The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml.
1/15 - 1/50. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.
Use at an assay dependent concentration.
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.
Anti-pan Cytokeratin antibody [5D3 + LP34] images
Immunocytochemistry/ Immunofluorescence - Anti-pan Cytokeratin antibody [5D3 + LP34] (ab17153)This image is courtesy of an Abreview submitted by Gabriel Ortega
ab17153 staining pan Cytokeratin (green) in Human cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Tween 20 and blocked with 10% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. ab150113, a goat anti-mouse Alexa Fluor®488 (IgG polyclonal; 1/100) was used as the secondary antibody.
ICC/IF image of ab17153 stained HeLa 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 (ab17153, 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.
Western blot - pan Cytokeratin antibody [5D3 + LP34] (ab17153)This image is courtesy of an anonymous Abreview
Anti-pan Cytokeratin antibody [5D3 + LP34] (ab17153) at 1/500 dilution ((in 5% Marvel TBS-T). Incubation for 1 hour at 22°C) + Whole cell lysate of human HT1080 cells at 25 µg
Secondary An HRP-conjugated Sheep anti-mouse IgG monoclonal at 1/5000 dilution developed using the ECL technique
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan Cytokeratin antibody [5D3 + LP34] (ab17153)This image is courtesy of an Abreview submitted by Carl Hobbs
ab17153 staining human skin and lung sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab17153 at 1/250 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-mouse polyclonal antibody at 1/200 was used as the secondary antibody.