Product Pathways - Neuroscience
Neurofilament-H (RMdO 20) Mouse mAb #2836
PhosphoSitePlus® protein, site, and accession data: NFH
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IHC-P IF-F | H M R | Endogenous | 180-220 | Mouse IgG1 |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-F=Immunofluorescence (Frozen)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Neurofilament-H (RMdO 20) Mouse mAb detects endogenous levels of total Neurofilament-H protein. Neurofilament-H (RMdO 20) Mouse mAb has been reported to detect NFM and NFH in human samples but only NFH in mouse, rat or bovine samples (Lee, V.M. et al., 1988).
Source / Purification
Monoclonal antibody is produced by immunizing animals with rat neurofilament.
Western Blotting
Western blot analysis of extracts from mouse brain, rat brain or CAD cells, using Neurofilament-H (RMdO 20) Mouse mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human brain, using Neurofilament-H (RMdO 20) Mouse mAb.
IF-F
Confocal immunofluorescent image of mouse cerebellum labeled with Neurofilament-H (RMdO 20) Mouse mAb (green) and Calbindin Antibody #2136 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Background
The cytoskeleton consists of three types of cytosolic fibers: actin microfilaments, intermediate filaments, and microtubules. Neurofilaments are the major intermediate filaments found in neurons and consist of light (NFL), medium (NFM), and heavy (NFH) subunits (1). Similar in structure to other intermediate filament proteins, neurofilaments have a globular amino-terminal head, a central α-helical rod domain, and a carboxy-terminal tail. A heterotetrameric unit (NFL-NFM and NFL-NFH) forms a protofilament, with eight protofilaments comprising the typical 10 nm intermediate filament (2). While neurofilaments are critical for radial axon growth and determine axon caliber, microtubules are involved in axon elongation. PKA phosphorylates the head domain of NFL and NFM to inhibit neurofilament assembly (3,4). Research studies have shown neurofilament accumulations in many human neurological disorders including Parkinson's disease (in Lewy bodies along with α-synuclein), Alzheimer's disease, Charcot-Marie-Tooth disease, and Amyotrophic Lateral Sclerosis (ALS) (1).
Studies of NFH (-/-) mice suggest that NFH modulates ion channel functions in large myelinated fibers (5).
- Al-Chalabi, A. and Miller, C.C. (2003) Bioessays 25, 346-355.
- Cohlberg, J.A. et al. (1995) J. Biol. Chem. 270, 9334-9339.
- Hisanaga, S. et al. (1994) Mol. Biol. Cell 5, 161-172.
- Sihag, R.K. et al. (1999) J. Neurochem. 72, 491-499.
- Kriz, J. et al. (2000) Brain Res 885, 32-44.
Application References
- Lee, V.M. et al. (1988) Proc Natl Acad Sci U S A 85, 7384-8. Applications: Western Blotting
- Lee, V.M. et al. (1987) J Neurosci 7, 3474-88. Applications: IHC-P (paraffin) Western Blotting
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Companion Products
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- 2838 Neurofilament-M (RMO 14.9) Mouse mAb
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- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
