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Synthetic peptide conjugated to KLH derived from within residues 450 to the C-terminus of Human Cytokeratin 8.
Our Abpromise guarantee covers the use of ab107115 in the following tested applications.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).|
ICC/IF image of ab107115 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab107115, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96947, goat anti-chicken IgY DyLight 488 (H+L) used at a 1/250 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
Secondary antibody - goat anti-chicken IgY HRP
IHC image of Cytokeratin 8 staining in Human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab107115, 1µg/ml, for 15 mins at room temperature. A Goat anti-Chicken biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.