Tuesday, April 26, 2016

ELISA Blocking Buffers

Paramount to Reducing Background Noise and Attaining an Accurate Signal

Once the antibody or protein antigens have been properly adsorbed onto the ELISA/EIA plate, the next critical step in creating a reliable immunoassay is the blocking of the plate. This key is:

  • Preventing nonspecific binding 
  • Reducing ELISA background signal 
  • Blocking nonspecific binding to adsorbed proteins 
  • Stabilizing proteins adsorbed to plate for better interactions 



Neuromics's ELISA Blocking Buffer formulations reduce nonspecific binding of sample and assay components to the ELISA well while stabilizing the coated protein. Six formulations offer significant benefits to different assay situations. Browse the product selections below, or order our Blocking Buffer Optimization Pack to try three formulations at an economical rate.

Friday, April 15, 2016

Isolating and Selecting T-Cells

Save 100 USD on New Kits

We are pleased to announce the addition of our New T-Cell Isolation/Selection Kits. They are fast and easy. Check out the Short Protocol.

Figure CD4+ T-Cell Isolation Process doi:10.1371/journal.pone.0035798.

We also have T-Cell Expansion Kits!
NameCatalog #TypeSpeciesApplicationsSizePrice
StrepMan Magnet for 15ml and 50ml Tubes6-5650-065Cell Isolation-SeparationHCell Assays1 Magnet$400
CD3 Fab Streptamer Isolation Kit MB6-8000-201Cell Isolation-SeparationH1 Kit$699
CD4 Fab Streptamer Isolation Kit MB6-8000-206Cell Isolation-SeparationH1 Kit$699
CD8 Fab Streptamer Isolation Kit MB6-8000-203Cell Isolation-SeparationH1 Kit$699
CD25 Fab Streptamer Isolation Kit MB6-8000-207Cell Isolation-SeparationH1 Kit$699
We'll be adding more and more kits in the coming months and will keep you posted.

Saturday, April 02, 2016

Magic Red and Cancer Research

Measure Apoptosis in Real Time in Cancer Cells

Magic Red™ (MR) Kits measure apoptosis via active caspases and cathepsins in whole living, intact cells - no lysis required. As apoptosis progresses and caspase activity increases, the red fluorescent signal increases.

In this example, Magic Red is used to measure apoptosis in HeLa Cells after treatment with Artesunate (ART), an anti-malarial drug and potential chemotherapy agent.



Figure: ART activates lysosomal function. A, HeLa cells were first treated with ART (50 μM) for 12 h, followed by LTR labeling for 30 min, and further analyzed using confocal microscopy. B, HeLa cells were loaded with LTR following time course drug treatment, and fluorescence intensity of 10,000 cells/sample was measured by flow cytometry. C, HeLa cells were treated as indicated in A and then incubated with Magic Red cathepsin L and LTG for 30 min and observed under a confocal microscope. D, HepG2 cells were treated as in A and then labeled with LTR described earlier in A. E, HepG2 cells were treated as in A, followed by a Magic Red cathepsin B and L assay for 45 min. Fluorescence intensity of 10,000 cells/sample was measured using flow cytometry. F, HeLa cells were first loaded with DQ Red BSA for 1 h and then treated with ART (50 μM) for 18 h and incubated with LTG for 30 min. G, HeLa cells were treated with ART for 18 h and then immunostained for LAMP1. H, LAMP1 signals of 80 cells from three independent experiments were quantified by ImageJ (Student's t test, #, no significance). I, HeLa cells were treated with ART for the indicated time, and then Western blot was performed to detect the LAMP1 protein level. Scale bar, 10 μm. Error bars, S.D. doi: 10.1074/jbc.M114.564567

We have a variety of Apoptosis Kits for testing potential Chemotherapeutic Agents.

Thursday, March 31, 2016

Human Astrocytes

Cortex Derived Astrocytes

Neuromics is pleased to be offering yet another option for culturing Human Astrocytes. They can be passaged up to 10X-Only 749 USD/500,000 Cells

They are deigned to be easy to culture and grow.

 Image: Human Brain Astrocytes cultured with our AlphaBioCoat.
Check out our large offering of Neurons, Astroglia, Progenitors, Brain Endothelial Cells/Pericytes and Blood Brain Barrier (BBB) Model. Should you have questions on these or any of our offerings, I can be reached directly at 612-801-1007 or pshuster@neuromics.com. Thank you, Pete Shuster-CEO and Owner.

Thursday, March 24, 2016

Epigenetics and Pain Research

i-Fect Used to Study Impacts

Our i-Fect siRNA, miRNA and shRNA Trasfection Kit was recently used to study the impact of G9a-specific siRNA (AGUAACGGGCAUCAAUGC) on Mu Opioid Receptors: Yuhao Zhang, Shao-Rui Chen, Geoffroy Laumet, Hong Chen and Hui-Lin Pan. Nerve Injury Diminishes Opioid Analgesia through Lysine Methyltransferase-Mediated Transcriptional Repression of µ-Opioid Receptors in Primary Sensory Neurons. First Published on February 25, 2016, doi: 10.1074/jbc.M115.711812... In some SNL rats, G9a-specific siRNA (4 µg) or the negative control siRNA was administered intrathecally. G9a-specific siRNA(AGUAACGGGCAUCAAUGC) or universal negative control siRNA (#SIC001, Sigma-Aldrich) was mixed with i-Fect (Neuromics, Edina, MN) to a final concentration of 400 mg/L for the intrathecal injections...

Figures: G9a knockdown with siRNA reverses the MOR expression in the DRG and the morphine analgesic effect diminished by nerve injury. (A,B) Quantitative PCR (A) and Western blotting (B) analyses show the mRNA and protein levels of MORs in the DRGs of sham and SNL rats treated with control or G9a-specific siRNA (n = 10 rats in each group). The ipsilateral L5 and L6 DRG tissues were removed 24 h after the last siRNA injection. The amount of MOR mRNA and protein was normalized to GAPDH in the same samples, and the mean value of MOR levels in sham control rats was considered to be 1. (C) Time course of the intrathecal morphine effects on the tactile and pressure withdrawal thresholds in sham and SNL rats treated with G9a-specific siRNA or negative control siRNA (n = 9 rats in each group). The withdrawal thresholds after the last siRNA injection were plotted as the baseline control (BL).

Summary: The findings provide new insight into the epigenetic mechanism regulating MOR expression in primary sensory neurons in neuropathic pain. This multidisciplinary approach provides conclusive evidence for G9a as a key chromatin regulator responsible for MOR downregulation in the DRG and the analgesic efficacy of opioids reduced by nerve injury. A better understanding of the epigenetic mechanisms underlying nerve injury-induced downregulation of MORs in primary sensory neurons could help improve the analgesic efficacy of opioids for treating chronic neuropathic pain. G9a inhibitors could be used to enhance the opioid analgesic effect and reduce opioid consumption in patients with chronic neuropathic pain.

Tuesday, March 15, 2016

3-D Tissue Models Made Easy

3 Steps to an in vivo Like Assay
We provide both CollaGel Hydrogels with different stiffness options and ready made 3-D Human Tissue Models.

 Building these models can be done in 3 easy steps:
  • Step 1: Thaw our CollaGel Hydrogel solution 
  • Step 2: Add your cell suspensions 
  • Step 3: Watch it gel
HUVECS intergrated into 3-D CollaGel environment.

Due to the way in which it gels, our Collagel Hydrogel is well suited to Bio Printing models.

I will be posting more data as it becomes available.

Saturday, March 12, 2016

CRISPR Everywhere

Nature Explores What it Means

Here's the link: http://www.nature.com/news/crispr-everywhere-1.19511
We are part of the CRISPR everywhere revolution!

Features Include:

  • Simple-"Cut and Paste" Integration and Editing. 
  • Check out our Sleeping Beauty Transposon System Users Manual 
  • Potent-100% Stable with No Off target effects with the safest transposon insertion profile. 
  • Fast- Transfect with our transposase + Sleeping Beauty TransposonTM Reporter Series Vectors. 
  • Cost Effective- Starting at $329

Friday, March 04, 2016

Check Out Our 3D Human Skin Tissue Model

In vivo Like Model for Studying Skin Diseases/Irritants

I am pleased to announce this new addition to our Cell Based Assay offerings.

3D Human Skin Tissue Model
Cross section of Human Skin Model
The model includes precultured tissue in our engineered and tissue/cell specific Collagel-Hydrogels.

Our 3D human skin model is manufactured in a GMP lab. The model is engineered from multilayered, differentiated epidermal cells. The tissue is provided cultured in inserts using serum free media. The model mimics human skin. It provides an in vivo like environment for studying dermal related diseases and cancers. It is also excellent for skin toxicology and irritancy assays.

Check all our  3D Models:
Name Applications Size Price
Human Blood Brain Barrier Model In vivo Like Assays 6 wells
12 well
24 well
$1,780
$2,600
$3,625
3D HUVEC Model In vivo Like Assays 6 well
12 well
24 Well
$1,269
$1,399
$1,539
3-D Human Skin Tissue Model In vivo Like Assays 6 well
12 well
24 Well
$1,269
$1,399
$1,539

Questions? Contact me directly-Pete Shuster, CEO and Owner, 612-801-1007 or pshuster@neuromics.com.

Monday, February 22, 2016

Stem Cell "Stemness" and Differentiation States

Markers Play a Key Role

Our Stem Cell Markers are widely used and frequently published. These play an important role for understanding the molecular events that take place during the differentiation of human pluripotent cells. They are essential for the protocols aimed at insuring the generation high of quality differentiated cells.

Here's an excellent study on the determining the state of differentiating stem cells: Julio Castaño, Cristina Morera, Borja Sesé, Stephanie Boue, Carles Bonet-Costa, Merce Martí, Alicia Roque, Albert Jordan, Maria J. Barrero. SETD7 Regulates the Differentiation of Human Embryonic Stem Cells. Published: February 18, 2016DOI: 10.1371/journal.pone.0149502...SOX2 (Neuromics GT15098)...

Figure: SETD7 is expressed at very low levels in pluripotent human cells and induced during differentiation. (A) Average rank of the top 700 most differentially expressed genes between pluripotent (iPSCs or ESCs) and fibroblasts, including those upregulated in pluripotent cells (left panel) and upregulated in fibroblasts (right panel). (B) SETD7 mRNA levels in human ESCs grown under self-renewal conditions (UndES[4]), in vitro differentiated human ESCs (DifES[4]), human fibroblasts (HFF), two lines of human keratinocytes (HEK1 and HEK2) and two lines of iPSCs generated from keratinocytes ([H]KiPS4F and KiPS4F1). Mean and standard deviation of three technical replicates is shown. Induction of SETD7 mRNA levels during ES[4] differentiation was confirmed in more than four independent differentiation experiments. (C) Western blot showing SETD7 protein levels in pluripotent and somatic cells. Loading control beta actin (ACTB) is also shown. (D) Western blot showing protein levels of SETD7, AFP, OCT4 and SOX2 in under self-renewing conditions and in vitro differentiated human ESCs. Loading control alpha tubulin (TUBA) is also shown. One representative experiment out of three is shown. (E) Genomic visualization of the levels of H3K72me3, H3K4me3, H3K4me2, H3K36me3 and RNA polymerase II (Pol II) in the human embryonic stem cell line H1 around the SETD7 gene according to ENCODE. A non-methylated CpG island is depicted in green. (F) Levels of H3K4me2 and H3K27me3 at SETD7 gene promoter region (27 bp upstream of the transcription start site) in pluripotent and somatic cells determined by chromatin immunoprecipitation (ChIP) and ploted relative to the input. IgGs wer used as negative control. Bars show the mean and standard deviation of three independent immunoprecipitations...doi:10.1371/journal.pone.0149502.g001

Neuromics is pleased to provide many different stem cell related options for your cell based assays. I will continue to post updates.

Monday, February 15, 2016

3-D Cell Based Assays Are Evolving

3-D Multicell Models and Gels

There is a growing body of publications of why 3-D matters in drug discovery and toxicology assays. This table shows important distinctions for single cell assays.
Cellular characteristics 2D 3D Refs.
Morphology Sheet-like flat and stretched cells in monolayer Natural shape in spheroid/aggregate structures 20,24,50
Proliferation Often proliferate at a faster rate than in vivo May proliferate at a faster/slower rate compared to 2D-cultured cells depending on cell type and/or type of 3D model system 17,51
Exposure to medium/drugs Cells in monolayer are equally exposed to nutrients/growth factors/drugs that are distributed in growth medium Nutrients and growth factors or drugs may not be able to fully penetrate the spheroid, reaching cells near the core 24,52
Stage of cell cycle More cells are likely to be in the same stage of cell cycle due to being equally exposed to medium Spheroids contain proliferating, quiescent, hypoxic and necrotic cells 18,24,53
Gene/protein expression Often display differential gene and protein expression levels compared to in vivo models Cells often exhibit gene/protein expression profiles more similar to those in vivo tissue origins 17,40,54
Drug sensitivity Cells often succumb to treatment and drugs appear to be very effective Cells are often more resistant to treatment compared to those in 2D culture system, often being better predictors of in vivo drug responses 17,33
Table: Key Differences in Cellular Characteristics and Processes in Two-Dimensional and Three-Dimensional Culture Systems

From the table, we see, for the most part, the overall advantages of cells in 3-D for generating more in vivo like data. We also see potential issues like: "Nutrients and growth factors or drugs may not be able to fully penetrate the spheroid, reaching cells near the core".  

We have resolved penetration issues with our engineered Collagel Hydrogels. We have further enhanced the assays by including multicell assays that even more closely mimic the in vivo environment.

Our Blood Brain Barrier (BBB) Model, for example, includes our:
These cells are pre-cultured in the engineered gels and cost a fraction of the individual components (see: culturing and assay protocol).
Name Catalog #

Size Price
Human Blood Brain Barrier Model 3D45002-6 6 wells
12 well
24 well
$1,780
$2,600
$3,625
Our intention is for you to generate the most in vivo like data possible.

Tuesday, February 09, 2016

Ready to Use Blood Brain Barrier Model (BBB)

6, 12 and 24 Well Options

We are pleased to introduce our BBB Model. It is designed to provide more in vivo like data for your drug discovery and toxicology assays.

Our model mimic transport properties of the BBB due to the formation of tight junctions, higher expression of specific carriers, or great cell viability. The model includes brain endothelial cells with pericytes and astrocytes layered in an insert. This model improves endothelial cell polarization and enhance the formation of tight junctions, provide better endothelial cell-to-cell contact that is important for barrier development, and prevent the dilution of secreted neurotrophic factors, and these conditions collectively led to the development of an in vitro model that can truly mimic the BBB.
Diagram of BBB Model Transporters
Transport of a molecule (blue) through 3D Human BBB Model in a luminal to basal direction 
Calculation of apparent permeability (Papp in cm/min)
Apparent permeability coefficient Papp (in cm/min) based on the Fick's law can be calculated according to the following equation:
VA: volume of abluminal chamber (cm3)
A: membrane surface area (1.12 cm2)
[C]L: initial luminal tracer concentration (ng/ml)

[C]A : abluminal tracer concentration (ng/ml) t : time of experiment (min)

 Our goal is to provide you better data...at a lower cost.

Sunday, January 24, 2016

More and More OR Pubs

Potent, Proven and Frequently Published

Many labs have used our Opioid Receptor antibodies over the past 10+ years. We are grateful for the many publications and testimonals referencing successful use of these antibodies.

Here's a sampling of more recent publications: Wendy M. Walwyn, Wenling Chen, Hyeyoung Kim, Ani Minasyan, Helena S. Ennes, James A. McRoberts, and Juan Carlos G. Marvizón. Sustained Suppression of Hyperalgesia during Latent Sensitization by μ-, δ-, and κ-opioid receptors and α2A Adrenergic Receptors: Role of Constitutive Activity. The Journal of Neuroscience, 6 January 2016, 36(1): 204-221; doi: 10.1523/JNEUROSCI.1751-15.2016
...probed with antibodies to MOR (RA10104; Neuromics), p-Ser375-MOR (pMOR, RA18001; Neuromics) and p-Tyr416-Src family kinase (pSFK...serine-phosphorylated MOR (p-Ser375-MOR, RA18001; Neuromics) and a different MOR antibody (RA10104...
Jan Fraessdorf, Markus W. Hollmann , Iris Hanschmann, André Heinen, Nina C. Weber, Benedikt Preckel, Ragnar Huhn. Role of Endogenous Opioid System in Ischemic-Induced Late Preconditioning. Published: July 30, 2015DOI: 10.1371/journal.pone.0134283.
...Antibodies for μ-OR and δ-OR were purchased from Neuromics (Minneapolis, USA)...
Ying-Biao Chen, Fen-Sheng Huang, Ban Fen, Jun-Bin Yin, Wei Wang and Yun-Qing L nhibitory effects of endomorphin-2 on excitatory synaptic transmission and the neuronal excitability of sacral parasympathetic preganglionic neurons in young rat. Journal Name: Frontiers in Cellular Neuroscience, ISSN: 1662-5102, Article type: Original Research Article,Received on: 02 Apr 2015, Accepted on: 12 May 2015, Provisional PDF published on: 12 May 2015.
... guinea pig antiserum against MOR (1:1000, GP10106, Neuromics, Edina, Minnesota, USA)...
Sherika N. Smith, Candler Paige, Kandy T. Velazquez, Terika P. Smith, Srinivasa N. Raja, Steven P. Wilson, Sarah M. Sweitzer. Injury-specific promoters enhance herpes simplex virus mediated gene therapy for treating neuropathic pain in rodents. doi:10.1016/j.jpain.2014.12.007
...: MOR (Neuromics, RA10104, 1:1000)...


Figure: MOR in primary sensory neuron and its terminals were decreased in neuropathic mice. (A, B) Western blot shows that MOR protein were markedly decreased in sciatic nerve (A) and hindpaw skin (B) on day 7 after nerve injury when compared with sham-operated mice. The ratio of MOR/GAPDH in sham mice was set at 1 for quantifications. Zhou et al. Molecular Pain 2014, 10:51 http://www.molecularpain.com/content/10/1/51

We will continue to post information on our ORs.


Thursday, January 14, 2016

Why You Should Consider Culturing Your Cells in 3-D

2-D versus 3-D and meaningful outcomes

We have seen recent growth in our customers' demand for 3-D cell based assay solutions. This could be due to more the fact that these assays our proving to generate more in vivo like data.

Breast cancer cells, for example, grown in 2-D can easily be killed by low doses of chemotherapeutic drugs or low doses of radiation. If those same cells are grown in 3-D, they are resistant to the same doses of chemotherapeutic drugs or radiation, just like cancer as its found in the body. In this way, cells grown in 3D are more valid targets for testing and discovering new drugs to treat cancer. Another benefit of testing drugs in three dimensional cell culture versus two dimensional cell culture is that cells in 3D form multi-layers of cells whereas cells grown in 2-D form a monolayer of cells that is spread very thin on a plastic surface. When testing a drug in 2-D, it needs only to diffuse a short distance across the cell membrane to reach its intended target. In 3-D, the situation is more realistic and a drug needs to diffuse across multi-layers of cells to reach the cells on the inside of a microtissue.

Neuromics has a variety of options that will enable you to choose the best option(s) for your 3-D assays. These include:
Look for more posting on these important solutions.

Saturday, January 09, 2016

Sleeping Beauty (SB) Transposon for the Study of Cancers

SB Gene Insertion vs Viral Based Methods

With the addition of SB Transposon Systems to our tool set. We have deep interest in how well SB works vs alternative methods. Here Dr. Hyun-Pyo Kim and team demonstrate how the SB Transposon system can be used for Novel Therapeutic Approaches for Various Cancer Types Using a Modified Sleeping Beauty-Based Gene Delivery System (DOI: 10.1371/journal.pone.0086324).

SB vs Viral Based Methods: "Successful gene therapy largely depends on the selective introduction of therapeutic genes into the appropriate target cancer cells. One of the most effective and promising approaches for targeting tumor tissue during gene delivery is the use of viral vectors, which allow for high efficiency gene delivery. However, the use of viral vectors is not without risks and safety concerns, such as toxicities, a host immune response towards the viral antigens or potential viral recombination into the host's chromosome; these risks limit the clinical application of viral vectors. The Sleeping Beauty (SB) transposon-based system is an attractive, non-viral alternative to viral delivery systems. SB may be less immunogenic than the viral vector system due to its lack of viral sequences. The SB-based gene delivery system can stably integrate into the host cell genome to produce the therapeutic gene product over the lifetime of a cell. However, when compared to viral vectors, the non-viral SB-based gene delivery system still has limited therapeutic efficacy due to the lack of long-lasting gene expression potential and tumor cell specific gene transfer ability. These limitations could be overcome by modifying the SB system through the introduction of the hTERT promoter and the SV40 enhancer. In this study, a modified SB delivery system, under control of the hTERT promoter in conjunction with the SV40 enhancer, was able to successfully transfer the suicide gene (HSV-TK) into multiple types of cancer cells. The modified SB transfected cancer cells exhibited a significantly increased cancer cell specific death rate. These data suggest that our modified SB-based gene delivery system can be used as a safe and efficient tool for cancer cell specific therapeutic gene transfer and stable long-term expression."


Figure 8. The effect of modified SB system on the tumor growth in vivo. Lung cancer cells (H358) (A), prostate cancer cell line (DU-145) (B), and ovarian cancer cells (OVCAR3) (C) were harvested by trypsinization, and 1×105 viable cells (as determined by trypan blue exclusion) in a total volume of 200 µl were injected subcutaneously. Two days following tumor seeding, animals were intravenously injected via tail veins with 100 mg/kg gancyclovir (GCV) along with either co-transfection of the empty plasmid (pT. hTp. Con) with the active helper plasmid (pCMV-SB) or co-transfection of the SB system (pT.hTp.HSV-tk.Con) with the active helper plasmid (pCMV-SB). Mice were sacrificed 28 days after tumor injection, and the effect of modified SB system on tumor growth was evaluated by measuring tumor size. doi:10.1371/journal.pone.0086324.g008

Neuromics, with our partner B-Mogen, has the capabilities to engineer custom SB Transposons for your Cancer Research. If interested, please contact me directly at 612-801-1007 or pshuster@neuromics.com. Our process is to first completely understand your unique requirements and from these, formulate a related statement of work with costs, timeline, milestones and deliverables. Thank you. Pete Shuster, CEO and Owner,

Sunday, January 03, 2016

CRISPR-Cas9 and Sleeping Beauty Transposons

Neuromics-B-Mogen Approach

The CRISPR-Cas9 gene editing approach is now officially considered a revolution. This is confirmed by the increasing frequency of this word being used across the web in describing this technique. In fact, it is now appearing in conference titles. see: Genome Engineering: The CRISPR/Cas Revolution 2016COLD SPRING HARBOR - 17 AUG 2016.

We have joined the revolution with our new Sleeping Beauty TransposonTM Systems.
Figure: Sleeping Beauty TransposonTM Systems Gene Integration Process. (A) A depiction of Sleeping Beauty transposon plasmid and Sleeping Beauty transposase enzyme active in cellular nuclei. (B) Transposase enzyme binds to Sleeping Beauty-specific IR/DR sites. (C) Transposase enzyme excises transposon sequence from transposon vector. (D) Transposase enzyme stably integrates transposon sequence at TA site in host cell genome.

Features Include:
  • Simple-"Cut and Paste" Integration and Editing. Check out our Sleeping Beauty Transposon System Users Manual 
  • Potent-100% Stable with No Off target effects with the safest transposon insertion profile. 
  • Fast- Transfect with our transposase + Sleeping Beauty Transposon Reporter Series Vectors. 
  • Cost Effective- Starting at $329.
If you would like us to engineer custom Transposons or gene editing on your cells, please conatact me directly and we will work together to formulate a Statement of Work to your specifications. Pete Shuster, CEO and Owner, Neuromics, pshuster@neuromics.com or cell phone: 612-801-1007

Thursday, December 17, 2015

Fibroblasts into Neurons

Via Activation of Oct4

We have a variety of stem cell, progenitor and neuron markers that are often referenced in publication. There represent a qualitative way to determine the state of stem cells as they differentiate.

Here're researchers take Fibroblasts and differentiate them into various progenitors. They were able to further differentiate these progenitors into astrocytes and neurons by: For neuronal differentiation, after 2 days of incubation with 20 nM reversine, cells were then treated with 0.5 μM all-trans-retinoic acid (RA) in serum-free DMEM/F-12 medium supplemented with the ITS for 2 days and switched into serum-free medium in the absence of RA with replacement of the medium every 2-3 days. 

Our NSE and GFAP were used to confirm this differentiation.
To learn more see: Xiangchen Li, Yu Guo, Yaxin Yao, Jinlian Hua, Yuehui Ma, Changqing Liu, Weijun Guan.Reversine Increases the Plasticity of Long-Term Cryopreserved Fibroblasts to Multipotent Progenitor Cells through Activation of Oct4. International Journal of Biological Sciences 2016; 12(1): 53-62. doi: 10.7150/ijbs.12199

Wednesday, December 09, 2015

Kv Channels and Pain Transmissiom

i-Fect TMis a Proven Tool for Gene Manipulation in Studying All Types of Pain

I previously posted on use of our i-Fect Transfection Kit to silence Kv Channels Receptors. This has enabled researchers to study the role of these receptors in vitro and in vivo (see:i-Fect™ Delivers Your siRNA Payload).

Sample Data

Figure: Figures. siRNA-mediated knockdown of Kv1.1 expression in thoracic DRG significantly increased gastric sensitivity in naive adult rats. (A) Western blots showed a significant decrease in Kv1.1 protein in thoracic DRG (T8–T12) after intrathecal treatment with Kv1.1 siRNA but not with control siRNA. siRNA treatment did not alter TrpV1 expression (n = 5 rats each; *P < .01 vs control siRNA). (B) Naive rats treated with Kv1.1 siRNA showed a significant increase in VMR to gastric distention (n = 5 rats each, compared with pretreatment baseline; *P < .05). (C) Treatment with control siRNA had no significant effect on gastric hypersensitivity. (D) Patch clamp recordings from freshly dissociated gastric DRG neurons from FD-like and PND 10 saline-treated littermate controls showed a significant decrease in rheobase in FD-like rats (*P < .05), and (E) a significant increase in the number of action potentials elicited by current injection at 3× the rheobase in gastric DRG neurons from FD-like rats (*P < .05). (F) Sample voltage vs time traces showing action potentials evoked at ×1, ×2, and ×3 rheobase. The patch clamp data were obtained from 16 cells from 5 PND 10 saline control rats and 19 cells from 5 FD-like rats

I am pleased to share with you a new reference detailing how research use i-Fect to optimize and deliver euchromatic histone-lysine N-methyltransferase-2 (G9a) siRNA. This brings the number of publications referencing use of our Transfection Kits to over 45: Geoffroy Laumet, Judit Garriga, Shao-Rui Chen, Yuhao Zhang, De-Pei Li, Trevor M Smith, Yingchun Dong, Jaroslav Jelinek, Matteo Cesaroni, Jean-Pierre Issa & Hui-Lin Pan G9a is essential for epigenetic silencing of K+channel genes in acute-to-chronic pain transition. Nature Neuroscience (2015) doi:10.1038/nn.4165.

The authors report: "Selective knockout of the gene encoding G9a in DRG neurons completely blocked K+ channel silencing and chronic pain development after nerve injury. Remarkably, RNA sequencing analysis revealed that G9a inhibition not only reactivated 40 of 42 silenced genes associated with K+ channels but also normalized 638 genes down- or upregulated by nerve injury."

I will continue to post here new and unique solutions and related referencing for our Gene Expression Analysis Tools.

Saturday, November 28, 2015

Neuron Astrocyte Glial Markers

Potent and Frequently Published!

We have a stout catalog of Neuron-Astrocyte and Glial Markers.

They are widely used and frequently published. Here're some recent examples:
Mouse Monoclonal GFAP: Chelsea M. Larabee, Constantin Georgescu, Jonathan D. Wren and Scott M. Plafke. Expression profiling of the ubiquitin conjugating enzyme UbcM2 in murine brain reveals modest age-dependent decreases in specific neurons. BMC Neuroscience201516:76 DOI: 10.1186/s12868-015-0194-y© Larabee et al. 2015.

Image: Mixed neuron-glial cultures stained with Mouse Monoclonal GFAP, and Chicken Polyclonal Neurofilament-NF-L (green). The GFAP antibody stains the network of astrocytes in these cultures, while the NF-L antibody stains neurons and their processes. The blue channel shows the localization of DNA. This antibody also works on formalin fixed paraffin embedded brain tissues. Protocol on Datasheet.

Tuj-1: Gaoying Sun, Wenwen Liu, Zhaomin Fan, Daogong Zhang, Yuechen Han, Lei Xu1, Jieyu Qi, Shasha Zhang, Bradley T. Gao, Xiaohui Bai,Jianfeng Li,Renjie Chai, Haibo Wang. The three-dimensional culture system with matrigel and neurotrophic factors preserves the structure and function of spiral ganglion neuron in vitro.

Need more assurances? Here're some feedback highlights.

STEPHEN C. Nov 19, 2015 Staining (Tuj1) worked well on human neural progenitor cells. Product Name: Tuj 1, Mouse – (Cat# MO15013-100) http://bit.ly/1Qx4ASc Organization: UCONN
GINA D. Oct 07, 2015 Very nice antibody and ordering is very easy using the website. Product Name: proDynorphin (rat), Guinea Pig – (Cat# GP10110) http://bit.ly/Sw4RJ9 Organization: Rosalind Franklin University
CAROL Feb 05, 2015 We got antibodies and received them fast with a correct temperature. Product Name: Coronin 1A, Chicken – (Cat# CH23017) http://bit.ly/1AxduYvProduct Name: Integrin alpha-M, Chicken – (Cat# CH23021)http://bit.ly/16lMihU Organization: UCSF

We have a full money back guarantee so do not hesitate to consider these markers for your assays.

Friday, November 20, 2015

Staining Neurons and 3-D

Tuj-1 or Beta Tubulin Antibody in Action!

Our potent Neuron-Glia Markers are widely used and frequently published. We are also pleased with the many positive reviews.

In this study researchers use our Tuj-1/Beta Tubulin Antibody to stain neurons in 3-D Cultures: Gaoying Sun, Wenwen Liu, Zhaomin Fan, Daogong Zhang, Yuechen Han, Lei Xu1, Jieyu Qi, Shasha Zhang, Bradley T. Gao, Xiaohui Bai,Jianfeng Li,Renjie Chai, Haibo Wang. The three-dimensional culture system with matrigel and neurotrophic factors preserves the structure and function of spiral ganglion neuron in vitro...Tuj-1-β-tubulin (1:1000, Neuromics, USA)...

Images: Morphology of the SGNs growth cone cultured in 2D and 3D systems. Phalloidin, green; β-tubulin, red.

I will continue to post positive developments concerning use of our Neuron-Glia Markers.

Wednesday, November 11, 2015

Reconstructing 3D Living Biomaterials

Hydrogel Solutions for Tissue Reconstruction and Repair

Alphabioregen/Neuromics are pleased to announce the use of our Hydrogels for reconstructing living biomaterials for regenerative biology and medicine: Suwan N. Jayasinghe,Jensen Auguste, Chris J. Scotton. Platform Technologies for Directly Reconstructing 3D Living Biomaterials. DOI: 10.1002/adma.201503001.

In this study, researchers use our  Collagen Hydrogel, Collagen Hydrogel Soft and Collagen Hydrogel Soft+  to describe  in vivo applications using a murine model to interrogate biocompatibility and cellular behavior post-transfer.

Luc macrophages (CC or BES) were then incorporated into a fl uid mixture containing one of three alternative collagen-based hydrogels, namely, Collagen Hydrogel, Collagen Hydrogel Soft, or Collagen Hydrogel Soft+ (resulting in six possible combinations of macrophages and biopolymer).

Overview of Results



Figure: Bioluminescent imaging of implanted macrophage–biopolymer mixes. IC-21-Luc macrophages (CC or BES) were mixed with biopolymer (Hydrogel, Soft, or Soft+) and subcutaneously injected into the dorsal fl anks of C57Bl/6 mice (three mice per hydrogel, with CC on the left fl ank and BES on the right). Following intraperitoneal injection of delta-luciferin, macrophage bioluminescence was detected using an IVIS Lumina II imaging system. A representative image from day 1 post-implantation (and 25 min post luciferin injection) is shown, indicating the peak detectable radiance (photons s −1 cm −2) in identically sized regions of interest. The CE results were very similar to the BES implants.


Figure:  Histological analysis of the macrophage–biopolymer implantation site (MSB staining). At day 4 post-implantation, skin was harvested from each dorsal fl ank, and processed for histology. Where possible, serial sections directly adjacent to those shown in Figure 4 were stained with a modifi ed trichrome stain. Mature fi brillar collagen within the dermis appeared dark blue, while the collagen within the hydrogel was generally lighter blue in appearance, indicating a less highly cross-linked or fi brillar form of collagen. Scale bar: 200 µm.

These Collagen Hydrogels are proving easy to use and effective in the hands of our customers. If you are looking for solutions for tissue reconstruction/repair or 3-D in vivo like cell based assays, do not hesitate to contact me @ direct phone: 612-801-1007 or pshuster@neuromics.com. Thank you, Pete Shuster, CEO and Owner, Neuromics.

Saturday, November 07, 2015

Human alpha-Sensory Neurons

Put Them to Work!

We continue to add unique primary Human Neurons to our offerings.

We have discounted our new alpha-Sensory Neurons to make it easier to buy and try-only 850 USD/1,000,000 cells.


Just thaw and culture. Simply culture on ECM- Laminin or Fibronectin coatings with our alpha Motor Neuron Maintaining Media. Questions? Please contact me directly 612-801-1007 or pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics.

Thursday, October 29, 2015

Solutions for Blood Brain Barrier Research

Neuromics Has the Key Components!

We are pleased to add Human Brain Microvascular Pericytes (HBMVPCs) to our solution set for Blood Brain Barrier (BBB) Researchers. These join our Human Brain Microvascular Endothelial Cells and hAstroPro Human Astrocyte+Astroglial-Neurons Co-Culturing Kits to round out our offerings of cells involved in BBB regulation.
 
Pericytes function at the BBB in at least two ways: by regulating BBB-specific gene expression patterns in endothelial cells, and by inducing polarization of astrocyte end-feet surrounding CNS blood vessels (see: http://www.nature.com/nature/journal/v468/n7323/full/nature09522.html).

Questions? Do not hesitate to call me or e-mail me-612-801-1007 or pshuster@neuromics.com). Pete Shuster CEO and Owner.

Tuesday, October 20, 2015

Brain Freeze!

Our Friend are Taking Neuromics Brain on Interesting #brainadventure

It is getting harder to select the winners. You can submit your entry now and earn a chance to win our monthly $100 monthly and $500 grand prize. To enter see: http://www.neuromics.com/brain-adventure-contest.

We can all relate to this.

Tuesday, October 13, 2015

hNP1 Neural Progenitors and Neuronal Regeneration after Injury

The Potential of Axonal PPARγ

In this study, researchers used our hNP1TM Neural Progenitors to demonstrate that axonal PPARγ is involved in neuronal injury responses required for axonal regeneration: Juan Pablo Lezana, Shachar Y. Dagan, Ari Robinson, Ron S. Goldstein, Mike Fainzilber, Francisca C. Bronfman, and Miguel Bronfman. Axonal pparγ promotes neuronal regeneration after injury. DOI: 10.1002/dneu.22353... Culture and regeneration analysis of human axons. NP1 human neural precursors were purchased from Neuromics (Minnesota, USA) and passaged up to 8 times before generation of neurons. Cells were used up until passage 10 after defrosting the purchased neural...

It is important to note that the researchers built up a large stock by passaging the progenitors prior to differentiation.

Check out our hNP1 publications to learn of more unique applications!

Thursday, October 01, 2015

#brainadventure Update

2 Win $100 Monthly Prices!

Our customers are creative. They find interesting #brainadventures for our Neuromics' Brains. Competitive hing for winning our $100 monthly and $500 grand prizes: more entries raise your odds of winning.

Here're some of the submissions:

Learn how you can win @ http://www.neuromics.com/brain-adventure-contest.

Good luick to all.

Saturday, September 26, 2015

Retinal Microvascular Endothelial Cells & Diabetic Retinopathy

HRMECS in Action

Our Human Retinal Microvascular Endothelial Cells in this publication on Diabetic Retinopathy. In this study, the authors show that human retinal microvascular endothelial cells exposed to high levels of glucose regulate miR-200b repression through histone methylation and that inhibition of PRC2 increases miR-200b while reducing VEGF: Michael Anthony Ruiz, Biao Feng, and Subrata Chakrabarti. Polycomb Repressive Complex 2 Regulates MiR-200b in Retinal Endothelial Cells: Potential Relevance in Diabetic Retinopathy. PLoS One. 2015; 10(4): e0123987. Published online 2015 Apr 17. doi: 10.1371/journal.pone.0123987...To investigate PRC2 and miR-200b regulation in the context of diabetic retinopathy, the major cell type used for investigation was the human retinal microvascular endothelial cell (HRMECs,Neuromics/ Olaf Pharmaceuticals, Worchester, MA, Cat# HEC09)...

Figure: High levels of glucose alter VEGF and miR-200b expression in HRMECs. A: HRMECs exposed to various concentrations of D-glucose for 24 hours exhibited differential mRNA levels of VEGF. Compared to 5mM D-glucose, VEGF expression was significantly increased at 15mM and 25mM D-glucose concentrations, with no change at 20mM L-glucose. B: Measured by WST-1 assay, HRMECs exposed to increasing concentrations of D-glucose for 24 hours exhibited decreased cell viability at 25mM, 50mM and 100mM compared to 5mM. C: HRMECs exposed to 25mM (high glucose; HG) glucose for 24 and 48 hours demonstrated significantly increased VEGF mRNA compared to 5mM (normal glucose; NG). These differences were not observed at time points earlier than 24 hours. D,E: HRMECs exposed to 5mM D-glucose (NG) 25mM D-glucose (HG) and 20mM L-glucose+5mM D-glucose (osmotic control; OSM). HRMECs cultured for 24 hours and 48 hours in HG showed significantly decreased levels of miR-200b with parallel increased levels of VEGF expression compared to NG and OSM. F,G: VEGF is also increased at the protein level in HG compared to NG as measured by Western Blotting. [* p < 0.05 compared to NG; n = 6; data expressed as mean ± SEM, normalized to β-actin or U6 and expressed as a fold change of NG]. doi:10.1371/journal.pone.0123987.

Check out our growing catalog of Human Endothelial Cells-Artery, Microvascular, Vein, Umbilical Cord Derived + Culture Media.