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Anti-Neurofilament M Purified

Neurofilaments (NFs) are a type of intermediate filament (IF) expressed almost exclusively in neuronal cells, and in those cells most prominently in large axons. NFs in most vertebrates are composed of three different polypeptide chains with different molecular weights – neurofilament heavy protein (NF-H), medium (NF-M) and light protein (NF-L), which share sequence and structural similarity in a coiled-coil core domain, but differ in the length and sequence of their N-termini and more dramatically of their C-termini which in the case of NF-M and NF-H form the flexible extensions that link NFs to each other and to other elements in the cytoplasm. The protein segment on the C-terminal side of the human NF-H rod is uniquely long (more than 600 amino acids) compared to other IF proteins and is highly charged (> 24 % Glu, > 25 % Lys), rich in proline (> 12 %) and improverished in cysteine, methionine and aromatic amino acids. Its most remarkable feature is a repetitive sequence that covers more than half its lenght and includes the sekvence motif Lys-Ser-Pro (KSP) greater than 40 times. Plasma neurofilament heavy chain level has been proposed as a marker of axonal injury and clinical use of its degeneration and loss has been suggested as a biomarker of several neurodegenerative diseases.

Product Specifications

Certification

RUO

Reactivity

Mammalian

Immunogen

Pellet of porcine brain cold stable proteins after depolymerization of microtubules.

Target Antigen

Neurofilament M

Clone

NF-09

Applications

IHC-P, WB, ICC

Concentration

1 mg/mL

Format

Purified

Buffer

Phosphate buffered saline (PBS), pH 7.4, 15 mM sodium azide

References & Citations

*Dráberová E, Sulimenko V, Vinopal S, Sulimenko T, Sládková V, D'Agostino L, Sobol M, Hozák P, Křen L, Katsetos CD, Dráber P: Differential expression of human γ -tubulin isotypes during neuronal development and oxidative stress points to a γ -tubulin-2 prosurvival function. FASEB J. 2017 May;31 (5) :1828-1846., URL: https://www.ncbi.nlm.nih.gov/pubmed/28119396 , *Draberova E, Sulimenko V, Kukharskyy V, Draber P: Monoclonal antibody NF-09 specific for neurofilament protein NF-M. Folia Biol (Praha) . 1999;45 (4) :163-5., URL: https://pubmed.ncbi.nlm.nih.gov/10732731/

Other References

*Ohara O, Gahara Y, Miyake T, Teraoka H, Kitamura T: Neurofilament deficiency in quail caused by nonsense mutation in neurofilament-L gene. J Cell Biol. 1993 Apr;121 (2) :387-95., URL: https://www.ncbi.nlm.nih.gov/pubmed/8468353?dopt=A, *Rao MV, Campbell J, Yuan A, Kumar A, Gotow T, Uchiyama Y, Nixon RA: The neurofilament middle molecular mass subunit carboxyl-terminal tail domains is essential for the radial growth and cytoskeletal architecture of axons but not for regulating neurofilament transport rate. J Cell Biol. 2003 Dec 8;163 (5) :1021-31., URL: https://www.ncbi.nlm.nih.gov/pubmed/14662746?ordin, *Deng Y, Li B, Liu F, Iqbal K, Grundke-Iqbal I, Brandt R, Gong CX: Regulation between O-GlcNAcylation and phosphorylation of neurofilament-M and their dysregulation in Alzheimer disease. FASEB J. 2008 Jan;22 (1) :138-45., URL: https://www.ncbi.nlm.nih.gov/pubmed/17687114?ordin

Storage Conditions

Store at 2-8°C. Do not freeze.

Specificity

The antibody NF-09 reacts with both phosphorylated and non-phosphorylated form of medium neurofilament protein (160 kDa intracellular antigen) of various species.

Applications Notes

Immunocytochemistry: Positive tissue: Neuro2A murine cell line, Carnoys fixative 2 x 3 min, blocking 1% glycine + 0.2% gelatin 10 min. Western blotting: Recommended dilution: 1-2 μg/ml.

Isotype

Mouse IgG2a

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