Anti-Neurofilament heavy polypeptide antibody (ab4680)
- Datasheet
- References (29)
- Protocols
Overview
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Product nameAnti-Neurofilament heavy polypeptide antibody
See all Neurofilament heavy polypeptide primary antibodies -
DescriptionChicken polyclonal to Neurofilament heavy polypeptide
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Host speciesChicken
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Tested applicationsSuitable for: IHC-Fr, IHC-FoFr, IHC - Wholemount, ICC/IF, IHC-FrFl, WBmore details
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Species reactivityReacts with: Mouse, Rat, Cow, Human, Aplysia
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Immunogen
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).
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Positive control
- IHC-FrFl: Rat cerebellum tissue. WB: Mouse, rat and cow spinal cord lysate. IHC-PerFo: Aplysia buccal ganglia. ICC/IF: Mixed neuron/glial cultures. SH-SY5Y cells. Rat neurons. Murine embryoid bodies. IHC-Fr: Rat skin tissue.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Storage bufferPreservative: 0.032% Sodium azide
Constituents: PBS, Whole serum -
Concentration information loading... -
PurityIgY fraction
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Purification notesThis 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.
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ClonalityPolyclonal
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IsotypeIgY
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Research areas
Associated products
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Related Products
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. | |
| ICC/IF | Use at an assay dependent concentration. PubMed: 19944698 | |
| IHC-FrFl | Use at an assay dependent concentration. | |
| WB | 1/1e+006. Predicted molecular weight: 200-220 kDa. |
Target
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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.
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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.
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Sequence similaritiesBelongs to the intermediate filament family.
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Post-translational
modificationsThere 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
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Database links
- Entrez Gene: 528842 Cow
- Entrez Gene: 4744 Human
- Entrez Gene: 380684 Mouse
- Entrez Gene: 24587 Rat
- Omim: 162230 Human
- SwissProt: P12036 Human
- SwissProt: P19246 Mouse
- SwissProt: P16884 Rat
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Alternative names
- 200 kDa neurofilament protein antibody
- CMT2CC antibody
- Nefh antibody
see all
Images
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Immunohistochemistry - Free Floating - Anti-Neurofilament heavy polypeptide antibody (ab4680)Immunohistological analysis of a rat cerebellum section stained with ab4680 at a dilution 1:5,000 in red, and co-stained with rabbit pAb to GFAP at a dilution 1:5,000 in green. The blue is DAPI staining of nuclear DNA.
Following transcardial perfusion with 4% paraformaldehyde, brain was post fixed for 24 hours, cut to 45 μM, and free floating sections were stained.
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Western blot - Anti-Neurofilament heavy polypeptide antibody (ab4680)All lanes : Anti-Neurofilament heavy polypeptide antibody (ab4680) at 1/20000 dilution
Lane 1 : Rat spinal cord lysate
Lane 2 : Mouse spinal cord lysate
Lane 3 : Cow spinal cord lysate
Predicted band size: 200-220 kDa -
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Neurofilament heavy polypeptide antibody (ab4680)This image is courtesy of an Abreview submitted by Francoise Geffroy.Immunohistochemical staining of Neurofilament heavy polypeptide 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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Immunocytochemistry/ Immunofluorescence - Anti-Neurofilament heavy polypeptide antibody (ab4680)Mixed neuron/glial cultures stained with a mouse monoclonal antibody to neurofilament subunit NF-L (green) and ab4680, chicken antibody to neurofilament heavy polypeptide. This antibody binds primarily to the phosphorylated axonal forms of NF-H, in contrast to the NF-L antibody which stains both axonal and dendritic/perikaryal neurofilaments. The NF-L antibody therefore reveals a prominent cell body in green, while the surrounding axonal profiles are orange, since the are bound by both NF-L and the chicken NF-H antibody. Blue is a DNA stain.
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Immunocytochemistry/ Immunofluorescence - Anti-Neurofilament heavy polypeptide 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). -
Immunocytochemistry/ Immunofluorescence - Anti-Neurofilament heavy polypeptide antibody (ab4680)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).
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Immunohistochemistry (Frozen sections) - Anti-Neurofilament heavy polypeptide antibody (ab4680)This image is courtesy of an anonymous Abreview.ab4680 at 1/10000 dilution staining Neurofilament heavy polypeptide 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.
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Immunocytochemistry/ Immunofluorescence - Anti-Neurofilament heavy polypeptide antibody (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.201003048ab4680 staining Neurofilament heavy polypeptide (green) in murine embryoid bodies by Immunocytochemistry/ Immunofluorescence. Nuclei were visualized with DAPI.
References
This product has been referenced in:
- Brick RM et al. Neurotrophically Induced Mesenchymal Progenitor Cells Derived from Induced Pluripotent Stem Cells Enhance Neuritogenesis via Neurotrophin and Cytokine Production. Stem Cells Transl Med 7:45-58 (2018). Read more (PubMed: 29215199) »
- Hill RZ et al. The signaling lipid sphingosine 1-phosphate regulates mechanical pain. Elife 7:N/A (2018). Read more (PubMed: 29561262) »
