Antibody was affinity purified using a synthetic peptide representing phosphorylation at Serine 108 and surrounding residues of MCM2 immobilized on solid support. Immunogen A300-094A was a synthetic phosphorylated peptide, which represented a portion of human Minichromosomal Maintenance Deficient 2 surrounding Serine 108 according to the numbering given in entry NP_0045117.2 (GeneID 4171). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
BM28, CCNL1, cdc19, CDCL1, cell devision cycle-like 1, cyclin-like 1, D3S3194, DFNA70, DNA replication licensing factor MCM2, minichromosome maintenance deficient 2 (mitotin), minichromosome maintenance protein 2 homolog, MITOTIN, nuclear protein BM28
surrounding serine 108
2 - 8°C
Detection of human and mouse Phospho MCM2 (S108) by western blot. Samples: Whole cell lysate (15 or 50 µg) from HEK293T cells or mouse NIH 3T3 (M) cells treated with 100 µM etoposide for 2 hours (+) or mock treated (-) cells. Antibodies: Affinity purified rabbit anti-phospho MCM2 (S108) antibody A300-094A (lot A300-094A-4) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.
Detection of human Phospho MCM2 (S108) by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP; 20% of IP loaded) from HEK293T cells either mock treated (-) or treated (+) with etoposide (100 µM, 2h). Antibodies: MCM2 was immunoprecipitated using rabbit anti-MCM2 antibody A300-191A. Following immunoprecipitation the product of the IP reaction was either mock treated (-) or treated with phosphatases (+). Affinity purified rabbit anti-phospho MCM2 (S108) antibody A300-094A (lot A300-094A-4) was used for WB at 1 µg/ml. To examine total MCM2, the blot was stripped and then blotted with rabbit anti-MCM2 antibody A300-191A. Detection: Chemiluminescence with exposure times of 10 seconds (upper and lower panels).
Detection of human and mouse Phospho MCM2 (S108) by immunohistochemistry. Sample: FFPE sections of human breast carcinoma (left) and mouse renal cell carcinoma (right). Antibody: Affinity purified rabbit anti- phospho MCM2 (S108) (Cat. No. A300-094A Lot4) used at a dilution of 1:5,000 (0.2µg/ml). Detection: DAB