Antibody was affinity purified using an epitope specific to Chk1 immobilized on solid support. The epitope recognized by A300-298A maps to a region between residues 250 and 300 of human Checkpoint Kinase 1 using the numbering given in SwissProt entry O14757 (GeneID 1111). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
cell cycle checkpoint kinase, Checkpoint kinase-1, Checkpoint, S. pombe, homolog of, 1, CHK1, CHK1 checkpoint homolog, Chk1-S, serine/threonine-protein kinase Chk1
between 250 and 300
2 - 8°C
Tris-citrate/phosphate buffer, pH 7 to 8 containing 0.09% Sodium Azide
Detection of human Chk1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, Jurkat, and mouse TMCK cells. Antibodies: Affinity purified rabbit anti-Chk1 antibody A300-298A (lot A300-298A-2) used for WB at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human Chk1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP; 20% of IP loaded) from HeLa cells. Antibodies: Affinity purified rabbit anti-Chk1 antibody A300-298A (lot A300-298A-2) used for IP at 6 µg/mg lysate. Chk1 was also immunoprecipitated by a previous lot (lot A300-298A-1) of this antibody. For blotting immunoprecipitated Chk1, A300-298A-2 was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.