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Immunogen was a synthetic peptide, which represented a portion of human tripartite motif-containing 28 (KAP1) encoded within exon 14 (LocusLink ID 10155).
Our Abpromise guarantee covers the use of ab10484 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/10000. Detects a band of approximately 110 kDa (predicted molecular weight: 100 kDa).|
|IP||Use at 2-10 µg/mg of lysate.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||Use a concentration of 2 µg/ml.|
Cells prepared using NETN lysis buffer. Chemiluminescence detection.
Immunocytochemistry/ Immunofluorescence analysis of U2OS cells labeling KAP1 with ab10484 at 1 μg/ml. Cells were fixed with paraformaldehyde and permeabilized with 0.1% Tween. The cells were blocked with 2% BSA for 45 minutes at 25°C, followed by incubation with ab10484 at 1 μg/ml in PBS for 50 minutes at 25°C. An undiluted monoclonal Goat Anti-rabbbit IgG Alexa Fluor® 488 was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma labeling KAP1 with ab10484 at 1/5000 dilution (0.2 μg/ml). DAB detection, Hematoxylin counterstain.
Detection of Human KAP1 by Western Blot and Immunoprecipitation. Samples: A. Whole cell lysate (50 ug) from normal 293T cells (E) or 293T cells transiently transfected with a human KAP1 construct (T). B. Whole cell lysate (0.5 mg) from normal 293T cells. Antibodies: ab10484 used at 1 ug/ml for WB (A) or 2 ug/0.5 mg lysate for IP (B). Immunoprecipitated KAP1 was detected using a KAP1 antibody from another source. Detection: Chemiluminescence with a 10 minute (A) or 1 minute (B) exposure.
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