Biolegend, 801707, HRP anti-Neurofilament H (NF-H), Nonphosphorylated Antibody, 25μg

Neurofilaments (NF) are approximately 10 nanometer intermediate filaments found in neurons. They are a major component of the neuronal cytoskeleton, and function primarily to provide structural support for the axon and to regulate the axon diameter. There are three major NF subunits, and the names given to these subunits are based upon the apparent molecular mass of the mammalian subunits on SDS-PAGE. The light or lowest NF (NF-L) runs at 68-70 kD. The medium or middle NF (NF-M) runs at about 145-160 kD, and the heavy or highest NF (NF-H) runs at 200-220 kD. However, the actual molecular weight of these proteins is considerably lower due to the highly charged C-terminal regions of the molecules. The level of NF gene expression correlates with the axonal diameter, which controls how fast electrical signals travel down the axon. Mutant mice with NF abnormalities have phenotypes resembling amyotrophic lateral sclerosis. NF immunostaining is common in diagnostic neuropathology. It is useful for differentiating neurons (positive for NF) from the glia (negative for NF).
25μg
Verified Reactivity: Human, Mouse, Rat
Antibody Type: Monoclonal
Host Species: Mouse
Formulation: This antibody is provided in 50% glycerol in aqueous buffered solutions with preservatives.
Preparation: The antibody was purified by affinity chromatography and conjugated with HRP under optimal conditions.
Concentration: 0.5 mg/ml
Storage & Handling: Upon receipt, the antibody solution should be stored undiluted at -20°C, and protected from prolonged exposure to light.
Application: IHC-P - Quality tested WB - Verified
Recommended Usage: Each lot of this antibody is quality control tested by immunohistochemistry staining. For immunohistochemisty of paraffin-embedded tissue, a concentration range of 5.0 to 10.0 µg/ml is suggested. For Western Blot, a concentration range of 0.5 to 1.0 µg/ml is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes: Additional reported applications (for the relevant formats) include Western blotting6, immunohistochemistry4,5, immunocytochemistry1,2,3, 7, array tomography8. Cross-reactivity to monkey tissue has been Reported in the literature, not verified in house4. This antibody reacts with a nonphosphorylated epitope in neurofilament H of most mammalian species. The reaction is masked when the epitope is phosphorylated. The staining of isolated neurofilament preparations is greatly intensified upon dephosphorylation. Immunocytochemically, SMI 32 visualizes neuronal cell bodies, dendrites, and some thick axons in the central and peripheral nervous systems. However, thin axons are not revealed. Other cells and tissues are unreactive. The antibody distinguishes three subdivisions of the macaque precentral motor cortex. The greater size of the left versus the right superior temporal lobe was found to be due to increased axonal myelination and not due to increased number of glial cells or SMI 32-enumerated neurons, suggesting that the specialization for language in the left temporal lobe is related to increased speed of signal transmission. In cultures of murine cortex, SMI 32 labels a neuronal population with enhanced vulnerability to kainate toxicity most of which are GABAergic and reveal kainate-activated Ca2+ uptake.
Application References(PubMed link indicates BioLegend citation): Chang Q, Martin LJ. 2011. J. Neurosci., 31:2815-27. (ICC) PubMed Stevens HE, et al. 2010. J. Neurosci. 30:5590-602. (ICC) PubMed Kiryu-Seo S, et al. 2010. J. Neurosci. 30:6658-66. (ICC) PubMed Redondo J, et al. 2015. Brain Pathol. 25(6):692. (IHC-P) PubMed Feng L, et al. 2017. eNeuro. 4(1): 0331-16.2016. (IHC-P) PubMed Feng L, et al. 2014. Invest Ophthalmol Vis Sci. 54(2): 1106:1117. (IHC-P) PubMed Theotokis, et al. 2016. J. Neuroinflammation 13(1):265 (IHC-P) Bennett, et al. 2015. J. Neurosci. Methods 245:25-36 (Array Tomography) Petzold A, et al. 2011. Brain 134:464. (WB) PubMed
RRID: AB_2721607 (BioLegend Cat. No. 801707) AB_2721608 (BioLegend Cat. No. 801708)
Structure: Neurofilament H has an apparent molecular mass of 200-220 kD.
Distribution: Tissue Distribution: CNS, peripheral nerves and glandular cells of the prostate Cellular Distribution: cytoskeleton, nucleus, cytosol, and mitochondrion
Function: NF-H Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter. Phosphorylation of NF-H results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
Ligand/Receptor: Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation result in changes to the neurofilament function.
Cell Type: Mature Neurons
Biology Area: Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family: Intermediate Filaments, Phospho-Proteins
Antigen References: 1. Turner M, et al. 2015. J Neuroimmunol. 285:4. PubMed 2. Pagliarini V, et al. 2015. J. Cell Biol. 211:77. PubMed 3. Petzold A, et al. 2011. Brain. 134. (WB) PubMed 4. Yuan A, et al. 2016. Brain Res Bull. 126(3):334. 5. Parlakian A, et al. 2016. Rev Neurol. 172(10):607. 6. Li D, et al. 2016. Front Aging Neurosci. 8:290. 7. Costa J, et al. 2016. Clin Chim Acta. 455:7. 8. Lad SP, et al. 2010. J Stroke Cerebrovasc Dis. 21(1):30.
Gene ID: 4744
UniProt: View information about Neurofilament H on UniProt.org
Clone: SMI 32
Regulatory Status: RUO
Other Names: Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Isotype: Mouse IgG1