Biolegend, 835609, Alexa Fluor® 594 anti-Neurofilament H & M (NF-H/NF-M), Hypophosphorylated Antibody, 25μg

Neurofilaments (NFs) 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 neurofilament (NF-L) runs at 68-70 kD. The medium or middle (NF-M) runs at about 145-160 kD, and the heavy or highest (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: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions.
Concentration: 0.5 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: IHC-P - Quality tested ICC - Verified
Recommended Usage: Each lot of this antibody is quality control tested by formalin-fixed paraffin-embedded immunohistochemical staining. For immunohistochemistry, a concentration of 5.0 µg/mL is suggested. For immunocytochemistry, a concentration range of 5.0 - 10 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application. * Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses
Application Notes: Additional reported applications (for the relevant formats of this clone): include ELISA Capture1-3, immunohistochemical staining on frozen tissue sections, immunofluorescence staining, and spatial biology (IBEX)6,7. Clone SMI 35 reacts with highly phosphorylated neurofilaments, as well as with low degrees of phosphorylation. It primarily reacts with neurofilament H and with neurofilament M to a lesser extent. Notes: On two dimensional gels, this antibody detects a band extending from the phosphorylated neurofilament position at 200 kD (pI 5.1) toward the non-phosphorylated position at 170 kD (pI 6.2).
Application References(PubMed link indicates BioLegend citation): Petzold A. 2013. J. Neuroimmunol. 262:(1-10). (ELISA) Lu CH, et al. 2012. PLoS One. 7:e40998. (ELISA) Steinacker P, et al. 2011. PLoS One. 8:e23600. (ELISA) Poltorak M, et al. 1993. J. Neurosci. 13:2217. (IHC-F) Petzold A, et al. 2011. Brain. 134:464. (WB) PubMed Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
RRID: AB_2687301 (BioLegend Cat. No. 835609) AB_2687299 (BioLegend Cat. No. 835610)
Structure: 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.
Distribution: Tissue Distribution: CNS, peripheral nerves and glandular cells of the prostate Cellular Distribution: Cytoskeleton, nucleus, cytosol, and mitochondrion
Function: Neurofilaments are the major components of the neuronal cytoskeleton. They provide axonal support and regulate axon diameter.
Interaction: Cell bodies and dendrites are generally unstained. Other cells and tissues are unreactive except for peripheral axons.
Cell Type: Mature Neurons
Biology Area: Cell Biology, Neuroscience, Neuroscience Cell Markers
Molecular Family: Intermediate Filaments, Phospho-Proteins
Antigen References: Petzold A. 2012. Mult Scler Int. 2012:217802. PubMed Gresle MM, et al. 2011. Mult Scler Int. 2011: 315406. PubMed
Gene ID: 4744
UniProt: View information about Neurofilament HM NF-H NF-M Phospho on UniProt.org
Clone: SMI 35
Regulatory Status: RUO
Other Names: Neurofilament heavy polypeptide, NF-H, 200 kD neurofilament protein, neurofilament triplet H protein
Isotype: Mouse IgG1, κ