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  1. Link

    involucrin-antibody-ab53112.pdf

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Signal Transduction Cytoskeleton / ECM Extracellular Matrix Structures Keratin
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Anti-Involucrin antibody (ab53112)

  • Datasheet
  • SDS
Reviews (7)Q&A (2)References (16)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)
  • Western blot - Anti-Involucrin antibody (ab53112)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)

Key features and details

  • Rabbit polyclonal to Involucrin
  • Suitable for: WB, IHC-P
  • Reacts with: Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Involucrin antibody
    See all Involucrin primary antibodies
  • Description

    Rabbit polyclonal to Involucrin
  • Host species

    Rabbit
  • Specificity

    ab53112 detects endogenous levels of total Involucrin protein.
  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    A synthetic peptide derived from human Involucrin.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS

    Without Mg+2 and Ca+2
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    ab53112 was affinity purified from rabbit antiserum by affinity chromatography using epitope-specific immunogen.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Keratin
    • Cancer
    • Tumor biomarkers
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab53112 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
IHC-P
Human
WB
Human
Application Abreviews Notes
WB
1/300 - 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
IHC-P (5)
Use at an assay dependent concentration.
Notes
WB
1/300 - 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
IHC-P
Use at an assay dependent concentration.

Target

  • Function

    Part of the insoluble cornified cell envelope (CE) of stratified squamous epithelia.
  • Tissue specificity

    Keratinocytes of epidermis and other stratified squamous epithelia.
  • Sequence similarities

    Belongs to the involucrin family.
  • Post-translational
    modifications

    Substrate of transglutaminase. Some glutamines and lysines are cross-linked to other involucrin molecules, to other proteins such as keratin, desmoplakin, periplakin and envoplakin, and to lipids like omega-hydroxyceramide.
  • Cellular localization

    Cytoplasm. Constituent of the scaffolding of the cornified envelope.
  • Target information above from: UniProt accession P07476 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3713 Human
    • Omim: 147360 Human
    • SwissProt: P07476 Human
    • Unigene: 516439 Human
    • Alternative names

      • INVO_HUMAN antibody
      • Involucrin antibody
      • IVL antibody

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)This image is courtesy of an Abreview submitted by Carl Hobbs

      ab53112 staining human skin sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab53112 at 1/150 in TBS/BSA/azide for 16h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

      See Abreview

    • Western blot - Anti-Involucrin antibody (ab53112)
      Western blot - Anti-Involucrin antibody (ab53112)
      All lanes : Anti-Involucrin antibody (ab53112) at 1/300 dilution

      Lane 1 : HuvEc cell extract
      Lane 2 : HuvEc cell extract with immunising peptide

      Predicted band size: 68 kDa
      Observed band size: 68 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody (ab53112)
      ab53112 at 1/50 dilution staining Involucrin in human breast carcinoma by Immunohistochemistry, Paraffin embedded tissue, in the absence and presence of the immunising peptide.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (16)

    Publishing research using ab53112? Please let us know so that we can cite the reference in this datasheet.

    ab53112 has been referenced in 16 publications.

    • Arasteh S  et al. Efficient Wound Healing Using a Synthetic Nanofibrous Bilayer Skin Substitute in Murine Model. J Surg Res 245:31-44 (2020). PubMed: 31400575
    • Fortunel NO  et al. Iterative Three-Dimensional Epidermis Bioengineering and Xenografting to Assess Long-Term Regenerative Potential in Human Keratinocyte Precursor Cells. Methods Mol Biol N/A:N/A (2019). PubMed: 31309517
    • Meunier M  et al. From stem cells protection to skin microbiota balance: Orobanche rapum extract, a new natural strategy. J Cosmet Dermatol 18:1140-1154 (2019). PubMed: 30485658
    • Ya C  et al. Substrate softness promotes terminal differentiation of human keratinocytes without altering their ability to proliferate back into a rigid environment. Arch Dermatol Res N/A:N/A (2019). PubMed: 31392392
    • Horvath S  et al. Rapamycin retards epigenetic ageing of keratinocytes independently of its effects on replicative senescence, proliferation and differentiation. Aging (Albany NY) 11:3238-3249 (2019). PubMed: 31136303
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-9 of 9 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Reconstruted Human Epidermis)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6
    Permeabilization
    No
    Specification
    Reconstruted Human Epidermis
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 17 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citraconic anhydride
    Permeabilization
    No
    Specification
    skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 01 2017

    Immunohistochemistry (Frozen sections) abreview for Anti-Involucrin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (foreskin skin section)
    Permeabilization
    No
    Specification
    foreskin skin section
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
    Fixative
    none
    Read More

    Mr. Julien Coutier

    Verified customer

    Submitted Jun 20 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ph6
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 15 2015

    Immunohistochemistry (Frozen sections) abreview for Anti-Involucrin antibody

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Skin)
    Permeabilization
    Yes - 0,3 % Triton X
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Fixative
    Perfusion fixed with paraformaldehyde and picric acid
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 12 2014

    Question

    We see no signal from capillary westerns of primary keratinocyte lysates or purified proteinat a 1/100 dilution. Please recommend an antibody that is capable of detecting involucrinfragment ab73744.
    Please recommend a membrane fractionation protocol, and controls for ab15098, 15148, and ab64482.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 26 2012

    Answer

    Thank you for calling about the issue you are having with the involucrin antibody ab53112. The amino acid sequence of the recombinant fragment that you are using as a positive control, ab73744, overlaps the sequence of the immunogen at 11 of 14 amino acids, so the antibody should be capable of detecting that fragment, assuming that at least some of the epitopes it recognizes are present on the fragment.

    I suspect the antibody is defective, if you are confident that the capillary western protocol and other materials are known to be effective. Are you confident that the secondary antibody and detection reagents are not at fault?

    For membrane isolation protocol, I recommend the protocol at the following link:

    https://www.abcam.com/index.html?pageconfig=resource&rid=11473

    For controls for the antibodies you listed, I recommend the following:

    ab15098 - Claudin - HeLa cell lysate ab29545
    ab15148 - E-Cadherin - MCF7 cell lysate ab3871
    ab64482 - Occludin - Use a human kidney lysate such as ab30203.

    I look forward to your reply. I will be happy to suggest a replacement for ab53112, if you are confident that there is no other explanation for the lack of signal for involucrin.

    Read More

    Abcam Scientific Support

    Answered on Sep 26 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Skin)
    Specification
    Skin
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
    Read More

    Mr. Carl Hobbs

    Verified customer

    Submitted Mar 19 2012

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: H-3300 Vector lab
    Permeabilization
    No
    Specification
    skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 01 2012

    Question

    Dear Madam/Sir, regarding your inquiries these are my answers, 1) I tested the antibody on human cell lines oesophageal carcinoma , (KYSE450&OE21). 2)From Cork Cancer Research Center, CORK, IRELAND. 3) Please look to the attached file for my W.B (i expect to recognize the involucrin band in just first four samples. 4) Yes i used milk as antibody diluent but only 1% conc and incubate over night. Thanks for help.

    Read More

    Abcam community

    Verified customer

    Asked on Sep 13 2010

    Answer

    Thank you for your reply with further information. The image suggests the higher band may be the correct band as it is only present in the first four samples as expected. You previously describe a higher band at 120 kDa and I assuming this is that band. We have another involucrin antibody ab17105, which detects a specific band at 170 kDa. Furthermore I have reviewed the literature and a a 120-170 kDa protein is reported in some cell lines. Therefore this higher band is likely to be the target protein. Here are some references that may be of use. I hope this information helps.

    Read More

    Abcam Scientific Support

    Answered on Sep 13 2010

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