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Recombinant fragment, corresponding to amino acids 60-383 of Human KAP1.
Our Abpromise guarantee covers the use of ab22553 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent dilution.|
|IHC-P||Use a concentration of 5 µg/ml.|
|ICC/IF||Use a concentration of 2 - 4 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 88.5 kDa).|
|EMSA||Use at an assay dependent dilution.|
|IP||Use at an assay dependent concentration.|
Immunofluorescence analysis of A549 cells, staining KAP1 (green) with ab22553.
Cells were fixed in 4% paraformaldehyde followed by permeabilization in 0.1% Triton X-100/PBS. Cells were incubated in primary antibody and a FITC-conjugated goat anti-mouse secondary antibody was used to detect staining. Nuclei were stained with DAPI (blue).
IHC image of KAP1 staining in Human normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab22553, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer 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.