Antibodies were affinity purified using epitopes specific to PLK1 immobilized on solid support. The epitopes recognized by A300-251A maps to a region between residues 325 and 375 of human Polo-like kinase using the numbering given in SwissProt entry P53350 (GeneID 5347). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
cell cycle regulated protein kinase, PLK, PLK-1, polo (Drosophia)-like kinase, Polo-like kinase 1, serine/threonine-protein kinase 13, serine/threonine-protein kinase PLK1, STPK13
between 325 and 375
2 - 8°C
Detection of human PLK1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-PLK1 antibody A300-251A (lot A300-251A-1) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
Detection of human PLK1 by immunocytochemistry. Sample: Formaldehyde-fixed asynchronous HeLa cells. Antibody: Affinity purified rabbit anti- PLK1 (Cat. No.A300-251A Lot1) used at a dilution of 1:1,000 (1µg/ml). Detection: Green fluorescent goat anti-rabbit IgG highly cross-adsorbed Antibody FITC conjugated (Cat. No. A120-501F) used at a dilution of 1:100.
Detection of human PLK1 by western blot and immunoprecipitation. Samples: Whole cell lysate (50 µg for A; 600 µg for B) from HeLa cells. Antibodies: A. Affinity purified rabbit anti-PLK1 antibody A300-251A used at the indicated concentrations for WB. B. Affinity purified rabbit anti-PLK1 antibodies A300-250A, A300-251A, and BL978 used at 2 µg/600 µg lysate for IP. immunoprecipitatesd PLK1 was detected using BL978 at 0.5 µg/ml. Detection: Chemiluminescence with exposure times of less than 5 minutes.