Invitrogen

NEFH Monoclonal Antibody (RMdO-20)

Product Image Gallery

NEFH Antibody — Published figure using NEFH monoclonal antibody (Product # 13-1300)

NEFH Antibody in Immunohistochemistry (IHC) — Immunohistochemistry analysis of the neurofilament heavy chain showing staining in the filaments of paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Neurofilament H chain monoclonal antibody (Product # 13-1300) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

NEFH Antibody in Western Blot (WB) — Western blot analysis of neurofilament heavy chain was performed by loading 25 µg of mouse brain tissue lysate per well onto a polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked. NF-H was detected at ~180-220 kD using a neurofilament heavy chain antibody (Product # 13-1300) at a dilution of 2 µg/mL in blocking buffer overnight at 4C, followed by a HRP-labeled secondary antibody for 1 hour at room temperature and detection with a chemiluminescent substrate.

NEFH Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of the neurofilament heavy chain in paraffin-embedded human brain tissue (right) compared to a negative control without primary antibody (left). Tissue sections were deparaffinized with xylene, and rehydrated with ethanol. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and microwaved for 8-15 min. Following antigen retrieval, tissues were washed with water and PBS, and then blocked in 0.3% BSA for 30 min at room temperature. Tissues were then probed with a neurofilament heavy chain monoclonal antibody (Product # 13-1300) in 0.3% BSA at a dilution of 1:20 for 1 hour at 37°C. Tissues were then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, DyLight 488 conjugate for 1 hour at 37°C (green). Nuclei (blue) were stained with DAPI. Images were taken at 40X magnification.

Product Details

Tested Applications

Dilution

ELISA (ELISA)

0.1-0.5 µg/mL

Immunocytochemistry (ICC)

1:20

Immunofluorescence (IF)

1:20

Immunohistochemistry (IHC)

1:20

Immunoprecipitation (IP)

2-5 µg

Western Blot (WB)

0.5-2 µg/mL

Published Applications

Immunohistochemistry (Paraffin) (IHC (P))

See 2 publications below

Immunohistochemistry (IHC)

See 2 publications below

Western Blot (WB)

See 1 publication below

Immunofluorescence (IF)

See 2 publications below

Immunocytochemistry (ICC)

See 1 publication below

Product Specifications

Species Reactivity

Bovine, Hamster, Human, Mollusc, Mouse, Rabbit, Rat

Published species

Human, Mouse, Rat

Host / Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

RMdO-20

Immunogen

Adult Rat neurofilaments

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

purified

Storage buffer

PBS

Contains

0.1% sodium azide

Storage conditions

-20°C

RRID

AB_2532999

Product Specific Information

This antibody reacts with 200 kDa subunit protein of human neurofilament. It specifically recognizes a nonphosphorylated epitope in the tail domain of NF-H. Dephosphorylation of NF-H will increase immunoreactivity with this antibody.

This antibody is suitable for immunohistochemical staining of Bouin's and alcohol-fixed paraffin-embedded or frozen tissue sections. To stain, incubate 30-60 minutes at room temperature or overnight at 4°C.

Target Information

Neurofilaments are the intermediate (10-12 nm) filament proteins (IFPs) located specifically in neurons, and are composed predominantly of three classes/subunits of neurofilaments: NF-L (low molecular weight; 68 kDa), NF-M (medium molecular weight; 160 kDa), and NF-H (high molecular weight; 200 kDa), with some variation in different species. The neurofilaments are long helical proteins involved in the maintenance of neuronal caliber, which polymerize to form a rigid cytoskeleton in the neuron. Like most other IFPs, the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. This polymerized network is composed of all three filaments, and the stoichiometry of association varies during development. Neurofilaments are posttranslationally modified both by phosphorylation and glycosylation. Like other intermediate filament proteins, phosphorylation likely mediates neurofilament dynamics. However, how O-linked N-acetylglucosamine residues function to modify neurofilament function remains unclear. The NF-H polyclonal antibody can be used to identify neurons and their processes in tissue sections and in tissue culture and the antibody can also be used to study microfilament accumulations seen in many neurological diseases, such as Lou Gehrig's disease or Alzheimer's disease.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: 200 kDa neurofilament protein; NEF3; NEFM; neurofilament 200kDa; Neurofilament heavy polypeptide; neurofilament medium polypeptide; neurofilament medium polypeptide 150kDa; Neurofilament triplet H protein; neurofilament triplet M protein; neurofilament, heavy polypeptide 200kDa; neurofilament, heavy polypeptide 200kDa-like; neurofilament-3, NFH, NF-H, NHC, 200 kDa neurofilament protein, KIAA0845, Neurofilament heavy polypeptide, Neurofilament triplet H protein, CMT1F, CMT2E, Neurofilament heavy; NF-H; NFM

Gene Aliases: CMT2CC; KIAA0845; mKIAA0845; NEFH; NF-H; NF200; NFH

UniProt ID: (Human) P12036, (Mouse) P19246

Entrez Gene ID: (Rabbit) 100357177, (Human) 4744, (Rat) 24587, (Mouse) 380684

Function(s)