follow up with every customer that purchases our Cancer Research Reagents. This ensures they work as expected. In most cases, this is validate. If not, we fix the expressed issues.

We are now seeing more confirmation by the reagents being referenced in publications. In this publication, our PDGF-C antibody was used as a marker for Non-small Cell Lung Cancer Tumors.

"In univariate analyses, high tumor cell expression of PDGF-B (p  0.001), PDGF-C (p  0.01), and PDGFR- (p  0.026) were negative prognostic indicators for disease-specific survival."

Donnem, Tom MD, Al-Saad, Samer MD, Al-Shibli, Khalid MD, Andersen, Sigve MD, Busund, Lill-Tove MD, PhD, Bremnes, Roy M. MD, PhD. Prognostic Impact of Platelet-Derived Growth Factors in Non-small Cell Lung Cancer Tumor and Stromal Cells. Journal of Thoracic Oncology. 3(9):963-970, September 2008.

Featured Reagent:

Related Reagents to Consider:
PDGF R Alpha
Cancer Antibodies
All Neurotrophin and Growth Factor Antibodies
Platelet Derived Growth Factor Proteins
Cancer Research Proteins
Neurotrophin and Growth Factor Proteins

Thursday, September 18, 2008

Rat Sensory Neurons and NK-1

A publication featuring our Neurokinin-1 (NK 1) Receptor just crossed my radar. It is authored by our friend, Dr. Matt Ramer.

Matt and his team at University of British Columbia study primary sensory nerve cells (neurons), which are responsible for the transmission of somatic (bodily) sensations such as touch, pain, hot, cold and so on from the periphery (skin, muscles and viscera) to the central nervous system (CNS, spinal cord and brain). His research extends to therapeutic potential of neurotrophins on regeneration in spinal cord injury and deafferentation pain.

In the past several years, he has generouly shared NT-3 and BDNF IHC data with us.

Here's a link to the publication:

Matt Ramer. Anatomical and functional characterization of neuropil in the gracile fasciculus. The Journal of Comparative Neurology. 10.1002/cne.21785.

Featured Reagent
Neurokinin-1 (NK 1) Receptor

Other Reagents to Consider:
Neurokinin-1 (NK 1) Human (RA25003)
Neurokinin-3 (NK 3) (RA25002)
Substance P (GP14103)
proNeurokinin B (RA25008)
All Neuropeptides
All Pain and Inflammation

Saturday, September 13, 2008

ELISA techniques and protocols

Power Point of ELISA Methods and Protocols(pps - 1.4M)
This teaching guide covers the three major types of ELISA: indirect, competitive, and sandwich. It integrates theory with practice, to help you understand what you are doing, and help you to do it!

Courtesy of our friends at Novus Biologicals.

Thursday, September 11, 2008

Ischemia, Inflammatory Response and Umbilical Stem Cells

We would like to send Kudos to Dr. Yan Xu and his colleagues at University of Pittburgh for their findings on inflammatory response in Golbal Ischemia. Their work was recently published:

Aaron Hirko, Renee Dallasen, Sachiko Jomura, Yan Xu. Modulation of Inflammatory Responses after Global Ischemia by Transplanted Umbilical-Cord Matrix Stem Cells. Stem Cells First published online August 21, 2008; doi:doi:10.1634/stemcells.2008-0075

Secondary to Cardiac Arrest is Brain Damage do to lack of blood flow. This is marked by a delayed loss of Neurons in CA1 hippocampus region of the brain due to inflammatory response.

The story timeline of this response is good then bad with interesting twists. The delay in neuronal loss is linked to initial inflammation. It involves both reactive astrocytes (astrocytosis) and glia. Delaying the loss is, of course, good.

...But then, the reactive astrocytosis and related glial scarring cause a physical and biochemical barrier to regeneration of neurons...a bad thing. Protecting the microglia is a good thing, because they these cells serve as scavengers for clearing the cellular debris. They can also secrete a variety of cytotoxic and protective chemicals.

The wow factor in this research is that implanted rat umbilical-cord matrix (RUCM) cells can provide partial protection against neuronal injury in rat brains. Rats treated with RUCM cells three days prior to an 8-min CA had only 25-32% neuronal loss in the hippocampal CA1 region compared to the typical 50-68% neuronal loss observed in the untreated or the vehicle-treated animals. This could be due to to the favaorable modulation of the "good-bad" inflammatory response.

The good news in the search for therapies for stroke and cardiac arrest victims is combined, stem-cell-like RUCM cells offer protection against neuronal injury after global cerebral ischemia by enhancing the survivability of the astroglia in the selectively vulnerable regions.

We are pleased that the research team used our GFAP antibody as an marker for astrotytic in their studies.

Monday, September 08, 2008

Neurotoxicity Testing

We combine our expertise in providing fresh and healthy:

E18 and E20 Rat Primary Neuronal Tissue -NEURON CULTURES

E18 Rat Primary Neuronal Tissue - ASTROCYTE CULTURES

E18 Mouse Neuronal Tissue -NEURON CULTURES

E18 Mouse Neuronal Tissue -ASTROCYTE CULTURES

AND Apoptosis Research Reagents

to help researchers more effectively study Neurotoxicity.

Images: Polycaspase Assay Kit, green was used to assess cell death in primay rat hippocampal neurons.Cells were plated on 25-mm poly-l-lysine-coated coverslips at 300,000 cells per coverslip. Cells were used at 4 or 8 days in vitro. Composite imagae (A) 3 out of 4 cells are apoptotic (green). No cells were necrotic as both of the PI-positive cells were FLICA-positive; they had compromised membranes and were probably in the late stages of apoptosis rather than necrosis. (B) 3 Caspase-positive cells fluoresce green.

Thursday, August 28, 2008

No Pain; No Gain

We work hard to make sure our Pain and Inflammation antibodies continue to be a gold standard for researchers. We follow up with virtually all researchers to make sure they work to expecations in each unique application.

We also look for references in current publication. Although published in 2006, this one just crossed our radar scope.
It contains multiple images of 3 of our top sellers: Mu Opioid Receptor, VR1 C-terminus (TRPV1) and VR1 N-Terminus (TRPV1).

Monday, August 18, 2008

Transthyretin, αAPP peptides and Alzheimer's Disease

Demographics point to increasing rates of Alhzheimer's Disease. This disease steals away the golden years of sufferers. It also costs society billions healthcare.

Research for the cure marches on. Researchers know the disease is characterizedby the deposition of amyloid β-peptide (A-Beta) in the brain. The challenge is finding a way to protect the brain from these depositions or reverse the process.

Dr. Isabel Cardoso and her team at Instituto de Biologia Molecular e Celular have recently published compelling work in this area. Here they provide more important evidence for the role of a Transthyretin (TTR) protective mechanism. This mechanism could include the removal of deposited A-Beta.

A-Beta proteolysis by TTR is KPI-sensitive.
A- A-Beta incubated with TTR (A-Beta+TTR) shows a weaker A-Beta monomer band as compared to A-Beta alone (A-Beta), indicative of proteolysis, as analyzed by SDS-PAGE electrophoresis followed by western blot. Pre-incubation of TTR with pefabloc (A-Beta+(TTR+pefabloc)) and with an αAPP peptide containing the KPI domain (A-Beta+(TTR+KPI+−APP)) inhibits TTR proteolytic activity, whereas the αAPP peptide without the KPI domain (A-Beta+(TTR+KPI−−APP)) facilitates proteolysis. B- % of inhibition of TTR proteolysis by quantification of band intensity in A. C- Ultrastructural analysis by TEM of preparations incubated for 15 hours, as described in Materials and Methods. TTR inhibited A-Beta aggregation as compared with A-Beta incubated alone (upper panels). Pre-incubation of TTR with αAPP peptide containing the KPI domain (A-Beta+(TTR+KPI+−APP)) abrogated TTR ability to avoid A-Beta aggregation, whereas αAPP lacking the KPI domain (A-Beta+(TTR+KPI−−APP)) did not affected TTR activity (lower panels). Scale bar=500 nm.

Neuromics' Reagent Used-APP 228

MMP-9, E. Coli and Breast Cancer

The application of live bacteria for cancer therapies holds promise. In this study, Escherichia coli K-12 colonization on the tumour microenvironment by immunohistochemistry and fluorescence microscopy in the murine 4T1 breast carcinoma model. MMP-9 and TNF-alpha expression is altered in the sites of tumors.

Interestingly, the authors observed a postive change in the expression of these proteins after colonization.

Stephanie Weibel, Jochen Stritzker, Matthias Eck, Werner Goebel, and Aladar A. Szalay. Colonization of experimental murine breast tumours by Escherichia coli K-12 significantly alters the tumour microenvironment. 10.1111/j.1462-5822.2008.01122.x. Cellular Microbiology. © 2008 Blackwell Publishing Ltd
...goat anti-mouse MMP-9 antibody (Neuromics, Edina, MN)...

Monday, August 11, 2008

Cholesterol Homeostasis and Diabetes

Hormone-sensitive lipase (HSL) is widely expressed in adipose tissue. Interestingly, HSL-null mice have been shown to be resistant to diet-induced obesity. Despite this characteristic, they can also show insulin resistance. This resistance is contributing factor in type 2 diabetes.

Scientists have also shown that pancreatic beta cells, responsible for insulin release, begin to malfunction when their cholesterol levels build up. So what is the link?

The study referenced here suggests that HSL plays a critical role in the hydrolysis of cytosolic cholesteryl esters and that increased levels of hepatic cholesteryl esters, due to lack of action of HSL in the liver, is the main mechanism underlying the imbalance in cholesterol metabolism in HSL-null mice.

Celine Fernandez, Marie Lindholm, Morten Krogh, Stéphanie Lucas, Sara Larsson, Peter Osmark, Karin Berger, Jan Boren, Barbara A Fielding, Keith N. Frayn and Cecilia Holm. Disturbed cholesterol homeostasis in hormone-sensitive lipase null mice. Am J Physiol Endocrinol Metab (July 29, 2008). doi:10.1152/ajpendo.90206.2008.

...Liver samples were homogenized in lysis buffer pH 7.0 containing 25 mM Tris, 150 mM NaCl, 1 mM EDTA, 1% Triton and 1X protease inhibitor cocktail (Complete Mini, Roche). Total protein concentration was measured by BCA assay (Pierce) and 50 μg of protein were subjected to SDS-polyacrylamide gel electrophoresis (8 % polyacrylamide). After transfer to PVDF membranes (Invitrogen), blots were incubated with a primary antibody mouse anti-mouse/rat ABCA1 (Neuromics) according to the instructionsof the manufacturer. As secondary antibody a horseradish peroxidase-conjugated sheep antimouse IgG was used. Western blot analysis was performed using a chemiluminescence system (Luminol) and detection was made using a CCD-camera (LAS 1000, Fuji). Band intensities were quantified using the ImageJ software (

MMP-9 and Neuroinflammation & Autoimmunity

Zhi-Yuan Zhang, Zhiren Zhang, Uwe Fauser and Hermann J. Schluesener. Improved outcome of EAN, an animal model of GBS, through amelioration of peripheral and central inflammation by minocycline. DOI. 10.1111/j.1582-4934.2008.00333. © 2008 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.
... Metalloproteinase 9 (MMP-9); 1:500; Neuromics, Edina, MN, USA...

Related Reagents:

proMMP-7 (Human)
MMP-24 (MT5-MMP, Human)
Cancer Antibodies
Neurodegenerative Disease Antibodies
Pain and Inflammation

Thursday, August 07, 2008

New Neurotensin Receptor (NTS-1) antibody

Our customers have had success with these antibodies.

Here's a representative publication:

We are pleased to announce the release of a chicken Neurotensin Receptor 1 (NTS1).
Image: Immuofluorescent detection of NTS1 in cell bodies of neurons in rat dorsal root ganglia fluorescence).

Tuesday, August 05, 2008

TRPV1 Staining of Mouse DRG

This excellent staining comes to us courtesy of Katharina Zimmermann (Childrens Hospital Boston, Clapham Lab). This is some of the best staining we've seen using our VR1 C-Terminus (TRPV1) - mouse specific antibody.

Images: TRPV1 staining of C57BL/6 mouse dorrsal root ganglia. Tissues were stained using Alexa Fluor© 488 (Green) and counterstained with DAPI (blue).

cryosections, 10 microns thickness

Friday, August 01, 2008

Stressed Induced Analgesia (SIA) and OFQ

This is to highlight excellent work done by our friends at SRI and AfaSci.

Our Orphanin FQ ab was used in experiments by the team. Here's the latest publication highlighting their work:

Xinmin Xie, Jonathan P. Wisor, Junko Hara, Tara L. Crowder1, Robin LeWinter, Taline V. Khroyan, Akihiro Yamanaka, Sabrina Diano, Tamas L. Horvath, Takeshi Sakurai, Lawrence Toll and Thomas S. Kilduff. Hypocretin/orexin and nociceptin/orphanin FQ coordinately regulate analgesia in a mouse model of stress-induced analgesia. J. Clin. Invest. 118(7): 2471-2481 (2008). doi:10.1172/JCI35115. Copyright © 2008, The American Society for Clinical Investigation.

...Brain sections (10 μm) were treated with 0.3% H2O2 to quench endogenous peroxidases and then incubated overnight in primary anti-N/OFQ (1:5,000; RA10106, anti-FGGFTGARKSARKLANQ, Neuromics) and anti-orexin-B (1:5,000; sc-8071, Santa Cruz Biotechnology Inc.) antisera at 4°C with agitation. Sections were incubated in blocking buffer for 1 hour, followed by a 2-hour incubation at room temperature in secondary antisera (Alexa Fluor 546 donkey anti-goat [1:750] and 488 donkey anti-rabbit [1:500]; Molecular Probes, Invitrogen). As a negative control, additional sections were treated similarly, but the primary antibody was omitted. Preadsorption with the N/OFQ peptide FGGFTGARKSARKLANQ was used as a positive control and blocked all specific staining as also found by others...

N/OFQ-containing fibers innervate Hcrt neurons, and N/OFQ inhibits Hcrt neuronal activity. (A) Left panel shows confocal image of N/OFQ-immunoreactive fibers in the vicinity of, and in putative contact with, Hcrt-immunoreactive neurons in the PLH of WT mice. N/OFQ (green) fibers are in close proximity to Hcrt-immunoreactive (red) cells. The arrow indicates the N/OFQ innervation of an Hcrt cell body. Middle panel shows light micrograph of a light brown immunolabeled Hcrt neuron contacted by a dark black bouton (arrow) representing immunolabeling for N/OFQ. Right panel shows electron micrograph taken from ultrathin sections of the same labeled terminal and dendrite shown in the light micrograph in the middle panel. Black arrow indicates the N/OFQ-immunolabeled axon terminal in synaptic contact (red arrowhead) with the dendrite of the Hcrt cell. Scale bars: 10 μm (left and middle panels); 1 μm (right panel). (B) Under current clamp, bath application of N/OFQ (1 μM) hyperpolarized Hcrt neurons, decreased input resistance, and blocked spontaneous firing of action potentials. The resting potential of this cell was –54 mV and was manually adjusted to –60 mV with DC current. Membrane resistance was monitored using hyperpolarizing current pulses (–0.3 nA, 800 ms) delivered every 5 seconds throughout the experiment. (C) Under voltage-clamp mode at a Vh of –60 mV, N/OFQ (1 μM) induced an outward current (–53 pA) in an Hcrt neuron. Notice that the frequency but not the amplitude of the miniature synaptic currents (inward currents) recorded in the presence of TTX (0.5 μM) was also reduced.

Wednesday, July 30, 2008

TH-Marker for Diabetic Neuropathy

New Pub Referencing our Tyrosine Hydroxylase.

Blount, Andrew L. B.A.; Peled, Ziv M. M.D.; Dexter, Erica L. B.S.; Nagle, Raymond B. M.D., Ph.D.; Maloney, Christopher T. M.D.; Dellon, A Lee M.D., Ph.D. Sympathetic Nerves in the Tarsal Tunnel: Implications for Blood Flow in the Diabetic Foot. Plastic & Reconstructive Surgery. 122(1):188-191, July 2008.

...Specimens were fixed in 10% neutral buffered formalin for 3 to 4 hours and then embedded in paraffin. Three-micron sections were obtained from the block and used for immunohistochemistry, which was performed using a mouse monoclonal antibody against tyrosine hydroxylase (MO20001; Neuromics, Edina, Minn.) diluted 1:50 in Discovery Diluent (Ventana Medical Systems, Inc., Tucson, Ariz.) on a Discovery XT Automated Immunostainer (Ventana Medical Systems)...
Images: (A) Tyrosine hydroxylase immunohistochemical staining of tibial epineurium from the tarsal tunnel. Positively staining sympathetic fibers (arrows) are seen within a larger nerve bundle. (B) Tyrosine hydroxylase immunohistochemical staining of connective tissue from the tarsal tunnel. Several positively staining sympathetic fibers (arrows) are seen innervating the media of a local venule.

Thursday, July 24, 2008

SP is Back and Better than Ever

We have successfully re-made our widely used Guinea Pig Substance P antibody. It has been tested both internally and by an interested customer. The results exceeded expectations.

This antibody has proven useful for Pain/Inflammation and Diabete/Obesity Research.

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

Here're are related publications:

P . Tsai , A . Alonso , M . Pinto , D . Leigh , E . Weiler , J . Fricton , M . Erickson , L . Stone , L . Kehl. Substance P is co-localized with protein gene product 9.5-immunoreactive nerve fibers in intervertebral discs from patients with painful degenerative disc disease. doi:10.1016/j.jpain.2006.01.096
...guinea pig anti-substance P (1:500; Neuromics, Edina, MN) ...
Mei Bigliardi-Qi, Claire Gaveriaux-Ruff, Katrin Pfaltz, Pierre Bady, Tommy Bauman, Theo Rufli, Brigitte L Kieffer and Paul L Bigliardi. Deletion of - and -Opioid Receptors in Mice Changes Epidermal Hypertrophy, Density of Peripheral Nerve Endings, and Itch Behavior. Journal of Investigative Dermatology (2007) 127, 1479–1488. doi:10.1038/sj.jid.5700661; published online 21 December 2006.
.. substance P (Neuromics 1:200) staining, the sections were incubated at 4°C over night in Zamboni buffer. ...

Related Reagents:
Neurokinin-1 (NK 1) Receptor
Neurokinin-1 (NK 1) Human Receptor
Neurokinin-3 (NK 3) Receptor
proNeurokinin B (proNKB or P2)
All Neuropeptides
Pain and Inflammation Antibodies
Diabetes and Obesity Antibodies
Substance P Customer Data

Wednesday, July 23, 2008


We will be releasing soon a website dedicated to ELISA and ELISpot.

Here are excellent "how to" videos.

Sunday, July 13, 2008

Molecular Pathways and Heart Development

Owen WJ Prall, Mary K Menon, Mark J Solloway, Yusuke Watanabe, Stéphane Zaffran, Fanny Bajolle, Christine Biben, Jim J McBride, Bronwyn R Robertson, Hervé Chaulet, Fiona A Stennard, Natalie Wise, Daniel Schaft, Orit Wolstein, Milena B Furtado, Hidetaka Shiratori,6 Kenneth R Chien, Hiroshi Hamada,6 Brian L Black, Yumiko Saga, Elizabeth J Robertson, Margaret E Buckingham, and Richard P Harvey. An Nkx2-5/Bmp2/Smad1 negative feedback loop controls second heart field progenitor specification and proliferation. Cell. 2007 March 9; 128(5): 947–959. doi: 10.1016/j.cell.2007.01.042.

Summary: During heart development the second heart field (SHF) provides progenitor cells for most cardiomyocytes and expresses the homeodomain factor Nkx2-5. We now show that feedback repression of Bmp2/Smad1 signaling by Nkx2-5 critically regulates SHF proliferation and outflow tract (OFT) morphology. In the cardiac fields of Nkx2-5 mutants, genes controlling cardiac specification (including Bmp2) and maintenance of the progenitor state were up-regulated, leading initially to progenitor over-specification, but subsequently to failed SHF proliferation and OFT truncation. In Smad1 mutants, SHF proliferation and deployment to the OFT were increased, while Smad1 deletion in Nkx2-5 mutants rescued SHF proliferation and OFT development. In Nkx2-5 hypomorphic mice, which recapitulate human congenital heart disease (CHD), OFT anomalies were also rescued by Smad1 deletion. Our findings demonstrate that Nkx2-5 orchestrates the transition between periods of cardiac induction, progenitor proliferation and OFT morphogenesis via a Smad1-dependent negative feedback loop, which may be a frequent molecular target in CHD.

...goat anti-Isl1, raised against full length human Isl1, GT15051, Neuromics)...

Monday, June 30, 2008

Combined Neuron Cultures Assays

We would like to thank Dr. Matteo Porotto, Weill Cornell Medical College

Larger Image

Primary rat neurons: Transfection of functional HeV glycoproteins and infection with HeV pseudotyped virions.In order to establish the feasibility of carrying out the proposed experiments in primary neurons, we show (figure ) that our assays are amenable to use in primary neurons. In the experiment, Combined Hippocampus, Cortex, and Ventricular -E18 (Neuromics) were plated, and at 3 days were transfected with plasmids encoding HeV G/F as well as YFP. On the following day, these cells were infected with HeV or VSV pseudotyped viruses bearing RFP. In the figure, (A) the phase contrast photos show the differentiated neurons; (B) upon excitation for RFP, the red fluorescence indicates neurons infected by HeV pseudotyped virions; (C) upon excitation for YFP, and the green fluorescence shows the efficiency of transfection in neurons. This experiment indicates that the proposed experiments can be carried out in primary neurons, which are transfectable and infectable in our systems, and thus supports all the proposed aims. Data Courtesy of Dr. Matteo Porotto, Weill Cornell Medical College

Primary Neuron Cultures Featured
Combined Hippocampus, Cortex, and Ventricular -E18
Other Reagents to Consider
All Primary Neurons/Astrocytes, Stem Cells and Media
Transfection Reagents
Neuron-Glial Markers
Stem Cell Markers

Thursday, June 19, 2008

Amped Up ORL-1

We have been collaborating with a customer to amp up our ORL-1. Here's an example of a resulting IHC images.

Image: ORL 1 staining in rat brain (singulate cortex). ORL1 Detection was done using anti-rabbit Cy3 conjugated antibodies (red color). DAPI was used to counterstain cell nuclei (blue color). Working dilution is 1:40-1:100.

Wednesday, June 04, 2008


Katrin Schnizler, Leonid P. Shutov, Michael J. Van Kanegan, Michelle A. Merrill, Blake Nichols, G. Stanley McKnight, Stefan Strack, Johannes W. Hell, and Yuriy M. Usachev. Protein Kinase A Anchoring via AKAP150 Is Essential for TRPV1 Modulation by Forskolin and Prostaglandin E2 in Mouse Sensory Neurons. J. Neurosci., May 2008; 28: 4904 - 4917 ; doi:10.1523/JNEUROSCI.0233-08.2008

......anti-TRPV1 antibody (catalog #RA14113; Neuromics, Edina, MN) and 30 mul of prewashed NaCl). Probing was done using the Neuromics anti-TRPV1 antibody (1:500 dilution...with other TRPV1 antibodies either from Neuromics (N-terminus rabbit anti-TRPV1) or from......

Thursday, April 03, 2008

Recent Pubs Referencing Neuromic's Products

Neurotensin Receptor 1 (NTS1)-GP14020

Geneviève Roussy, Marc-André Dansereau, Louis Doré-Savard, Karine Belleville, Nicolas Beaudet, Elliott Richelson and Philippe Sarret. Spinal NTS1 receptors regulate nociceptive signaling in a rat formalin tonic pain model. doi:10.1111/j.1471-4159.2007.05205.x

Images: Expression of NTS1 receptors in sensory neurons and spinal cord. (a) Immunoperoxidase staining reveals the presence of NTS1-like immunoreactivity throughout the lumbar dorsal horn of the spinal cord. Immunolabeling is most prominent in the superficial layers of the dorsal horn. The lateral spinal nucleus (lsn) is also moderately labeled (arrow). (b) High magnification of (a). A dense NTS1 immunostaining is observed over laminae I and II of the dorsal horn. Numerous immunopositive nerve cell bodies are visible in the superficial laminae and in the nucleus proprius of the dorsal horn (arrows). (c) Light microscopic analysis of NTS1 expression in primary afferent neurons. NTS1 is expressed in subpopulations of small- and medium-ganglion cells. No apparent labeling is detected in large DRG neurons (arrows). At higher magnification, NTS1 neurons exhibit a cytoplasmic pattern of immunoreactivity (d, arrowheads). (e) Identification of endogenously expressed NTS1 receptors, by western blotting, in homogenates from DRGs, lumbar spinal cord and brain. The 47 kDa protein band corresponds to the molecular weight deduced from the cDNA sequence of NTS1. Each lane represents the transfer of 25 lg of protein. Scale bars: (a, b, c, and d) 300, 150, 70 and 20 lm, respectively. doi:10.1111/j.1471-4159.2007.05205.x

Primary Neurons and Astrocytes

E18 and E20 Rat Primary Neuronal Tissue -NEURON CULTURES
E18 Rat Primary Neuronal Tissue - ASTROCYTE CULTURES
E18 Mouse Neuronal Tissue -NEURON CULTURES
E18 Mouse Neuronal Tissue -ASTROCYTE CULTURES
Frozen Primary Rat Neurons-NEW
Frozen Primary Mouse Neurons and Astrocytes-New
FluoGreen Tracer-Stains Cell Nuclei and Cytoplasm

Primary Neurons and Astrocytes

Shouqing Luo, Haruo Mizuta and David C. Rubinsztein. p21-activated kinase 1 promotes soluble mutant huntingtin self-interaction and enhances toxicity. Human Molecular Genetics 2008 17(6):895-905; doi:10.1093/hmg/ddm362

...Rat E18 cortical and striatal neurons were purchased from Neuromics (MN) or prepared from E18 rat brain tissues according to instructions from Neuromics, cultured in neurobasal (Invitrogen) medium supplemented with B27 (Invitrogen) and 0.5 mM glutamine and transfected with Lipofectamine 2000 according to standard methods...

Friday, March 07, 2008

New Data and Pubs referencing Neuromics

Cullen L. Schmid, Kirsten M. Raehal and Laura M. Bohn. Agonist-directed signaling of the serotonin 2A receptor depends on ?;-arrestin-2 interactions in vivo. Published online on January 14, 2008, 10.1073/pnas.0708862105
...Neuromics, Edina, MN; rabbit anti-5HT ..... 5A-5D...

Thomas W. Gould, Shigenobu Yonemura, Ronald W. Oppenheim, Shiho Ohmori, and Hideki Enomoto. The Neurotrophic Effects of Glial Cell Line-Derived Neurotrophic Factor on Spinal Motoneurons Are Restricted to Fusimotor Subtypes. The Journal of Neuroscience, February 27, 2008, 28(9):2131! -2146
...mouse Ret (Neuromics, Edina, MN), human Ret (Neuromics), and activated caspase-3 (Cell Signaling, Danvers, MA). Muscles or sections were rinsed and then...

Image: Nuclei of D3 or IGnT-KO ES cells stained with goat anti-Oct3/4 antibody. doi: 10.1093/glycob/cwm138

Thursday, January 17, 2008

Nov-Dec Publication Referencing Neuromics

Diversity in the Neural Circuitry of Cold Sensing Revealed by Genetic Axonal Labeling of Transient Receptor Potential Melastatin 8 Neurons
CELLULAR/MOLECULAR:Yoshio Takashima, Richard L. Daniels, Wendy Knowlton, James Teng, Emily R. Liman, and David D. McKemyDiversity in the Neural Circuitry of Cold Sensing Revealed by Genetic Axonal Labeling of Transient Receptor Potential Melastatin 8 NeuronsJ. Neurosci., Dec 2007; 27: 14147 - 14157 ; doi:10.1523/JNEUROSCI.4578-07.2007 ......211; Santa Cruz Biotechnology, Santa Cruz, CA), 1:500 rabbit anti-transient receptor potential vanilloid 1 (TRPV1; RA14113; Neuromics, Edina, MN), and 1:500 rabbit anti-TRPM8 (a gift from M. Tominaga, Okazaki Institute for Integrative Bioscience, Okazaki......
Full Text
Supplemental Data
G 5 Is Required for Normal Light Responses and Morphology of Retinal ON-Bipolar Cells
BRIEF COMMUNICATIONS:Anjali Rao, Rebecca Dallman, Scott Henderson, and Ching-Kang ChenGβ5 Is Required for Normal Light Responses and Morphology of Retinal ON-Bipolar CellsJ. Neurosci., Dec 2007; 27: 14199 - 14204 ; doi:10.1523/JNEUROSCI.4934-07.2007 ......2003) 1:100 RGS6 Imgenex (San Diego, CA) 1:100 Goalpha Millipore (Billerica, MA) 1:100 mGluR6 Neuromics (Northfield, MN) 1:100 PKCalpha Sigma 1:100 FLAG Sigma 1:100 Transmission electron microscopy......
Full Text
Exogenous Delivery of Heat Shock Protein 70 Increases Lifespan in a Mouse Model of Amyotrophic Lateral Sclerosis
NEUROBIOLOGY OF DISEASE:David J. Gifondorwa, Mac B. Robinson, Crystal D. Hayes, Anna R. Taylor, David M. Prevette, Ronald W. Oppenheim, James Caress, and Carolanne E. MilliganExogenous Delivery of Heat Shock Protein 70 Increases Lifespan in a Mouse Model of Amyotrophic Lateral SclerosisJ. Neurosci., Nov 2007; 27: 13173 - 13180 ; doi:10.1523/JNEUROSCI.4057-07.2007 ......overnight, at 4C. The primary antibodies used for glial cell activation were glial fibrillary acidic protein (GFAP; RA22101; Neuromics, Minneapolis, MN) and ionized calcium binding adaptor molecule 1 (Iba1; 016-20001; Wako, Neuss, Germany). The primary antibody......
Full Text
Modulation of Semaphorin3A Activity by p75 Neurotrophin Receptor Influences Peripheral Axon Patterning
DEVELOPMENT/PLASTICITY/REPAIR:Ayal Ben-Zvi, Liat Ben-Gigi, Hagit Klein, and Oded BeharModulation of Semaphorin3A Activity by p75 Neurotrophin Receptor Influences Peripheral Axon PatterningJ. Neurosci., Nov 2007; 27: 13000 - 13011 ; doi:10.1523/JNEUROSCI.3373-07.2007 ......domain) used in Western blot analyses was purchased from Alamone Labs (Jerusalem, Israel). Anti-Neuropilin1 was purchased from Neuromics (Edina, MN). Secondary antibodies were obtained from Jackson Immunoresearch Laboratories (Jackson, PA). Protease inhibitor......
Full Text
Supplemental Data
Somatostatin Peptides Produce Multiple Effects on Gating Properties of Native Cone Photoreceptor cGMP-Gated Channels That Depend on Circadian Phase and Previous Illumination
CELLULAR/MOLECULAR:Shih-Kuo Chen, Gladys Y.-P. Ko, and Stuart E. DryerSomatostatin Peptides Produce Multiple Effects on Gating Properties of Native Cone Photoreceptor cGMP-Gated Channels That Depend on Circadian Phase and Previous IlluminationJ. Neurosci., Nov 2007; 27: 12168 - 12175 ; doi:10.1523/JNEUROSCI.3541-07.2007 ......into the plasma membrane, and this can be monitored to infer activation of PLC. Transfection was performed using pn-Fect from Neuromics (Edina, MN). Briefly, retinal cells were grown for 4 d in LD cycles, until 24 h before analysis. At that time, coverslips......