The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 21285513
Use at an assay dependent concentration.
Use a concentration of 1 - 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use 0.1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
RelevanceCytokeratins, a group comprising at least 29 different proteins, are characteristic of epithelial and trichocytic cells. Cytokeratins 1, 4, 5, 6, and 8 are members of the type II neutral to basic subfamily. Monoclonal anti cytokeratins are specific markers of epithelial cell differentiation and have been widely used as tools in tumor identification and classification. Monoclonal Anti Pan Cytokeratin (mixture) is a broadly reactive reagent, which recognizes epitopes present in most human epithelial tissues. It facilitates typing of normal, metaplastic and neoplastic cells. Synergy between the various components results in staining amplification. This enables identification of cells, which would otherwise be stained only marginally. The mixture may aid in the discrimination of carcinomas and nonepithelial tumors such as sarcomas, lymphomas and neural tumors. It is also useful in detecting micrometastases in lymph nodes, bone marrow and other tissues and for determining the origin of poorly differentiated tumors.
There are two types of cytokeratins the acidic type I cytokeratins and the basic or neutral type II cytokeratins. Cytokeratins are usually found in pairs comprising a type I cytokeratin and a type II cytokeratin. Usually the type II cytokeratins are 8kD larger than their type I counterparts.
Overlay histogram showing A431 cells stained with ab80825 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab80825, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/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 fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - pan Cytokeratin antibody [C-11] (ab80825)
Immunohistochemistry analysis of formalin-fixed, paraffin-embedded Human colon carcinomatissue using 1/50 ab80825, in conjunction with a peroxidase-conjugated secondary antibody and an AEC chromogen. Note cytoplasmic staining of tumor cells.
References for Anti-pan Cytokeratin antibody [C-11] (ab80825)
This product has been referenced in:
Herrero R et al. The biological activity of FasL in human and mouse lungs is determined by the structure of its stalk region. J Clin Invest : (2011).
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Bártek J et al. A series of 14 new monoclonal antibodies to keratins: characterization and value in diagnostic histopathology. J Pathol164:215-24 (1991).
Read more (PubMed: 1716305) »