Tuesday, May 30, 2017

Rose Hip and Heart Disease

Helps Prevent Atherosclerotic Plaques

This study features the use of our ABCA1 Antibody. The ATP-binding cassette transporter (ABC) Protein called ABCA1 has a major impact on cellular and whole body cholesterol metabolism and is likely to play an important role in protecting against cardiovascular disease.

The authors conclude that rose hip is a suitable dietary supplement for preventing atherosclerotic plaques formation by modulating systemic blood pressure and the expression of RCT and inflammatory genes.Michele Cavalera, Ulrika Axling, Catarina Rippe, Karl Swärd, and Cecilia Holm. Dietary rose hip exerts anti-atherosclerotic effects and increases nitric oxide-mediated dilation in ApoE-null mice. The Journal of Nutritional Biochemistry. Available online 21 March 2017. http://dx.doi.org/10.1016/j.jnutbio.2017.02.017...Western blotting. Approximately 50 mg of tissue was homogenized in lysis buffer (50 mM Tris–HCl, pH 7.5, 1 mM EGTA, 1 mM EDTA, 1% NP40, 1 mM Na-orthovanadate, 40 mM NaF, 4 mM Na-pyrophosphate, 0.27 M sucrose, 1 mM DTT, 20 μg/ml leupeptin, 10 μg/ml antipain and 1 μg/ml pepstatin) and subsequently centrifuged at 12,000 g for 20 min at 4 °C. Protein concentration of the supernatant was determined using BCA-assay (Pierce) and 20 μg of proteins were resolved on NuPAGE 4–12% Bis-Tris Gel (Life Technologies – Carlsbad, CA) and electroblotted onto nitrocellulose membranes (Amersham, GE Healthcare – UK). The membranes were washed and incubated with primary and appropriate secondary antibodies. Primary antibodies used were: SR-B1 (ab3 - Abcam), beta-actin (A5441 – Sigma), Abca1 (MO13101 - Neuromics), Abcg1 (orb214917, Biorbyt), Abcg5 (orb5656, Biorbyt), Abcg8 (orb228808, Biorbyt), IL1b (orb101745, Biorbyt), IL6 (orb228222, Biorbyt), NF-kB (ab7970, Abcam) and HSP90 (BD610418, BD bioscience)...

RH supplementation modulates the reverse cholesterol transport pathway. (a) Liver qPCR analysis of cholesterol metabolism showed similar expression of genes involved in the synthesis and conversion of cholesterol into bile acids but increased expression of various RCT genes upon rose hip feeding (n=5–6).
(b) Protein levels of SR-B1, Abcg1 and Abcg5 were increased in the liver of RH-feeding mice (n=6). *P<.05, **P<.01 and ^P=.056 versus CTR.

We have an extensive catalog of Characterized, Tested and Research Ready Antibodies/Markers. Check us out.

Wednesday, May 24, 2017

Parkinson's Disease and the Striatum

Excellent Video
Researchers at the Karolinska Institute recently released a video outlining their findings regarding the root causes of Parkinson's.


Striatal sensory responses were studied by whole-cell recordings and optogenetics
Dopamine (DA) depletion affects intrinsic and sensory properties in direct pathway neurons
The encoding of bilateral tactile stimuli is impaired following DA depletion
Administration of L-DOPA can correct sensory deficits caused by DA depletion.
for more see: https://doi.org/10.1016/j.neuron.2017.05.004
This study came to my attention the researchers used our Enkaphalin Antibody as a marker for the Dopaminergic Neurons. 
Image: Mouse striatum stained with D2 cell marker Enkephalin (RA14124) in green and with neuronal marker NeuN in red courtesy of Dr Heike Rebholz of City College of New York

Friday, May 19, 2017

TRPV1 Channels and Neuropathic Pain

Neuromics' TRPV1 Stain Neurons and Microglia for Study

TRPV1 is mainly functional in the microglia. Its activation, beyond controlling microglia reaction per se, modulated microglia-neuron communication, by promoting release of extracellular vesicles (EVs) from microglia. Indeed, EVs are important mediators of intercellular communication between microglia and brain cells: Maria Cristina Marrone, Annunziato Morabito, Michela Giustizieri, Valerio Chiurchiù, Alessandro Leuti, Marzia Mattioli, Sara Marinelli, Loredana Riganti, Marta Lombardi, Emanuele Murana, Antonio Totaro, Daniele Piomelli, Davide Ragozzino, Sergio Oddi, Mauro Maccarrone, Claudia Verderio & Silvia Marinelli. TRPV1 channels are critical brain inflammation detectors and neuropathic pain biomarkers in mice. Nature Communications 8, Article number: 15292 (2017) doi:10.1038/ncomms15292.

(a–f) Sections of cortical tissue from WT and TRPV1−/− mice, fixed after exposure to ACSF (a,d), ACSF plus vehicle (DMSO; b,e) and ACSF plus 1μM capsaicin (c,f) and immune-processed for iba-1 to stain microglia cells (in red). INSETs are zoom images taken from an area delimited by the yellow square for each condition. (g), Bar graph of percentage of cortical microglia cell phenotype (resting, ameboid, bushy and hypertrophied), in control (grey bars), vehicle- (dark grey bars) and capsaicin- treated (red bars) cortical sections from WT mice. Capsaicin treatment causes a significant shift from ramified and bushy to hypertrophied morphology. (h), Same as in ‘g’ but in cortical sections from TRPV1−/− mice. In these tissues capsaicin fails to induce morphological changes of microglia cells. Note that microglia cells in −/− tissues are already hypertrophied in control conditions (d–f,h).
Our Pain and Inflammation Research Antibodies Continue to be widely used and frequently published.

Monday, May 15, 2017

TRPV1 Antibodies

Designed for Your Success.
The roots of our TRPV1 Antibodies run deep. They have been key to our ongoing success. We have been providing them to Researchers since the inception of Neuromics (12 years ago).

We measure our success one Researcher at a time. Positive feedback includes references in many publications. There are indeed "Tested; Characterized and Research Ready"

Here's a pub hot off the presses: Noémi Bohonyi, Krisztina Pohóczky, Bálint Szalontai, Anikó Perkecz, Krisztina Kovács, Béla Kajtár, Lajos Orbán, Tamás Varga, Sarolta Szegedi, József Bódis, Zsuzsanna Helyes, Miklós Koppán. Local upregulation of transient receptor potential ankyrin 1 and transient receptor potential vanilloid 1 ion channels in rectosigmoid deep infiltrating endometriosis. Molecular Pain. First published date: May-07-2017. 10.1177/1744806917705564.
Figure: Immunohistochemical staining of TRPV1 receptor in healthy eutopic endometrium and in rectosigmoid DIE nodules. (a) Negative control using tris-buffered saline instead of the primary antibody in normal endometrial tissue. (b) Rectal myenteric ganglia, serving as positive control for TRPA1 expression. (c) Healthy eutopic endometrial tissue. (d) Rectosigmoid DIE nodule. (e) Rectosigmoid DIE nodule, glandular component. (f) Rectosigmoid DIE nodule, stromal component. (d) and (f) Sections shown on panels were taken from the same DIE patient who experienced severe, endometriosis associated pain. Background staining was performed with hematoxylin and eosin to reveal the tissue structure. Black arrow heads denote TRPV1 receptor labelling. Magnification is X400, except panel (d) where it is X100. Scale bars: 50 µm, except panel (d) where it is 200 µm.
We frequently post unique data generated by use of our antibodies.

Thursday, May 04, 2017

Culturing Stem Cells in 3-D

Requires Potent Media + Supplements

Neuromics is responding to the many challenges our clients face in building 3-D, in-vivo like, cell- based assays. We do this by offering the most potent Media plus Supplements like FGFS.

Here's a protocol for single cell 3-D assays using hMSCs and Hydrogels. It features use of our ISOKineTM FGF

Images: Cell-centering in cytocompatible microgels enables long-term single-cell 3D culture by preventing cell escape. a) Qualification of Dex-TA microgel crosslinking as a function of the microemulsion flow rate (Qemulsion) and concentration of the H2O2 feed ([H2O2]feed). Blue, green, and red indicate incomplete crosslinking, complete crosslinking, and H2O2 excess, respectively. b,c) Amplex Red assay to quantify the concentration of residual H2O2 ([H2O2]emulsion) in Dex-TA microgel precursor droplets and crosslinked microgels after their retrieval from the diffusion-based crosslinking platform. d) The microencapsulation procedure had no detrimental effect on short-term cell survival. e) Delayed crosslinking resulted in 4 ± 1% cell escape after 7 d of in vitro culture, as compared to 27 ± 5% cell escape when using coupled emulsification and gelation. f) The number of encapsulated cells per microgel tightly followed the Poisson distribution and remained similar throughout long-term (28 d) of in vitro culture, which confirmed that cell centering prevents cell escape. g–i) MSCs encapsulated in delayed enzymatically crosslinked microgels remained viable and metabolically active throughout 28 d of in vitro culture. j) Positive Oil Red O and k) Alizarin Red staining confirmed that l) more than 60% of the microencapsulated MSCs could differentiate into the adipogenic and osteogenic lineage, respectively. Black scale bars: 50 µm, white scale bars: 5 µm. DOI: 10.1002/smll.20160371.

Protocol for Cell Isolation and Expansion: Human MSCs were isolated from fresh bone marrow samples and cultured as previously described. The use of patient material was approved by the local ethical committee of the Medisch Spectrum Twente and informed written consent was obtained for all samples. In short, nucleated cells in the bone marrow aspirates were counted, seeded in tissue culture flasks at a density of 500 000 cells cm−2, and cultured in MSC proliferation medium, consisting of 10% FBS, 100 U mL−1 penicillin, 100 mg mL−1 streptomycin, 1% GlutaMAX, 0.2 × 10−3 m ascorbic acid, and 1 ng mL−1 bFGF (added fresh) in αMEM. Mouse insulinoma MIN6-B1 cells (provided by Dr. P. Halban, University Medical Center, Geneva, Switzerland) were cultured in MIN6 proliferation medium, consisting of 10% (v/v) FBS, 100 U mL−1 penicillin, and 100 mg mL−1 streptomycin, and 71 × 10−6 m 2-mercaptoethanol (added fresh) in DMEM. When cells reached near confluence, the cells were detached using 0.25% Trypsin-EDTA at 37 °C and subsequently subcultured or used for experimentation.

I am at your beck and call to answer questions on our Cell Based Assay Solution. Pete Shuster-CEO and Owner, direct phone: (612) 801-1007 or pshuster@neuromics.com.