Overview

  • Product name
    Anti-Involucrin antibody [SY5]
    See all Involucrin primary antibodies
  • Description
    Mouse monoclonal [SY5] to Involucrin
  • Tested applications
    Suitable for: ICC/IF, WB, IP, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Dog, Human, Pig, Gorilla, Owl monkey
    Does not react with: Mouse
  • Immunogen

    Pure involucrin from human keratinocytes.

  • Epitope
    The epitope maps between codon 421-568 of human involucrin.
  • Positive control
    • MCF-7 cells. Localized to upper spinous and granular layers in Normal skin.
  • General notes

    Involucrin is the differentiation marker of human keratinocytes.
    ab81216 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab68 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 a concentration of 1 µg/ml.
WB Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 0.1 - 0.2 µg/ml.
IHC-Fr 1/100.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

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

  • Immunohistochemical analysis of Formalin fixed paraffin embedded human tonsil section labelling Involucrin with ab68 at dilution of 0.1 ug/mL. 

  • ab68 at a 1/1000 dilution staining Involucrin in human normal skin tissue by Immunohistochemistry (formalin fixed, paraffin embedded sections), incubated for 2 hours at 21°C. Heat mediated antigen retrieval step performed using citrate buffer pH 6.0. Blocked with 10% serum for 30 minutes at 21°C. Secondary used at 1/100 polyclonal Donkey anti-mouse IgG conjugated to Alexa Fluor 594 (red).

    See Abreview

  • Immunohistochemical analysis of frozen Human artifical skin tissue labeling Involucrin with ab68 at 1/200 dilution.

    See Abreview

  • ICC/IF image of ab68 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab68, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing A431 cells stained with ab68 (red line). The cells were fixed with 80% methanol (5 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 (ab68, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H&L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Dziunycz PJ  et al. CYFIP1 is directly controlled by NOTCH1 and down-regulated in cutaneous squamous cell carcinoma. PLoS One 12:e0173000 (2017). WB ; Human . Read more (PubMed: 28410392) »
  • Kim BR  et al. SOX2 and PI3K Cooperate to Induce and Stabilize a Squamous-Committed Stem Cell Injury State during Lung Squamous Cell Carcinoma Pathogenesis. PLoS Biol 14:e1002581 (2016). ICC/IF ; Human . Read more (PubMed: 27880766) »

See all 23 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 21°C
Fixative
Acetone
Username

Abcam user community

Verified customer

Submitted Jul 14 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (artificial skin)
Permeabilization
Yes - Triton X-100
Specification
artificial skin
Blocking step
Blocking One as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 20 2015

Thank you for your patience.
I apologize for the delay, but I only heard back from the supplying lab today with the following information:

"Wehave unfortunately no data to explain the differeing bands seen against human tissue.
Read More

I am very sorry for the delayed response.

Despite numerous reminders the source of the antibody only replied to me today with the following information:

"We have no other information other than the codon sequence on the epitope f...

Read More

Thank you for contacting us.

I apologize for the wait, but I have not yet heard back from the lab. I have sent another email to follow up with them.

Thank you for your patience and understanding.

Thank you for contacting us.

I have contacted the lab for the questions you had, and will let you know their answer.

The database links for human and mouse proteins are as follows:

human: http://www.uniprot.org/uniprot/P07...

Read More
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
Username

Mr. Carl Hobbs

Verified customer

Submitted Mar 19 2012

Thank you for contacting us.

I have attached the MSDS for ab68.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Skin)
Specification
Skin
Fixative
Acetone
Permeabilization
No
Blocking step
Casein as blocking agent for 10 minute(s) · Concentration: 0.25% · Temperature: 20°C
Username

Dr. Vaughan Feisst

Verified customer

Submitted Oct 23 2010

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 (Normal Skin)
Specification
Normal Skin
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrat, pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Feb 19 2010

1-10 of 14 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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