Schwann Cell Data

Quality Counts
Though rare, we occasionally have clients challenge the phenotype of our 21-CFR compliant human primary cells. This challenge is usually a result of  certain markers not being expressed when they run immuno-fluorescence assays.

When this happens, we too test the cells in question using the antibodies against the markers in question. We have a world class team doing immuno-fluorescence assays.

Recently a customer claimed our schwann cells were negative for GFAP and P0. Here're our testing results.

For P0 (CH23009) we used green fluorescence +DAPI.

For GFAP (RA22101) we used red fluorescence + DAPI (blue) counterstain

We take these challenges for seriously and will always test the same lot of cells internally to make sure they are "walking and talking" as advertised.

Neuron-Glial Markers

Featuring Neurofilament Antibodies
We have the honor of our Neuron-Glial Markers being referenced in many publications. Here we wanted to feature some recent data from one of our Neurofilament Antibodies.

Fig. Merkel cells in grafted ESS are associated with neurons expression neurofilament heavy (NF-M) by eight weeks after grafting. Immunochemistry with antibodies against NF-H (red) and KRT20 (green) was used to localize neurons and Merkel cells, respectively, in ESS after grafting to mice. Nuclei were counterstained with DAPI (blue; B, D, F, H). Shown are cross sections of ESS at 4 weeks (A-B), 8 weeks (C-D), 12 weeks (E-F), and 14 weeks (G-H) after grafting; each row contains images of the same section. Scale bars in A is same for all images. White arrows indicate examples of NF-H-positive nerves associated with or in proximity to Merkel cells; yellow arrows indicate NF-H-positive nerves not associated with Merkel cells. https://doi.org/10.1371/journal.pone.0213325.g009

Remember we offer 100% refunds should you not be delighted with the performance of our antibodies.

Cell to Cell Signaling and Depression

ATP and Your Brain
A recent article in Molecular Psychiatrydoi:10.1038/mp.2017.229 elucidates the role of ATP and Neuron-Glial interactions in depression.

This study also features the use of our excellent GFAP markers

Abstract: Extracellular ATP is a widespread cell-to-cell signaling molecule in the brain, where it functions as a neuromodulator by activating glia and neurons. Although ATP exerts multiple effects on synaptic plasticity and neuro-glia interactions, as well as in mood disorders, the source and regulation of ATP release remain to be elaborated. Here, we define Calhm2 as an ATP-releasing channel protein based on in vitro and in vivo models. Conventional knockout and conditional astrocyte knockout of Calhm2 both lead to significantly reduced ATP concentrations, loss of hippocampal spine number, neural dysfunction and depression-like behaviors in mice, which can be significantly rescued by ATP replenishment. Our findings identify Calhm2 as a critical ATP-releasing channel that modulates neural activity and as a potential risk factor of depression.

Adult Primary Neurons

Build Yourself a Stock. We now have adult primary neurons that can be passaged up to 5 times! You can now build a stock of 20-30 million cells.

Human primary neurons are isolated from brain cortical tissue. We highly recommend culturing in our human neuron growth media (cat#HNM001).
Check out our many Primary Neuron and Astrocyte options.

ECS Derived hN2 Human Primary Neurons grown in culture for 4 days and stained with our chicken polyclonal to Neurofilament light or low molecular weight chain NF-L, a marker of neurons. Many of the differentiating cells show strong cytoplasmic and clearly fibrillar staining for NF-L. Blue stain is DAPI and reveals cell nuclei of some nonneuronal cells in these cultures.
Check out related Markers!

alpha-II-spectrin/alpha-Fodrin	MO22110	Mouse IgG	H; M; R	IF; WB	100 ul	$245
Calcineurin A	MO15055	Mouse IgG	H; M; R	WB	100 ug	$255
Doublecortin/DCX	MO22113	Mouse IgG	B; H; M; P; R	IF; WB	100 ul	$245
FOX3	RA22113	Chicken IgY	H; M; R	IF; IHC; WB	100 ul	$245
MAP2 (Microtubule assoc. protein 2)	MO30000	Mouse IgG	H; M; P	IHC	50 ul	$365
Neurofilament NF-M	RA22105	Rabbit IgG	Ca; Ch; H; M; R	ICC; IHC; WB	100 ul	$275
Peripherin	CH22111	Chicken IgY	Ca; H; M; R	IHC	100 ul	$245
alpha Synuclein	MO22111	Mouse IgG	H; M; R	IF; WB	100 ul	$245
Tau /MAPT+	CH22113	Rabbit IgG	H; R	IF; WB	100 ul	$245

Neuron-Glial Antibodies Hit the Mark!

Neuromics' Neuron-Glial Markers offerings include:

Glial-Astrocyte Markers	Neural Progenitor Markers
Neuron/Synapse Markers	Oligodendrocyte, Oligodendroglial Oligodendrocyte Lineage Markers
Schwann Cell or PNS Neuronal Markers -Peripheral Nervous System (PNS) Related

They are widely used and frequently published. Customers have also shared data which is included in product descriptions. Here is a listing of the most recent pubs:
Wiebke Kallenborn-Gerhardt, Katrin Schröder, Domenico Del Turco, Ruirui Lu, Katharina Kynast, Judith Kosowski, Ellen Niederberger, Ajay M. Shah, Ralf P. Brandes, Gerd Geisslinger, and Achim Schmidtko. NADPH Oxidase-4 Maintains Neuropathic Pain after Peripheral Nerve Injury. The Journal of Neuroscience, 25 July 2012, 32(30): 10136-10145; doi: 10.1523/​JNEUROSCI.6227-11.2012.

Images: MPZ and PMP22 expression in the sciatic nerve of WT and Nox4−/− mice after SNI. A, Western blot analysis of the myelin-specific proteins MPZ and PMP22 in the day 14 SNI sciatic nerve (proximal nerve stump) and the uninjured control sciatic nerve. GAPDH was used as loading control. Note that MPZ and PMP22 protein expression is significantly decreased after SNI in WT mice but not in Nox4−/− mice. n = 3 mice per group. Data are presented as mean ± SEM (*p < 0.05). B, Immunostaining of the day 14 SNI sciatic nerve shows increased MPZ immunoreactivity in Nox4−/− mice compared with WT mice, whereas immunoreactivity of the neuronal marker NF200 is similar in both genotypes. Scale bar, 10 μm.

Maria Maddalena Valente, Valeria Bortolotto, Bruna Cuccurazzu, Federica Ubezio, Vasco Meneghini, Maria Teresa Francese, Pier Luigi Canonico, Mariagrazia Grilli.Alpha2delta ligands act as positive modulators of adult hippocampal neurogenesis andprevent depressive-like behavior induced by chronic restraint stress. Molecular Pharmacology Fast Forward Published on May 9, 2012 as doi:10.1124/mol.112.077636...chicken anti-nestin polyclonal (1:4,000, Neuromics, Edina, Minnesota)...

Frances Y. Cheng, Xi Huang, Anuraag Sarangi, Tatiana Ketova, Michael K. Cooper, Ying Litingtung, Chin Chiang. Widespread Contribution of Gdf7 Lineage to Cerebellar Cell Types and Implications for Hedgehog-Driven Medulloblastoma Formation. PLoS ONE 7(4): e35541. doi:10.1371/journal.pone.0035541...rabbit anti-GFAP (Neuromics, 1:500), mouse anti-GFAP (Neuromics, 1:200)...

Juan M Jimenez-Andrade and Patrick W Mantyh. Sensory and sympathetic nerve fibers undergo sprouting and neuroma formation in the painful arthritic joint of geriatric mice. Arthritis Research & Therapy 2012, 14:R101...to label primary afferent sensory nerve fibers, an antibody against neurofilament 200 kDa (NF200, chicken anti neurofilament 200 kDa; NF200, 1:5000; Neuromics; catalog #CH22104)...

I will be posting new developments.

Early Diagnosis of Diabetic Retinopathy

The earlier the diagnosis the better the outcome. This is especially true with autoimmune diseases like Diabetic Retinopathy (DR). DR is the leading cause of blindness among persons of working age in the industrialized world. Here I feature a publication that shows axoglial alterations at the distal portion of the optic nerve could be the first structural change in the diabetic visual pathway. This could prove good news for discovering better therapies thus preventing blindness: Diego C. Fernandez, Laura A. Pasquini, Damián Dorfman, Hernán J. Aldana Marcos, Ruth E. Rosenstein. Early Distal Axonopathy of the Visual Pathway in Experimental Diabetes. doi:10.1016/j.ajpath.2011.09.018

Oligodendrocytes are responsible for insulating axons. Disruptions in the formation of oligodendrocytes could initiate the domino effect that leads to decreasing and eventual total loss of vision. The authors, for example, discovered that in diabetic rats, oligodendrocyte lineage (OL) cells showed hypertrophic somas and a high number of processes.

Images/Data: OL linage evaluation. Immature OL (O1+ cells) and OL precursor (PDGFR-α+ cells) were evaluated by immunostaining and measured as optical density (OD) per section. In the distal ON from animals that were diabetic for 6 weeks, significantly increased O1 and PDGFR-α immunostaining was observed, with the presence of disorganized and hypertrophic cells. Data are mean ± SEM (n = 5 animals per group); *P < 0.01 versus age-matched controls, by Student′s t-test. Scale bar = 50 μm.

At the ultrastructural level, alterations and loss of larger axons were observed in the distal ON from animals that were diabetic for 6 weeks. In these fibers, myelin was highly disorganized, and frequent lamellar membranous bodies were observed.

I will track new develops in this research and post relevant results here.

GFAP and Mouse Myenteric Plexus

Our Neuronal-Glial Markers are important tools for our customers investigating expression in the CNS and PNS. I have posted images showing staining of mouse retinal astrocytes and in the ventral horn, funiculus of adult rat spinal cord and mouse medulloblastoma stem cells using our GFAP antibodies.

I wanted to share an excellent image generated by Dr. Kate Ellacott's lab at Vanderbilt University.

GFAP (Chicken-Cat#: CH22102) staining in of enteric glia in the myenteric plexus of the mouse gastrointestinal tract. Staining was performed in methanol/acetone fixed frozen sections using 1:1000 dilution of the antibody followed by 1:500 anti-chicken Alexa 594.

Related Reagents:

• GFAP (Chicken-Cat#: CH23011)
• GFAP (Mouse-Cat#; MO15052)
• GFAP (Rabbit-Cat#:RA19063)
• Neuronal-Glial Markers -Astrocytes, Glia, Microglia, Olidogodendrocytes, Progenitors and Schwann Cell.

Staining Neuron-Glial Cultures-Related Markers

I have been receiving a growing number of requests for best techniques related to staining cultures of primary neurons and glia. I wanted to share this short, step by step protocol.

These requests are often catalyzed by a search of our growing Neuron/Glial Markers catalog. The objective being to find the right markers for a particular assay. I wanted to share examples of the potency of several Neurofilament or NF markers for labeling neurons:

1. Neurofilament NF-L-Mouse Monoclonal Antibody (Clone: DA2) and Neurofilament alpha-internexin/NF66-Whole Serum-Rabbit Antibody

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-L. Protocols on data-sheet.

2. Neurofilament NF-H, phosphylated-Mouse Monoclonal and Neurofilament NF-L-Purified Chicken Polyclonal.

Image: View of mixed neuron/glial cultures stained with chicken polyclonal NF-L (red) and phosphorylated NF-H The NF-L protein is assembled into neurofilaments which are found throughout the axons, dendrites and perikarya of these cells. In contrast the phosphorylated NF-H has a much rmore restricted expression pattern, being found only in developed axonal neurofilaments. Since both proteins are found in neurofilaments, the red and green patterns overlap, so that neurofilaments containing NF-L and phosphorylated NF-H appear yellowish. In contrast neurofilaments containing only NF-L appear red. Protocol on datasheet.

Neurofilament Markers