Quality, High Titer Neuronal Markers

You Need Them; We got Them

We are recognized for both the number and quality of Neuronal Markers. Check out our many testimonials from satisfied customers.

Here's a sampling of recent publications.

Elisabet Garcia-Pino, Nikodemus Gessele and Ursula Koch. Enhanced Excitatory Connectivity and Disturbed Sound Processing in the Auditory Brainstem of Fragile X Mice. Journal of Neuroscience 3 July 2017, 2310-16; doi.org/10.1523/JNEUROSCI.2310-16.2017. ...chicken α-MAP2 (Neuromics; dilution 1:1000)...
Anna Lisa Gündner Claas Aiko Meyera, Stefan Aigner, Klaus Christensen, Christoph Patsch, Ravi Jagasia, Karlheinz Baumann, Marc Burcin. Generation of a homozygous GBA deletion human embryonic stem cell line. Stem Cell Research. Available online 11 July 2017. https://doi.org/10.1016/j.scr.2017.07.009 ...Neuronal differentiation marker, Ch-MAP2, 1:1000, Neuromics # CH22103 RRID:AB_2314763 ....Immunocytochemistry. Cells were cultured on PO/LAM coated 96-well imaging microplates (Falcon) and fixed with 4% paraformaldehyde (15 min, room temperature). Fixed cells were permeabilized with 0.2% Triton (Sigma) in phosphate-buffered saline (PBS) for 30 min and blocked for 1 h with PBS + 5% donkey serum (Merck Millipore). Primary antibodies (in PBS) were incubated overnight at 4 °C followed by three PBS washing steps. Secondary antibodies were incubated for 2 h at room temperature. Confocal images were acquired using a Leica TCS SP5 microscope (Leica Microsystems)....
Hayk Harutyunyan, Svetlana Sharoyan, Alvard Antonyan, Sona Mardanyan. Herb Preparations Improve the Viability of Hippocampal Cells Suppressed by Amyloid Beta (1-42) Peptide. World Journal of Pharmaceutical Sciences. 2017. ISSN (Online): 2321-3086......Antibodies for immunocytochemistry were purchased from Neuromics (USA)[....]Immunocytochemistry: Hippocampal cells were cultured in Poly-D-Lysine coated Nunc EasY Flasks. On the 4th day, the adhered cells were removed by trypsinisation (0.05% trypsin, 0.5 mM EDTA, pH 8.0). Cells were fixed on microscope slide by 3.7 % paraformaldehyde in PBS followed by methanol treatment. To block a nonspecific antibody binding, samples were pre-treated with goat serum (Sigma) for 30 minutes. To determine the types of cells, constituting the obtained cell culture, the different aliquots of the culture were incubated for 60 min at room temperature with the primary antibodies against rabbit anti-rat Neurofilament (NF), chicken anti-rat GFAP and Nestin...

Images: Cells grown from adult rat brainLarge cell in middle is stained with mouse monoclonal to NF-L clone DA2 (green). Another type of neuronal lineage cell was stained with rabbit polyclonal to alpha-internexin (red). These cells were mitotic but had several characteristics of neurons. Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including: NF-H, NF-M,, NF-L,, NF66 and GFAP 

We will continue to post the latest and greatest news.

Curcumin and Aging Health

Cellular Uptake Matters-Not All Curcumin Based Supplements Are Created Equal

There have been many academic publications on the miracles of Curcumin for Aging Health. Here's a description of its therapeutic properties: The desirable preventive or putative therapeutic properties of curcumin have also been considered to be associated with its antioxidant and anti-inflammatory properties. Because free-radical-mediated peroxidation of membrane lipids and oxidative damage of DNA and proteins are believed to be associated with a variety of chronic pathological complications such as cancer, atherosclerosis, and neurodegenerative diseases, curcumin is thought to play a vital role against these pathological conditions. The anti-inflammatory effect of curcumin is most likely mediated through its ability to inhibit cyclooxygenase-2 (COX-2), lipoxygenase (LOX), and inducible nitric oxide synthase (iNOS). COX-2, LOX, and iNOS are important enzymes that mediate inflammatory processes. Improper upregulation of COX-2 and/or iNOS has been associated with the pathophysiology of certain types of human cancer as well as inflammatory disorders. Because inflammation is closely linked to tumor promotion, curcumin with its potent anti-inflammatory property is anticipated to exert chemopreventive effects on carcinogenesis. Hence, the past few decades have witnessed intense research devoted to the antioxidant and anti-inflammatory properties of curcumin. In this review, we describe both antioxidant and anti-inflammatory properties of curcumin, the mode of action of curcumin, and its therapeutic usage against different pathological conditions see: (Adv Exp Med Biol. 2007;595:105-25).
Read More,,,

Stem Cell Based Therapy For Tendinosis

Using Collagen I Producing Cells from Hair Follicles

Learn how these cells can be these cells can be harvested, purified and expanded to treat a chronic, degeneration of tendons.

Immune System/Stem Cell Health and Aging

Looking, feeling and  performing your best depends on balanced immunity and stem cell vitality

As we age we encounter:

• Slower healing
• Longer recovery time from vigorous exercise
• More general aches and pains
• Longer recovery from illness
• Wrinkled, dry and thinning skin

Why is that? What are the causes? Answers can be found by a better understanding of immune/inflammatory response and stem cell systems. As we age our immune system weakens or becomes dysregulated (autoimmunity) and our stem cell "bank balances" deplete. My friend and world class stem cell therapies expert, Dr. Neil Riordan, gives an excellent description of the process: Your Body’s Stem Cell Bank Account.

To oversimplify, the immune system is responsible for cleansing the body of pathogens, toxins, allergens and damaged tissues/cells. This creates a healthy environment for stem cells to repair and regenerate new, healthy cells and tissue. We call this process the immunoLink^TM.

immunoLink Therapies is a new company of mine slated to launch in the spring of 2015. Our goal is to slow the aging process by offering solutions that balance your immunity and increase your stem cell vitality. Our vanguard product (currently available) is Stem-Kine. It is a blood stem cell booster which increases the level of immune factors and red blood cells in your cardiovascular system. The result is faster healing/recovery and increased endurance via better oxygen delivery.

If you desire to learn more, do not hesitate to call (612-801-1007) or e-mail: pshuster@neuromics.com. Thank you. Pete Shuster

The Dance Between The Immune System and Stem Cells

We named it the  immunoLink^TM 

We have been testing a growing number of Clients with our Quantibody Arrays. Many of of these clients have Autoimmune Disorder Diseases. These range from Rheumatoid Arthritis to Multiple Sclerosis.

These arrays are designed to precisely measure factors or markers (proteins) that are dysregulated by these diseases. We measure the levels of these biomarkers in our Clients' Blood serum. The arrays have also been used to measure the levels of markers in plasma and cell culture supernatants.

Based on results, we are finding links between immune system and stem cell health. We call this the immunoLink. The link shows that when immune/inflammatory response markers are elevated, markers related to stem cell health are depleted.

Here we see the immune/inflammatory response markers IL-6, MCP-1 and TNF-alpha are elevated in our Clients with autoimmunity (A) vs Healthy Controls (HC). We also see lower levels of G-CSF and GM-CSF in these Clients.

G-CSF and GM-CSF are know to play a role in increasing circulating stem cells. GM-CSF is also know to be secreted by Mesenchymal Stem Cells (hMSCs) AND GM-CSF has anti-apoptotic functions on neurons, and is neuroprotective in animal stroke models while G-CSF has a prominent effect on the differentiation of adult neural stem cells (see: BMC Neuroscience 2007, doi:10.1186/1471-2202-8-88).

To us, the immunoLink means achieving a balance between immune system and stem cell health.

We provide immune system balancing and stem cell activating therapies for our Autoimmune Disease Clients and Children with Autism. We first do baseline and follow on testing (each 6 months) to determine how well the therapies are working. Our goal is to bring the many markers we test to healthier levels. As stem cell transplants become more common, moderating levels of immune/inflammatory response in patients could improve outcomes. If you would like to learn more, you can contact me at pshuster@neuromics.com or 612-801-1007.

hNP1 Neural Progenitors Thawing+ Plating Protocol

This is a great video that shows thawing + plating protocol for our eSC Derived Human NP1 Neural Progenitors.

SOX 17 Antibody that Rocks

Our SOX Antibodies are proving to be excellent Stem Cell Markers.  I would like to highlight a pub that references use of our Sox17  and a variety of other markers:

Pere Santamaria, Ignacio Rodríguez-Pizá, Xavier Clemente-Casares, Jun Yamanouchi, Lola Mulero-Perez, Trond Aasen, Angel Raya1 and Juan Carlos Izpisúa Belmonte. Turning Human Epidermis Into Pancreatic Endoderm. The Review of DIABETIC STUDIES 159 Vol. 7-No. 2-2010. DOI 10.1900/RDS.2010.7.158.

This pub references use of a variety of markers for showing differentiation of Human embryonic stem (hES)and induced pluripotent stem (iPS)into into pancreatic endoderm structures.

Real-time PCR and flow cytometry. A: Changes in the levels of different mRNAs in undifferentiated, and progressively differentiated, hES and iPS cells. A series of plates were cultured, as described in the materials and methods section. These were used for RNA extraction at the end of each differentiation stage, immediately prior to change in culture conditions (days 1, 3, 7, 10, and 13). B: Flow cytometric analysis of Sox17 and FoxA2 expression on cells harvested on day 3.

STEMEZ hNP1 Neural Progenitors Now Available

We are pleased to announce the introduction of our STEMEZ ^TMhNP1 Human Neural Progenitors Discovery Kit. The kit was developed by our partner, ArunA Biomedical.

Description/Data:
STEMEZ^TM hNP1 Human Progenitors are fully differentiated are derived as adherent cells from hESC WA09 line. The cells are shipped frozen in a vial with 1 x 10⁶ cells; once thawed they should be immediately plated on a Matrigel (or other suitable extracellular matrix protein)-coated dish and maintained in the accompanying serum-free medium. The neurons should be used within 14 days of thawing. These cells have the capabilities to:

• Display immunoreactive properties consistent with neural precursors and can be maintained in a proliferative state in monolayer cultures.
• Differentiate under serum-free conditions into neuronal phenotypes with functionally responsive transmitter receptors in vitro .
• Maintain multiple neural phenotypes in long term serum-free culture.
• Can be directed to alter phenotypic characteristics under different media conditions.

Applications: Gene Expression Analysis Western blotting, Flow Cytometry, Immunocytochemistry, FACS sorting, DNA Microarray, RT-PCR, Neurite Outgrowth assays, FLIPR calcium assays, cytotoxicity, and second messenger signaling.

Images: STEMEZ^TM hNP1 Human Neural Progenitor Cells are grown as monolayers (A), are karyotypically normal (B) and express NSC markers, Nestin Mouse and Sox-2 (C, D, E). Nuclei of the cells were visualized with DAPI (blue). The Sox-2 transcription factor is co-localized with the DAPI (blue) staining in the nucleus (F).

STEMEZ hNP1 FAQs

Note: STEMEZTM NP1 Progenitors and hN2 Human Neurons can be genetically manipulated with GFP reporters without altering the function and differentiation potential of these cells. In contrast to many stem cell sources, the reporter genes are not readily silence in the neural cultures. See: doi:10.1089/ten.tea.2009.0155.

DNA fingerprint cells: The loci match the DNA fingerprint pattern for the H9 (NIH designation, WA09) HESC line as published in http://stemcells.nih.gov/research/nihresearch/scunit/.

Viral tests cells: This lot was derived from the H9 hESC line that has been tested for Hepatitis B, Hepatitis C, HIV-1, HIV-2, HTLV-I/II, HSV1, HSV2, EBV, and CMV. The H9 cell line has been tested and shown to be negative. (Tests performed by GIVF Laboratories).

Related Reagents:

• STEMEZ(TM) hN2 Human Neurons Discovery Kit


• Neuron/Glial Marker Antibodies


• Neurotrophins and Growth Factor Antibodies


• Fibroblast Growth Factor Proteins


• Neurotrophins-Neuron/Glial Marker Proteins


• Stem Cell Reagents


• Transfection Reagents

STEMEZ hN2 Human Neurons-Data

Neuromics rolled out STEMEZ^TM hN2 Human Neurons Discovery Kits several months ago.

Applications for these include: cellular model studies, high content screening, developmental studies, RNAi studies and genetic manipulation.

Drilling down further, I am pleased to present Electro-physiology and related data generated by Aruna and collaborators: hN2 Cells-Electro Phys Data Supplement

hN2-Whole Cell Voltage Clamp
Figure. hN2 cells can produce inward currents that generate action potentials. (A) Isolated hN2 with significant neurite growth 1 week after plating . This cell was subjected to whole cell voltage clamp utilizing a potassium gluconate based intracellular solution. (B) Voltage gated inward and outward currents were elicited from this cell with depolarizing voltage steps. (C) Inward currents from another cell (potassium gluconate intracellular) were abolished by local application of 1 µM tetrodotoxin (red trace) while outward currents remained. Inward current recovered as TTX washed out of the region (green trace). (D) A different cell which exhibited voltage activated inward currents that inactivated in response to a 50 ms prepulse at different membrane potentials. The experiment was done 27 days after the removal of bFGF. A cesium gluconate based intracellular solution was used for this experiment to block outward potassium currents. The membrane potential for half maximal inactivation by standard Boltzman fitting (red line) was -40.1 mV with a slope of 4.7. (E) Recovery from fast inactivation utilizing a paired pulse protocol in the same cell as C. The single exponential time constant for recovery of inactivation was 1.7 ms (red line). (F) A different cell which elicited an overshooting action potential upon current injection under whole cell current clamp utilizing a potassium gluconate based intracellular solution. Inset: Response of the same cell under voltage clamp to a change in membrane potential from -80 mV to -10 mV elicited a peak current of 457 pA. Scale bars for inset: 5 ms, 0.2 nA.

Potent Stem Cell Markers

The potency of our Stem Cell Research Reagents is confirmed by the growng list of publication referencing them.

Here're the latest:

Saishu Yoshida, Takako Kato, Takao Susa, Li-yi Cai, Michie Nakayama, Yukio Kato. PROP1 coexists with SOX2 and induces PIT1-commitment cells. Biochemical and Biophysical Research Communications, Volume 385, Issue 1, 17 July 2009, Pages 11-15
... SOX2 (1:500 dilution, Neuromics, Edina, MN, USA)...

Saravanan Karumbayaram, Bennett G. Novitchb, Michaela Patterson, Joy A. Umbach, Laura Richter, Anne Lindgren, Anne E. Conway, Amander T. Clark, Steve A. Goldman, Kathrin Plath, Martina Wiedau-pazos, Harley I. Kornblum, William E. Lowry. Directed Differentiation of Human-Induced Pluripotent Stem Cells Generates Active Motor Neurons. Stem Cells Vol. 27 No. 4 April 2009, pp. 806 -811. doi:10.1002.
... mouse anti-Nestin (Neuromics)...

Patrizia Rubini, Javorina Milosevic, Johannes Engelhardt, Mahmoud Al-Khrasani, Heike Franke, Attilla Heinrich, Beata Sperlagh, Sigrid C. Schwarz, Johannes Schwarz, Wolfgang Nörenberg and Peter Illes. Increase of intracellular Ca2+ by adenine and uracil nucleotides in human midbrain-derived neuronal progenitor cells. Cell Calcium. Volume 45, Issue 5, May 2009, Pages 485-498.
...rabbit anti-P2Y2 (1:1000; Neuromics, Edina, MN, USA...

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.

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)...

Musashi Customer Data

Musashi-1

Image: Musashi-positive cells in the ventricular zone of E14.5 mouse medulla. Picture was taken at 200x magnification. The sections are 4% PFA fixed, paraffin-embedded and cut at 5 micron. Courtesy of Xi Huang, Chiang Lab, VUMC