Antibody was affinity purified using an epitope specific to Phospho MCM2-S53 immobilized on solid support. Immunogen was a synthetic phosphorylated peptide, which represented a portion of human minichromosomal maintenance deficient 2 surrounding Serine 53 according to the numbering given in entry NP_004517.2 (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 53
2 - 8°C
Detection of human Phospho MCM2 (S53) by western blot. Samples: Whole cell lysate (5 and 15 µg for WB; 1 mg for IP, 20% of IP loaded) from asynchronous HEK293T cells. For IP/WB, MCM2 was immunoprecipitated using A300-191A, which recognizes total MCM2. The immunoprecipitate was mock treated (-) or treated (+) with phosphatase (PPase). Antibody: Affinity purified rabbit anti-phospho MCM2 (S53) antibody A300-756A (lot A300-756A-4) used at 0.1 µg/ml. To examine total MCM2, the membrane was stripped and blotted with A300-191A at 0.1 µg/ml. Detection: Chemiluminescence with exposure times of 10 seconds (A and B).
Detection of human and mouse Phospho MCM2 (S53) by immunohistochemistry. Sample: FFPE sections of human breast carcinoma (left) and mouse renal cell carcinoma (right). Antibody: Affinity purified rabbit anti-phospho MCM2 (S53) (Cat. No. A300-756A Lot4) used at a dilution of 1:200 (1µg/ml). Detection: DAB