Key features and details
- Mouse monoclonal [SY5] to Involucrin
- Suitable for: ICC/IF, IHC-P, Flow Cyt
- Reacts with: Dog, Human, Pig, Gorilla, Owl monkey
- Isotype: IgG1
参阅全部 Involucrin 一抗
描述小鼠单克隆抗体[SY5] to Involucrin
经测试应用适用于: ICC/IF, IHC-P, Flow Cytmore details
种属反应性与反应: Dog, Human, Pig, Gorilla, Owl monkey
Pure involucrin from human keratinocytes.
表位The epitope maps between codon 421-568 of human involucrin.
- IHC-P: Human tonsil, skin tissue. Localized to upper spinous and granular layers in normal skin. Flow cyt: A431 cells. ICC/IF: MCF7 cells.
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)
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
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存放说明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.
存储溶液Preservative: 0.02% Sodium azide
Concentration information loading...
纯度Immunogen affinity purified
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.
|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.
功能Part of the insoluble cornified cell envelope (CE) of stratified squamous epithelia.
组织特异性Keratinocytes of epidermis and other stratified squamous epithelia.
序列相似性Belongs to the involucrin family.
翻译后修饰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.
细胞定位Cytoplasm. Constituent of the scaffolding of the cornified envelope.
- Information by UniProt
- INVO_HUMAN antibody
- Involucrin antibody
- IVL antibody
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).
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).
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.
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.
Immunohistochemical analysis of Formalin fixed paraffin embedded human tonsil section labelling Involucrin with ab68 at dilution of 0.1 ug/mL.
ab68 被引用在 32 文献中.
- 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
- Okazaki S et al. Glutaminolysis-related genes determine sensitivity to xCT-targeted therapy in head and neck squamous cell carcinoma. Cancer Sci 110:3453-3463 (2019). PubMed: 31444923
- 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). PubMed: 30093649
- Rosato B et al. Role of FGFR2b expression and signaling in keratinocyte differentiation: sequential involvement of PKCd and PKCa. Cell Death Dis 9:565 (2018). PubMed: 29752438