Anti-Neurofilament heavy polypeptide antibody (ab88043)

Overview

  • Product nameAnti-Neurofilament heavy polypeptide antibody
    See all Neurofilament heavy polypeptide primary antibodies
  • Description
    Rabbit polyclonal to Neurofilament heavy polypeptide
  • Tested applicationsSuitable for: IHC-Fr, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Dog
  • Immunogen

    Synthetic peptide corresponding to Human Neurofilament heavy polypeptide aa 100-200 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab105491)

  • Positive control
    • This antibody gave a positive signal in human hippocampus formalin fixed paraffin embedded tissue section.

Properties

Applications

Our Abpromise guarantee covers the use of ab88043 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • FunctionNeurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber. NF-H has an important function in mature axons that is not subserved by the two smaller NF proteins.
  • Involvement in diseaseDefects in NEFH are a cause of susceptibility to amyotrophic lateral sclerosis (ALS) [MIM:105400]. ALS is a neurodegenerative disorder affecting upper and lower motor neurons, and resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology is likely to be multifactorial, involving both genetic and environmental factors.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Post-translational
    modifications
    There are a number of repeats of the tripeptide K-S-P, NFH is phosphorylated on a number of the serines in this motif. It is thought that phosphorylation of NFH results in the formation of interfilament cross bridges that are important in the maintenance of axonal caliber.
    Phosphorylation seems to play a major role in the functioning of the larger neurofilament polypeptides (NF-M and NF-H), the levels of phosphorylation being altered developmentally and coincident with a change in the neurofilament function.
    Phosphorylated in the Head and Rod regions by the PKC kinase PKN1, leading to inhibit polymerization.
  • Information by UniProt
  • Database links
  • Alternative names
    • 200 kDa neurofilament protein antibody
    • CMT2CC antibody
    • Nefh antibody
    • Neurofilament heavy polypeptide 200kDa antibody
    • Neurofilament heavy polypeptide antibody
    • Neurofilament triplet H protein antibody
    • NF H antibody
    • NF-H antibody
    • NFH antibody
    • NFH_HUMAN antibody
    see all

Anti-Neurofilament heavy polypeptide antibody images

  • ab88043 staining Neurofilament heavy polypeptide in Zebrafish retina (neurons) tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with Triton X-100 and blocked with 5% BSA for 1 hour at 23°C. Tris-HCl was used for antigen retrieval. Samples were incubated with primary antibody (1/100) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • IHC image of Neurofilament heavy polypeptide staining in human hippocampus formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab88043, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

  • ICC/IF image of ab88043 stained PC12 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab88043, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) PC12 cells at 1µg/ml.
  • ICC/IF image of ab88043 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab88043, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Neurofilament heavy polypeptide antibody (ab88043)

ab88043 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Cryopreserved dorsal root ganglion neurons (QBMce)
Permeabilization No
Specification Cryopreserved dorsal root ganglion neurons (QBMce
Fixative paraformaldehyde with picric acid
Username

Ms. Babben Tinner

Verified customer

Submitted Nov 06 2015

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Sample Zebrafish Tissue sections (Retina, Neurons)
Specification Retina, Neurons
Permeabilization Yes - triton X
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 02 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: ciotric aid
Sample Rat Tissue sections (Spinal cord)
Specification Spinal cord
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted Jul 25 2013

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization No
Username

Dr. Sophie Pezet

Verified customer

Submitted Mar 21 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"