Anti-200 kD Neurofilament Heavy antibody (ab105453)

Overview

  • Product nameAnti-200 kD Neurofilament Heavy antibodySee all 200 kD Neurofilament Heavy primary antibodies ...
  • Description
    Rabbit polyclonal to 200 kD Neurofilament Heavy
  • Tested applicationsICC/IF, IHC-FoFr, IHC-P more details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human 200 kD Neurofilament Heavy.

  • 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 ab105453 in the following tested applications.

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

Application Notes
ICC/IF Use a concentration of 5 µg/ml.
IHC-FoFr 1/100.
IHC-P Use a concentration of 1 µg/ml.

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.
  • Target information above from: UniProt accession P12036 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • 200 kDa neurofilament protein 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-200 kD Neurofilament Heavy antibody images

  • IHC image of 200kD Neurofilament Heavy 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 ab105453, 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 ab105453 stained SKNSH 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 (ab105453, 5µg/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. This antibody also gave a positive result in 100% methanol fixed (5 min) SKNSH cells at 5µg/ml.
  • IHC-FoFr image of 200 kDa Neurofilament Heavy staining on rat hippocampus sections using ab105453 (/100).The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the spinal cord were cryoprotected in sucrose 30% overnight. Spinal cords were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

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References for Anti-200 kD Neurofilament Heavy antibody (ab105453)

ab105453 has not yet been referenced specifically in any publications.

Product Wall

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: Citric acid
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 Feb 14 2012

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