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  1. Link

    involucrin-antibody-sy5-ab68.pdf

  1. Send me a copy of this email
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Signal Transduction Cytoskeleton / ECM Extracellular Matrix Structures Keratin
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Anti-Involucrin antibody [SY5] (ab68)

  • Datasheet
  • SDS
Reviews (9)Q&A (7)References (40)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)
  • Flow Cytometry - Anti-Involucrin antibody [SY5] (ab68)
  • Immunocytochemistry/ Immunofluorescence - Anti-Involucrin antibody [SY5] (ab68)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)

Key features and details

  • Mouse monoclonal [SY5] to Involucrin
  • Suitable for: ICC/IF, IHC-P, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-Involucrin antibody [SY5]
    See all Involucrin primary antibodies
  • Description

    Mouse monoclonal [SY5] to Involucrin
  • Host species

    Mouse
  • Tested applications

    Suitable for: ICC/IF, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to Human Involucrin. Purified involucrin from human keratinocytes.

  • Epitope

    The epitope maps between codon 421-568 of human involucrin.
  • Positive control

    • IHC-P: Human tonsil, skin tissue. Localized to upper spinous and granular layers in normal skin. Flow cyt: A431 cells. ICC/IF: MCF7 cells.
  • 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.

    Staining of formalin-fixed tissues is enhanced by digestion with trypsin or Protease XXV at 1mg/ml PBS for 5 min at 37°C (Note than enzyme digestion is better than citrate buffer based epitope unmasking)

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Monoclonal
  • Clone number

    SY5
  • Myeloma

    P3x63
  • Isotype

    IgG1
  • Light chain type

    unknown
  • Research areas

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

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)

Applications

The Abpromise guarantee

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.
IHC-P (2)
Use a concentration of 0.1 - 0.2 µg/ml.
Flow Cyt
Use 1µg for 106 cells.

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

Notes
ICC/IF
Use a concentration of 1 µg/ml.
IHC-P
Use a concentration of 0.1 - 0.2 µg/ml.
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.
  • 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 [SY5] (ab68)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)This image is courtesy of an anonymous abreview.
      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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)Chavez-Munoz, C. et al PLoS One. 2013 Dec 2;8(12):e80587. doi: 10.1371/journal.pone.0080587. eCollection 2013 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      KLC (keratinocyte-like cells) have the capacity to stratify

      Human skin was decellularized and confirmed complete cell removal by H&E staining. ASC (human adipose-derived stem cells) were seeded on the epithelial side of human dermal decellularized matrices in the presence or absence of EGF (0.005 µg/mL). Human dermal decellularized matrices seeded with keratinocytes were used as controls. All three groups were kept in the same media conditions and lifted to an air-liquid interphase at the same time. Histological analysis revealed the presence of several layers of KLC with an organization similar to that of keratinocytes (A.1and 3). Surprisingly, ASC without EGF not only did not transdifferentiate into KLC, but instead migrated inside the matrix (A.2). Immunohistochemical analysis revealed a positive staining of cytokeratin-5 (B.3), cytokeratin-10 (C.3) and Involucrin (D.3) in stratified KLC matrices comparable to that of the stratified keratinocyte matrix (A, B, C and D.1). No positive staining was found in the ASC seeded matrices (A, B, C and D.2).

    • Flow Cytometry - Anti-Involucrin antibody [SY5] (ab68)
      Flow Cytometry - Anti-Involucrin antibody [SY5] (ab68)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Involucrin antibody [SY5] (ab68)
      Immunocytochemistry/ Immunofluorescence - Anti-Involucrin antibody [SY5] (ab68)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)

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

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (40)

    Publishing research using ab68? Please let us know so that we can cite the reference in this datasheet.

    ab68 has been referenced in 40 publications.

    • De Tollenaere M  et al. Overall renewal of skin lipids with Vetiver extract for a complete anti-ageing strategy. Int J Cosmet Sci 43:165-180 (2021). PubMed: 33253416
    • Abreu CM  et al. Microscopy-guided laser ablation for the creation of complex skin models with folliculoid appendages. Bioeng Transl Med 6:e10195 (2021). PubMed: 34027085
    • Katekawa E  et al. Novel topical skin hydration agent containing Anadenanthera colubrina polysaccharide-standardized herbal preparation. J Cosmet Dermatol 19:1691-1698 (2020). PubMed: 31743554
    • Robinson DJ  et al. Epigenetic regulation of iASPP-p63 feedback loop in cutaneous squamous cell carcinoma. J Invest Dermatol N/A:N/A (2019). PubMed: 30710576
    • Egolf S  et al. LSD1 Inhibition Promotes Epithelial Differentiation through Derepression of Fate-Determining Transcription Factors. Cell Rep 28:1981-1992.e7 (2019). PubMed: 31433976
    View all Publications for this product

    Customer reviews and Q&As

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

    1-10 of 16 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-Involucrin antibody [SY5]

    Inconclusive
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (skin)
    Permeabilization
    No
    Specification
    skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
    Fixative
    Acetone
    Read More

    Mrs. Laurène ROMANO

    Verified customer

    Submitted May 27 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Involucrin antibody [SY5]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (NHEKs (keratinocytes))
    Permeabilization
    No
    Specification
    NHEKs (keratinocytes)
    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 Nov 07 2019

    Immunohistochemistry (Frozen sections) abreview for Anti-Involucrin antibody [SY5]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 14 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-Involucrin antibody [SY5]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 20 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Involucrin antibody [SY5]

    Poor
    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

    Question

    mouse tonsil

    1) Why are bands sen at 120 kDa and 170 kDa with human samples?
    2) Why is the antibody not good forouse? Data?
    3) send database links to human and mouse protein

    Read More

    Abcam community

    Verified customer

    Asked on Aug 10 2012

    Answer

    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.

    The antibody was tested against mouse but showed no reactivity."

    I hope this information is nevertheless helpful to you.
    If you are not satisfied with the antibody, I'd be happy to replace, credit or refund it - if it was used in a tested species or application.Please do let me know.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 10 2012

    Question

    precisions about the epitope, what is the aa sequence?
    possibility to have the full lenght protein?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 10 2012

    Answer

    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 for this antibody.
    We are not able to supply the protein but should that ever be possible we will get in touch."

    I am sorry I could not be more helpful on this occasion. Please do not hesitate to contact me again if you have further questions.

    Read More

    Abcam Scientific Support

    Answered on Aug 10 2012

    Question

    mouse tonsil

    1) Why are bands sen at 120 kDa and 170 kDa with human samples?
    2) Why is the antibody not good forouse? Data?
    3) send database links to human and mouse protein

    Read More

    Abcam community

    Verified customer

    Asked on Jul 07 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 07 2012

    Question

    mouse tonsil

    1) Why are bands seen at 120 kDa and 170 kDa with human samples?
    2) Why is the antibody not good for mouse? Data?
    3) send database links to human and mouse protein

    Read More

    Abcam community

    Verified customer

    Asked on Jul 03 2012

    Answer

    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/P07476
    mouse: http://www.uniprot.org/uniprot/P48997

    I hope this information is of some help to you.
    In the meantime, please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Jul 03 2012

    Question

    Hello I am trying to gather MSDs’s all chemicals we keep for health and safety purposes, and One of them is : Involucrin mouse monoclonal cat no: ab68 Could you provise me with this document cause it is not accessible on the website. Thank you

    Read More

    Abcam community

    Verified customer

    Asked on Feb 13 2012

    Answer

    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.

    Read More

    Abcam Scientific Support

    Answered on Feb 13 2012

    1-10 of 16 Abreviews or Q&A

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