Overview

  • Product name
  • Description
    Rabbit polyclonal to Involucrin
  • Host species
    Rabbit
  • Specificity
    ab53112 detects endogenous levels of total Involucrin protein.
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-P, ELISAmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic peptide derived from human Involucrin.

  • Positive control
    • Human breast carcinoma tissue and extracts from HuvEc cells.

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

Applications

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.

Application Abreviews Notes
ICC/IF Use at an assay dependent concentration. PubMed: 21067603
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.
ELISA 1/20000.

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.
  • Information by UniProt
  • Database links
  • Alternative names
    • INVO_HUMAN antibody
    • Involucrin antibody
    • IVL antibody

Images

  • 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

  • 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

  • Immunofluorescence analysis of Human skin xenografts carrying either control GFP (left) or SHH retrovirus (right), staining Involucrin (red) with 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.

References

This product has been referenced in:
  • Loh EYX  et al. Development of a bacterial cellulose-based hydrogel cell carrier containing keratinocytes and fibroblasts for full-thickness wound healing. Sci Rep 8:2875 (2018). IHC . Read more (PubMed: 29440678) »
  • Chen J  et al. Epidermal CFTR Suppresses MAPK/NF-?B to Promote Cutaneous Wound Healing. Cell Physiol Biochem 39:2262-2274 (2016). Read more (PubMed: 27832634) »
See all 6 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

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

Abcam user community

Verified customer

Submitted Aug 01 2017

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

Mr. Julien Coutier

Verified customer

Submitted Jun 20 2017

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

Abcam user community

Verified customer

Submitted Dec 15 2015

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

Abcam user community

Verified customer

Submitted Aug 12 2014

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 guarantees this product to work in the species/application used in this Abreview.
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

Mr. Carl Hobbs

Verified customer

Submitted Mar 19 2012

Abcam has not validated the combination of species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Feb 01 2012

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

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