Anti-alpha Internexin antibody [2E3] (ab7654)

Overview

  • Product nameAnti-alpha Internexin antibody [2E3]
    See all alpha Internexin primary antibodies
  • Description
    Mouse monoclonal [2E3] to alpha Internexin
  • Specificityspecifically recognizes a-internexin.
  • Tested applicationsSuitable for: IHC-Fr, WB, Flow Cyt, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: all Mammals
  • Immunogen

    Recombinant full length protein (Rat).

  • EpitopeC-terminal non-helical extension of the protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab7654 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.
WB 1/100. Predicted molecular weight: 66 kDa.
Flow Cyt 1/100.
ICC/IF 1/500.
IHC-P 1/5000. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionClass-IV neuronal intermediate filament that is able to self-assemble. It is involved in the morphogenesis of neurons. It may form an independent structural network without the involvement of other neurofilaments or it may cooperate with NF-L to form the filamentous backbone to which NF-M and NF-H attach to form the cross-bridges.
  • Tissue specificityFound predominantly in adult CNS.
  • Sequence similaritiesBelongs to the intermediate filament family.
  • Developmental stageExpressed in brain as early as the 16th week of gestation, and increased rapidly and reached a steady state level by the 18th week of gestation.
  • Post-translational
    modifications
    O-glycosylated.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Information by UniProt
  • Database links
  • Alternative names
    • 66 kDa neurofilament protein antibody
    • AINX_HUMAN antibody
    • Alpha Inx antibody
    • Alpha-internexin antibody
    • Alpha-Inx antibody
    • INA antibody
    • Internexin neuronal intermediate filament protein alpha antibody
    • MGC12702 antibody
    • NEF 5 antibody
    • NEF5 antibody
    • Neurofilament 5 (66kD) antibody
    • Neurofilament 5 antibody
    • Neurofilament 66 antibody
    • Neurofilament 66 tax binding protein antibody
    • Neurofilament-66 antibody
    • NF 66 antibody
    • NF-66 antibody
    • NF66 antibody
    • TXBP 1 antibody
    • TXBP1 antibody
    see all

Anti-alpha Internexin antibody [2E3] images

  • Hippocampal neurons in tissue culture with neurofilament alpha internexin (red) (Ab7654) and GFAP (green). Photo courtesy of: Dr. Gerry Shaw University of Florida
  • All lanes : Anti-alpha Internexin antibody [2E3] (ab7654)

    Lane 1 : Crude homogenate of rat facial nucleus at 3 days after experimental injury
    Lane 2 : Crude homogenate of rat facial nucleus at 3 days after experimental injury
    Lane 3 : Crude homogenate of rat facial nucleus at 5 days after experimental injury
    Lane 4 : Crude homogenate of rat facial nucleus at 5 days after experimental injury
    Lane 5 : Crude homogenate of rat facial nucleus at 7 days after experimental injury
    Lane 6 : Crude homogenate of rat facial nucleus at 7 days after experimental injury
    Lane 7 : Crude homogenate of rat facial nucleus at 14 days after experimental injury
    Lane 8 : Crude homogenate of rat facial nucleus at 14 days after experimental injury
    Lane 9 : Crude homogenate of rat facial nucleus at 28 days after experimental injury
    Lane 10 : Crude homogenate of rat facial nucleus at 28 days after experimental injury


    Predicted band size : 66 kDa
    Observed band size : 66 kDa
  • Overlay histogram showing SH-SY5Y cells stained with ab7654 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab7654, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab7654 staining alpha Internexin in human cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% hydrogen peroxide for 10 minutes at 25°C; antigen retrieval was by heat mediation in EDTA, pH 9.0 (100°C for 20 minutes). Samples were incubated with primary antibody (1/2500) for 20 minutes at 25°C.

    See Abreview

References for Anti-alpha Internexin antibody [2E3] (ab7654)

This product has been referenced in:
  • Rauch U  et al. Expression of intermediate filament proteins and neuronal markers in the human fetal gut. J Histochem Cytochem 54:39-46 (2006). Read more (PubMed: 16087706) »
  • Uchikado H  et al. Screening for neurofilament inclusion disease using alpha-internexin immunohistochemistry. Neurology 64:1658-9 (2005). Read more (PubMed: 15883343) »

See all 3 Publications for this product

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step hydrogen peroxide as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA, pH9.0, 100C, 20 min
Sample Human Tissue sections (cerebral cortex)
Specification cerebral cortex
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 01 2014

Thank you for contacting us about this issue. I am tempted to say you are detecting internexin +/- phosphorylation but the bands appear to be too far apart to fit that explanation. Do you have an image with the molecular weight markers, assuming you in...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Loading amount 50 µg
Specification Brain
Gel Running Conditions Reduced Denaturing (4-15%)
Blocking step Milk as blocking agent for 14 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Dec 06 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Apteronotus leptorhynchus Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% Triton X-100
Blocking step 3% sheep serum, 1% BSA, 1% teleostean gelatine in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Username

Abcam user community

Verified customer

Submitted Apr 12 2012


I am very pleased to hear you would like to accept our offer and test the above antivbodies in Apteronotus leptorhynchus. This code will give you 1 free primary antibody before the expiration date for each code. To redeem this offer, please submit...

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