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HGNC name: NEFMRRID: AB_2572367Immunogen: Recombinantconstruct containing the C-terminusof the human sequence (aminoacids 708-877) expressed in andpurified from E. coli.Format: Purified IgY preparation at20-30 mg/mL with 5mM NaN3Storage: Storage at 4°Crecommended. For longer termstorage make 50% wrt glycerol andput at -20°CRecommended dilutions:Use 1:10,000 for WB, 1:1,000 forICC

References:1. Harris, J., Ayyub, C. and ShawG. A molecular dissection of thecarboxyterminal tails of the majorneurofilament subunits NF-M and NF-H.J Neurosci Res 30:47-62 1991.2. Wang, Y., Zhang, J., Mori, S.and Nathans. J. Axonal Growth andGuidance Defects in Frizzled3 Knock-Out Mice: A Comparison of DiffusionTensor Magnetic Resonance Imaging,Neurofilament Staining, and GeneticallyDirected Cell Labeling. J. Neurosci.26:355-364 2006.

NF-M, chicken pAb CPCA-NF-M

Applications Host Isotype Molecular Wt. Species Cross-Reactivity

WB, IF/ICC, IHC Chicken 145-160kDaby SDS-PAGE

Hu, Rt, Ms, Bo, Po, Ho, Ck

Western blot analysis of different neuronal tissue and cell lysatesusing chicken pAb to NF-M, CPCA-NF-M, dilution 1:2,000 ingreen: [1] protein standard (red), [2] rat brain [3] rat spinal cord,[4] mouse brain, [5] mouse spinal cord, [6] NIH/3T3 cells, [7]HEK293, [8] HeLa, [9] SH-SY5Y, and [10] C6 cells. Strong bandat 145kDa corresponds to rodent NF-M, and about 160kDaband corresponds to human NF-M protein, visible in SHSY-5Yand HEK293 cells which have neuronal properties. NF-M is notexpressed in HeLa and other cell lines tested.

Immunofluorescent analysis of rat cerebellum section stained withchicken pAb to NF-M, CPCA-NF-M, dilution 1:1,000 in red, andcostained with mouse mAb to CNPase, MCA-1H10, dilution 1:500,in green. The blue is DAPI staining of nuclear DNA. Followingtranscardial perfusion of rat with 4% paraformaldehyde, brainwas post fixed for 24 hours, cut to 45μM, and free floatingsections were stained with above antibodies. The NF-M antibodylabels the axons of basket cells and other neurons, while theCNP antibody stains oligodendrocytes, cells that form the myelinsheathes around axons.

Background: Neurofilaments can be defined as the intermediate or 10nm filaments found in neuronalcells. In the electron microscope, neurofilaments appear as 10nm-diameter fibers of indeterminate lengthwhich generally have fine wispy protrusions from their sides. They are found abundantly in axons of largeprojection neurons. The possible function of neurofilaments is to provide structural support for neurons andtheir synapses, and to support the large axon diameters required for rapid conduction of impulses down axons.They are composed a mixture of subunits which usually include three neurofilament triplet proteins known asNF-L, NF-M, and NF-H. Neurofilaments may also include smaller amounts of peripherin, α-internexin, nestinand in some cases, vimentin. NF-M is the neurofilament middle or medium molecular weight polypeptide andappears on SDS-PAGE gels at 145-160kDa.

FOR RESEARCH USE ONLY. NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE.

Abbreviation Key:mAb—Monoclonal Antibody pAb—Polyclonal Antibody WB—Western Blot IF—Immunofluorescence ICC—Immunocytochemistry IHC—Immunohistochemistry E—ELISA Hu—Human Mo—Monkey Do—Dog Rt—Rat Ms—Mouse Bo—CowPo—Pig Ho—Horse Ch—Chicken Dr—D. rerio Dm—D. melanogaster Ce—C. elegans Sc—S. cerevisiae Sa—S. aureus Ec—E. coli.