Anti-Involucrin antibody [SY5] (ab68)
- Datasheet
- References (27)
- Protocols
Overview
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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, 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.
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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.
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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
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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
Associated products
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Compatible Secondaries
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Isotype control
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
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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
modificationsSubstrate 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
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Database links
- Entrez Gene: 3713 Human
- Entrez Gene: 407242 Pig
- Omim: 147360 Human
- SwissProt: P07476 Human
- SwissProt: P18175 Pig
- Unigene: 516439 Human
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Alternative names
- INVO_HUMAN antibody
- Involucrin antibody
- IVL antibody
Images
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-3280-anti-involucrin-antibody-sy5-immunohistochemistry.jpg)
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). -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-318514-anti-involucrin-antibody-sy5-immunohistochemistry.jpg)
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).
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![Flow Cytometry - Anti-Involucrin antibody [SY5] (ab68)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-200111-anti-involucrin-antibody-sy5-flow-cytometry.jpg)
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)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-3281-anti-involucrin-antibody-sy5-immunofluorescence.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Involucrin antibody [SY5] (ab68)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-262135-anti-involucrin-antibody-sy5-immunohistochemistry.jpg)
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.
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![Immunohistochemistry (Frozen sections) - Anti-Involucrin antibody [SY5] (ab68)](https://www.abcam.com/ps/products/0/ab68/Images/ab68-260888-anti-involucrin-antibody-sy5-immunohistochemistry.jpg)
Immunohistochemistry (Frozen sections) - Anti-Involucrin antibody [SY5] (ab68)Image courtesy of anonymous abreviewerImmunohistochemical analysis of frozen Human artifical skin tissue labeling Involucrin with ab68 at 1/200 dilution.
Protocols
References
This product has been referenced in:
- Robinson DJ et al. Epigenetic regulation of iASPP-p63 feedback loop in cutaneous squamous cell carcinoma. J Invest Dermatol N/A:N/A (2019). Read more (PubMed: 30710576) »
- Kim YJ et al. Topical administration of EGF suppresses immune response and protects skin barrier in DNCB-induced atopic dermatitis in NC/Nga mice. Sci Rep 8:11895 (2018). Read more (PubMed: 30093649) »
