NFM protein was purified from bovine spinal cords following the method of Delacourte et al. ["Study of the 10-nm-filament fraction isolated during the standard microtubule prepartion" (1980). Biochem. J. 191(2): 543-546], followed by Prepcell purification (Bio-Rad). Antibodies were prepared by injecting purified NFM protein into laying hens and purifying the IgY fraction from eggs collected from hyper-immunized hens. After several boosts, hens were co-boosted with recombinant NFM protein expressed in bacteria. These antibodies have been validated with human, mouse and rat tissues.
Immunocytochemical staining of NF-M immunoreactivity (green) and Glial Fibrillary Acidic Protein (GFAP, red) in a culture of mouse cortical neurons. The chicken anti-NFM immunostaining was performed using a 1:2000 dilution, and the GFAP immunostaining was performed using a 1:5000 dilution. Photomicrographs from Dr. Gerry Shaw, EnCor Biotechnology, Inc.
Human Neurofilament Medium Chain (NFM) is a 102,448 dalton protein (916 amino acids) expressed in CNS and PNS neurons. This cytoskeletal protein is normally phosphorylated in vivo and its extent of phosphorylation correlates with different states of axonal growth and stabilization. These observations have led to the hypothesis that the NFM protein cross-links adjacent neurofilament strands and contributes to the structural integrity of the axon.
IgY Fraction
2 mg/mL
Polyclonal
IgY
ICC, IHC, WB
Chicken
NEFM NEF3 NFM
150 kDa
Recombinant construct containing the C-terminus of the human sequence expressed in Escherichia coli
Human, Mouse, Rat
AB_2313554
Store at 4°C in the dark. Under these conditions, the antibodies should have a shelf life of at least twelve months, provided they remain sterile. For longer term storage, aliquot and freeze to avoid freeze-thaw of the antibody.
Liquid
NFM protein was purified from bovine spinal cords following the method of Delacourte et al. ["Study of the 10-nm-filament fraction isolated during the standard microtubule prepartion" (1980). Biochem. J. 191(2): 543-546], followed by Prepcell purification (Bio-Rad). Antibodies were prepared by injecting purified NFM protein into laying hens. After repeated injections into the hens, immune eggs were collected, and the IgY fractions were purified from the yolks. Antibodies were diluted in the buffer described above, and then filter-sterilized.
Phosphate-buffered (10 mM) isotonic (0.9%, w/v) saline ("PBS" pH 7.2) with bovine serum albumin (BSA, 0.5%) added to prevent absorption to the plastic and sodium azide (0.02%, w/v) added as a preservative
Quality assurance analysis was performed using immunohistochemistry (at a dilution of 1:5000) using fluorescein-labeled goat anti-chicken IgY (1:500 dilution, Aves Labs Cat.# F-1005) as the secondary reagent.
These antibodies are to be used as research laboratory reagents and are not for use as diagnostic or therapeutic reagents in humans.
United States
12 months from receipt of product
Ice packs
Neurofilament medium polypeptide (NF-M) (160 kDa neurofilament protein) (Neurofilament 3) (Neurofilament triplet M protein)
UniProt (Immunogen Species): P07197
Product Specific References for Applications and Species
Molina-Holgado, E., et al. 2021. Cannabinoid Receptor 1 associates to different molecular complexes during GABAergic neuron maturation. Journal of Neurochemistry, 640-656.
Bin, J.M., et al. 2012. Oligodendrocyte precursor cell transplantation into organotypic cerebellar shiverer slices: a model to study myelination and myelin maintenance. PLoS One, e41237.
Wang, S., et al. 2020. Identification of Medium-Length Antineurofilament Autoantibodies in Patients with Anti-N-Methyl-D-Aspartate Receptor Encephalitis. Journal of Clinical Neurology, 470-479.
Arevalo-Martin, A., et al. 2018. Elevated Autoantibodies in Subacute Human Spinal Cord Injury Are Naturally Occurring Antibodies. Frontiers in Immunology, 2365.
Gluck, M.J., et al. 2018. Detecting structural and inflammatory response after in vivo stretch injury in the rat median nerve via second harmonic generation. Journal of Neuroscience Methods, 68-80.
Bulk Order Anti-NFM (Neurofilament Medium Chain) Antibody
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