Overview

  • Product name
    Anti-Collagen XVII antibody [EPR18614]
    See all Collagen XVII primary antibodies
  • Description
    Rabbit monoclonal [EPR18614] to Collagen XVII
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Collagen XVII aa 1300-1400. The exact sequence is proprietary.
    Database link: Q9UMD9

  • Positive control
    • WB: A431 whole cell lysate, human, mouse and rat skin lysates. IHC-P: Human esophagus, mouse skin and rat skin tissues. ICC/IF: HaCaT and A431 cells. IP: A431 whole cell lysate.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR18614
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab184996 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
WB 1/1000. Detects a band of approximately 180, 130 kDa (predicted molecular weight: 150 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/50.

Target

  • Relevance
    Unlike most collagens, collagen XVII is a transmembrane protein. Collagen XVII is a structural component of hemidesmosomes, multiprotein complexes at the dermal epidermal basement membrane zone that mediate adhesion of keratinocytes to the underlying membrane. Mutations in the gene coding for collagen XVII are associated with both generalized atrophic benign and junctional epidermolysis bullosa. Two homotrimeric forms of type XVII collagen exist. The full length form is the transmembrane protein. A soluble form, referred to as either ectodomain or LAD 1, is generated by proteolytic processing of the full length form. Two transcript variants, one resulting from alternative splicing in the 3' UTR, have been identified for this gene.
  • Cellular localization
    Cell junction, hemidesmosome. Membrane; Single-pass type II membrane protein. Note=Localized along the plasma membrane of the hemidesmosome. 120 kDa linear IgA disease antigen and 97 kDa linear IgA disease antigen: Secreted, extracellular space, extracellular matrix, basement membrane.
  • Database links
  • Alternative names
    • 180 kDa bullous pemphigoid antigen 2 antibody
    • Alpha 1 type XVII collagen antibody
    • BA16H23.2 antibody
    • BP 180 antibody
    • BP180 antibody
    • BPA 2 antibody
    • BPAG 2 antibody
    • BPAG2 antibody
    • Bullous pemphigoid antigen 2 antibody
    • COL17A1 antibody
    • Collagen 17 antibody
    • Collagen alpha 1 XVII chain antibody
    • Collagen alpha 1(XVII) chain antibody
    • Collagen alpha1 XVII chain antibody
    • Collagen type XVII alpha 1 antibody
    • Collagen XVII alpha 1 polypeptide antibody
    • CollagenXVII antibody
    • Epidermolysis bullosa junctional localisata variant antibody
    • FLJ60881 antibody
    • KIAA0204 antibody
    • LAD 1 antibody
    • LAD1 antibody
    see all

Images

  • All lanes : Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/5000 dilution

    Lane 1 : Human skin lysate
    Lane 2 : Human fetal liver lysate
    Lane 3 : Human fetal lung lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 150 kDa
    Observed band size: 130,180 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).

  • Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/5000 dilution + A431 (Human epidermoid carcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 150 kDa
    Observed band size: 130,180 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).

  • Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/2000 dilution + Mouse skin lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 150 kDa
    Observed band size: 130,180 kDa why is the actual band size different from the predicted?


    Exposure time: 2 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).

  • Anti-Collagen XVII antibody [EPR18614] (ab184996) at 1/1000 dilution + Rat skin lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 150 kDa
    Observed band size: 130,180 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15161638).

  • Immunohistochemical analysis of paraffin-embedded Human esophagus tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on squamous epithelial cells of Human esophagus is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

     

    J Biol Chem. 1991. 266, 24064-24069.

  • Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative on Human pancreas. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on squamous epithelial cells of mouse skin is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of  paraffin-embedded rat skin tissue labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on epithelial cells of rat skin is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HaCaT (Human keratinocyte cells) cells labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on HaCaT cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab184996 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2:. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labeling Collagen XVII with ab184996 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining on A431 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab184996 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Collagen XVII was immunoprecipitated from 1mg of A431 (Human epidermoid carcinoma) whole cell lysate with ab184996 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab184996 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: A431 whole cell lysate 10ug (Input).

    Lane 2: ab184996 IP in A431 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184996 in A431 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

References

This product has been referenced in:
  • Tie D  et al. Bullous Pemphigoid IgG Induces Cell Dysfunction and Enhances the Motility of Epidermal Keratinocytes via Rac1/Proteasome Activation. Front Immunol 10:200 (2019). Read more (PubMed: 30809225) »
  • Yasukochi A  et al. Regulation of collagen type XVII expression by miR203a-3p in oral squamous cell carcinoma cells. J Biochem N/A:N/A (2019). Read more (PubMed: 30918974) »
See all 4 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Human skin)
Permeabilization
Yes - 0.5% Triton TBS Ca/Azide
Specification
Human skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Mr. Tyler Crowe

Verified customer

Submitted Sep 24 2018

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (keratinocyte,adult)
Total protein in input
100 µg
Immuno-precipitation step
Protein A/G
Specification
keratinocyte,adult

Tie Duerna

Verified customer

Submitted Jun 02 2016

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