Overview
Product name
Description
Tested applications
Species reactivity
Immunogen
Anti-200 kD Neurofilament Heavy antibody
Chicken polyclonal to 200 kD Neurofilament Heavy
IHC-Fr, IHC-FoFr, IHC - Wholemount, ICC/IF, WB
Reacts with: Mouse, Rat, Chicken, Cat, Human, Aplysia
Full length native protein (cow intermediate filaments were prepared from spinal cords by the glycerol polymerization method of Delacourte et al., and the cytoskeletal material was dissolved in 6M urea. Individual neurofilament subunits were purified by ion exchange chromatography on
DEAE-cellulose and the NF-H containing fractions were concentrated and further purified by preparative gel electrophoresis on a Biorad Prepcell).
Properties
Form
Storage instructions
Storage buffer
Purity
Purification notes
Clonality
Isotype
Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 5mM Sodium Azide
Constituents: PBS, Whole Serum
IgY fraction
This is an IgY prep, similar to a mammalian ammonium sulphate IgG prep from serum. It is virtually pure IgY and we estimate that about 5% of material is specific to the immunogen.
Polyclonal
IgY
Applications
Our Abpromise guarantee covers the use of ab4680 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 1/10000.
IHC-FoFr 1/1000.
IHC - Wholemount Use at an assay dependent concentration.
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Application
ICC/IF
WB
Target
Function
Involvement in disease
Sequence similarities
Post-translational modifications
Abreviews Notes
Use at an assay dependent concentration. PubMed: 19944698
1/1e+006. Predicted molecular weight: 200-220 kDa.
Neurofilaments 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.
Defects 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.
Belongs to the intermediate filament family.
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.
Anti-200 kD Neurofilament Heavy antibody images
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-200 kD Neurofilament
Heavy antibody (ab4680)
This image is courtesy of an Abreview submitted by
Francoise Geffroy.
Immunohistochemical staining of 200 kDa neurofilament heavy in aplysia buccal ganglia with ab4680 at 1/1000. The sample was permeabilized with 2% Triton X100 and blocked with 1%BSA/5% goat serum for 2 hours at room temperature, before being incubated with the primary antibody for 1 hour at room temperature in 5% goat serum.
ab96949 (goat anti-chicken IgY H&L DyLight
®
594) was used as the secondary antibody at
1/200.
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Rat Neurons stained with NFH (red) and rabbit polyclonal to GFAP(green). The NF-H antibody was used at a dilution of 1/100000 and the GFAP at a dilution of 1:2,000 using our standard fixation and staining procedure
(see protocol section).
Immunocytochemistry/ Immunofluorescence -
Anti-200 kD Neurofilament Heavy antibody
(ab4680)
Immunocytochemistry/ Immunofluorescence -
Anti-200 kD Neurofilament Heavy antibody
(ab4680)
Human neuroblastoma SH-SY5Y cells stained with chicken anti-NF-H (red) and mouse monoclonal to fibrillarin 38F3 (green). Nuclear
DNA is revealed with Hoechst dye (blue). The
NF-H antibody was used at a dilution of
1/100000 and the fibrillarin monoclonal at
1/1000. Cultures were processed using our standard fixation and staining procedure (in protocol section).
Rat Neurons stained with chicken anti-NF-H
(green). The NF-H antibody was used at a dilution of 1/100000 using our standard fixation and staining procedure (in protocol section).
Immunocytochemistry/ Immunofluorescence -
Anti-200 kD Neurofilament Heavy antibody
(ab4680)
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Immunohistochemistry (Frozen sections) - 200 kD
Neurofilament Heavy antibody - Neuronal Marker
(ab4680)
This image is courtesy of an anonymous Abreview.
ab4680 at 1/10000 dilution staining neurofilament in rat skin tissue by immunohistochemistry (frozen sections).
Sections were fixed using Bouin's Fixative and permeabilized in 0.1% Triton X-100 prior to blocking in 10% serum for 1 hour at 24°C and then incubated with ab4680 for 24 hours at 4°C. A donkey polyclonal to chicken IgY, diluted 1/100 was used as the secondary antibody.
ab4680 staining 200 kD Neurofilament Heavy
(green) in murine embryoid bodies by
Immunocytochemistry/ Immunofluorescence.
Nuclei were visualized with DAPI.
Immunocytochemistry/ Immunofluorescence - 200 kD Neurofilament Heavy antibody - Neuronal
Marker (ab4680)
Image from Wirt SE et al, J Cell Biol. 2010 Nov
15;191(4):809-25. Epub 2010 Nov 8, Fig 4. DOI
10.1083/jcb.201003048
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