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.