EnCor Biotechnology

Chicken Polyclonal Antibody to Neurofilament NF-M (Nfm, NEFM) Cat# CPCA-NF-M

$250.00
Description

      The CPCA-NF-M antibody was made against a recombinant fusion protein containing the C-terminal segment of rat NF-M, a region conserved across species boundaries and containing some interesting peptide repeats of currently unknown function (8). The CPCA-NF-M antibody is very similar in properties to a rabbit polyclonal the production and characterization of which were described in reference 7. As shown here, the antibody works well for western blotting, IF, ICC and IHC. Also available from EnCor is a rabbit polyclonal and a widely used mouse monoclonal antibody to the same immunogen RPCA-NF-M, and MCA-3H11. All three antibodies works on a variety of species and are clean and specific on western blots, cell and tissue staining.
Amount: 100µL of serum
Amount: 100µL of serum
Immunofluorescent analysis of rat cerbellum section stained with chicken pAb to NF-M, CPCA-NF-M, dilution 1:1,000 in red, and costained with mouse mAb to CNP, MCA-1H10, dilution 1:500 in green. The blue is DAPI staining of nuclear DNA. Following transcardial perfusion of rat with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free-floating sections were stained with above antibodies. NF-M antibody labels network of axons of basket neurons and other neuronal cells. The CNP antibody stains the oligodendrocytes, cells that create a myelin sheath around axons.
Western blot analysis of different neuronal tissue and cell lysates using chicken pAb to NF-M, CPCA-NF-M, dilution 1:2,000 in green: [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 band at 150 kDa corresponds to rodent NF-M , and about 170 kDa band corresponds to human NF-M protein.

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Immunogen: Recombinant fusion protein containing the extreme C-terminal segment of human NF-M, amino acids 741-916
HGNC Name: NEFM
UniProt: P07197
Molecular Weight: 145-160kDa
Host: Chicken
RRID: AB_2572367
Format: Concentrated IgY preparation in PBS plus 0.02% NaN3
Applications: WB, IF, ICC, IHC
Recommended Dilutions: WB: 1:2,000-5,000. IF/ICC & IHC: 1:1,000-1:2,000.
Storage: Storage for short term at 4°C recommended, for longer term at -20°C, minimize freeze/thaw cycles

Neurofilaments are the 10nm or intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. NF-M is the neurofilament middle or medium molecular weight polypeptide and runs on SDS-PAGE gels at 145-160kDa, with some species variability, though the real molecular weight is ~105kDa. The major function of neurofilaments is likely to control the diameter of large axons (1). Antibodies to NF-M such as RPCA-NF-M are useful for identifying neuronal cells and their processes in tissue sections and in cell culture. NF-M antibodies can also be useful to visualize neurofilament rich accumulations seen in many neurological diseases, such as Amyotrophic Lateral Sclerosis (a.k.a. Lou Gehrig’s disease) and Alzheimer’s disease (2-4).Much recent evidence has suggested that the detection of NF-L and NF-H in blood and CSF might be a useful prognostic or diagnostic biomarkers of neuronal damage and degeneration associated with a variety of CNS pathologies (5,6). The potential utility of NF-M in this fashion has not to date been examined.



Chromogenic Immunostaining of a formalin fixed paraffin embedded human cerebellum with chicken pAb to NF-M, CPCA-NF-M, dilution 1:2,000, detected with DAB (brown) following the ABC method with citra buffer retrieval. Hematoxylin (blue) was used as the counterstain. The NF-M antibody detects perikarya and dendrites of neuronal Purkinje cells, and is strongly expressed in the axons of basket and other kinds of neuron. Mouse select image for larger view.

1. Hoffman et al. Neurofilament gene expression:a major determinant of axonal caliber. PNAS 84:3472-6 (1987).
2. Perrot R, et al. Review of the Multiple Aspects of Neurofilament Functions, and their Possible Contribution to Neurodegeneration. Mol. Neurobiol. 38:27-65 (2008).
3. Lépinoux-Chambaud C. Eyer J. Review on intermediate filaments of the nervous system and their pathological alterations. Histochem. Cell Biol. 140:13-22 (2013).
4. Liu Q. et al. Neurofilamentopathy in Neurodegenerative Diseases. Open Neurol. J. 5:58–62 (2011).
5. Bacioglu M, et al. Neurofilament light chain in blood and CSF as marker of disease progression in mouse models and in neurodegenerative diseases. Neuron 91:56-66 (2016).
6. Shaw G. The use and potential of pNF-H as a general blood biomarker of axonal loss: an immediate application for CNS injury. in Brain Neurotrauma: Molecular, Neuropsychological, and Rehabilitation Aspects. CRC Press/Taylor & Francis Chapter 21 (2015).
7. Harris J, Ayyub C. and Shaw G. A molecular dissection of the carboxyterminal tails of the major neurofilament subunits NF-M and NF-H. J. Neurosci. Res. 30:47-62 (1991).
8. Shaw G. Identification of previously unrecognized sequence motifs at the extreme carboxyterminus of the neurofilament subunit NF-M. BBRC 14;162:294-9 (1989).

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