Wednesday, May 18, 2016

IGF and Mood

Gender Specific?

IGF is a known mood modulator. This paper hypothesizes that the impact of increased IGF is sex dependent: Victor Munive, Andrea Santi and Ignacio Torres-Aleman. A Concerted Action Of Estradiol And Insulin Like Growth Factor I Underlies Sex Differences In Mood Regulation By Exercise. Scientific Reports 6, Article number: 25969 (2016) doi:10.1038/srep25969.

Our IGF-I R/CD221-Fluorescein Labeled was used to link Estrodial with elevating IGF-I levels in female mice.
Images: (A) Cultured brain endothelial cells accumulate significantly more IGF-I after treatment with 10−10M estradiol (E2). Representative blot is shown (*p less than 0.05 vs control; n = 8 per group). (B) The effects of E2 were mediated by α E2 receptors as only the α inhibitor MPP blocked E2 actions. Representative blots are shown (*p less than 0.05; n = 8). (C) Levels of IGF-I receptor (green) at the cell membrane of endothelial cells are markedly increased three hours after addition of 10−10M E2.

Summary: Sex-specific responses to physiological neuroprotective stimuli such as physical activity, that modulate mood in part through modulation of endocrine signals, contribute to sex differences in mood homeostasis. Hopefully, a better understanding of these differences will help us gain insight of sex differences in the incidence of mood disorders.

Friday, May 13, 2016

Cancer-Associated Fibroblasts and Cancer Cell Migration

Highway for Cancer Metastasis

Cancer Associated Fibroblasts provide the Matrices  (ECMs) serve as cancer’s assistants as it spreads throughout the body.



Cancer cells (red) migrate on a CAF-derived extracellular matrix (green). VANDERBILT UNIVERSITY; BEGUM ERDOGAN, DONNA WEBB

Unlike other ECMs, which normally form dense meshes, these arrange into parallel bundles forming the highway for tumor cell migration (The Scientist-December 15, 2015).

Monday, May 02, 2016

New Neuronal Astroglial PNS Markers

Save 70 USD on High Titer Antibodies
The foundation of our company is built on our catalog of proven, published and high titer Neuron-Astroglia and PNS Markers. We are pleased to offer 75 USD off our latest additions.
Antibody
Type
Species Reactivity
Applications
Mouse IgG
Chicken IgY
Chicken IgY
Mouse IgG
Mouse IgG
Mouse IgG
Mouse IgG
Mouse IgG
Chicken IgY
Rabbit IgG
Mouse IgG
B; H; M; R
B; H; M; R
H; M; R
B; H; M; R
B; H; M; R
H; R
Ch; H; M; Pr; R
Ch; H; M; R
H; M; R
H; M; R
B; H; M; R
IF; WB
ICC; IF; IHC; WB
ICC; IF; IHC; WB
IF; WB
IF; WB
IF; WB
ICC;
IHC; WB
ICC; IHC; WB
ICC; IF; IHC; WB
ICC; IF; IHC; WB

Images: Left: View of mixed neuron/glial cultures stained with Aldolase-C (green) and our rabbit antibody to NeuN/FOX3 (red). MCA-4A9 antibody reveals strong cytoplasmic staining in astrocytes, while Rabbit Fox3/NeuN antibody shows nuclear and distal cytoplasmic staining in neuron cells and is complete absence of astrocytes. Blue is a DNA stain. Middle and Right: Mouse brain sections (fixed by transcardial perfusion with 4% paraformaldehyde) stained with Aldolase-C (red) and our chicken Vimentin antibody (green). In the striatum (Middle), Aldolase-C positive astrocytes are highly co-stained Vimentin, which results in yellow to gold colors. In the cerebellum (Right), however, Aldolase-C positive Purkinje cells do not express vimentin, which results in red color. Insets show a higher magnification picture of MCA-4A9 single labeling in red. Nuclei are labeled with DAPI (blue).
Our clients often use these antibodies for double and triple labeling like the above example.


We plan on aggressively be adding more and more of these type of antibodies. Stay tuned.

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.