Invitrogen

NEFH Monoclonal Antibody (RMdO-20)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated. View Details

1 Published Figure

9 References

View all (36) NF-H antibodies

Datasheet

Questions & Answers

Datasheet

Questions & Answers

Product Image Gallery

NEFH Antibody in Western Blot (WB) — Knockout of NEFH was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR819005_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of NEFH was performed by loading 20 µg of SH-SY5Y wild type (Lane 1), SH-SY5Y CAS9 (Lane 2), SH-SY5Y NEFH KO (Lane 3) whole cell extracts. The blot was probed with Anti-NEFH Monoclonal Antibody (RMdO-20)(Product # 13-1300) using 0.5 µg/mL dilution and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177). Loss of signal upon CRISPR mediated knockout (KO) confirms that the antibody is specific to NEFH. An uncharacterized band was observed at ~42 kDa.

NEFH Antibody in Immunohistochemistry (IHC) — Immunohistochemistry analysis of the neurofilament heavy chain showing staining in the filaments of paraffin-embedded rat 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 Immunohistochemistry (Paraffin) (IHC (P)) — 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.

NEFH Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunofluorescent analysis of the neurofilament heavy chain in frozen sections of human cortex. Tissues were probed with a neurofilament heavy chain monoclonal antibody (Product # 13-1300, red) at a dilution of 1:20, then incubated with a Alexa Fluor 594 Goat anti-Mouse IgG (H+L) Secondary Antibody. Nuclei (blue) were stained with DAPI. Data courtesy of Min Sun Kim at Seoul National University, Korea.

NEFH Antibody in Knockout — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein in NEFH KO cell line compared to control cell line using Anti-NEFH Monoclonal Antibody (RMdO-20)(Product # 13-1300). {KO}

NEFH Antibody in Relative expression — Antibody specificity was demonstrated by detection of differential basal expression of the target across cell lines tested owing to their inherent genetic constitution. Relative expression of NEFH was observed in SH-SY5Y and IMR-32 in comparison to LNCaP and MCF7 using Anti-NEFH Monoclonal Antibody (RMdO-20) (Product # 13-1300) in Western Blot. {RE}

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

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

0.5-2 µg/mL

View 1 publication 1 publication

Immunohistochemistry (IHC)

1:20

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

View 2 publications 2 publications

Immunohistochemistry (Frozen) (IHC (F))

1:20

Immunocytochemistry (ICC)

1:20

View 1 publication 1 publication

Immunofluorescence (IF)

1:20

View 2 publications 2 publications

ELISA (ELISA)

0.1-0.5 µg/mL

Immunoprecipitation (IP)

2-5 µg

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

Involved in the maintenance of neuronal caliber, neurofilaments are the intermediate filament proteins found specifically in neurons, and are composed predominantly of three major proteins called NF-L, NF-M and NF-H. Like most other intermediate filament proteins (IFPs), the expression of the different neuronal IFPs is both tissue-specific and developmentally regulated. NF-H has an important role in mature axons and is involved in amyotrophic lateral sclerosis (ALS).

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

Bioinformatics

Protein Aliases: 200 kDa neurofilament protein; NEF3; neurofilament; neurofilament 200kDa; Neurofilament heavy polypeptide; Neurofilament triplet H protein; neurofilament, heavy polypeptide; neurofilament, heavy polypeptide 200kDa; neurofilament-H; NF 200 kD; NF-H; NHC

Gene Aliases: CMT2CC; I79_012512; KIAA0845; mKIAA0845; NEFH; NF-H; NF200; NFH

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

Entrez Gene ID: (Human) 4744, (Rabbit) 100357177, (Chinese hamster) 100753127, (Rat) 24587, (Mouse) 380684

Function(s)