The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/1000 - 1/2500. Detects a band of approximately 117 kDa (predicted molecular weight: 100 kDa).
Acts as component of the MCM2-7 complex (MCM complex) which is the putative replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. The active ATPase sites in the MCM2-7 ring are formed through the interaction surfaces of two neighboring subunits such that a critical structure of a conserved arginine finger motif is provided in trans relative to the ATP-binding site of the Walker A box of the adjacent subunit. The six ATPase active sites, however, are likely to contribute differentially to the complex helicase activity. Required for the entry in S phase and for cell division.
Belongs to the MCM family. Contains 1 MCM domain.
Phosphorylated on Ser-108 by ATR in proliferating cells. Ser-108 proliferation is increased by genotoxic agents. Ser-40 is mediated by the CDC7-DBF4 and CDC7-DBF4B complexes, while Ser-53 phosphorylation is only mediated by the CDC7-DBF4 complex. Phosphorylation by the CDC7-DBF4 complex during G1/S phase is required for the initiation of DNA replication.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma (left) and mouse renal cell carcinoma (right) tissues labelling MCM2 (phospho S53) with ab70367 at 1/200 (1µg/ml). Detection: DAB.
Western blot - MCM2 (phospho S53) antibody (ab70367)
All lanes : Anti-MCM2 (phospho S53) antibody (ab70367) at 0.1 µg/ml
Lane 1 : Whole cell lysate from asynchronous 293T cells Lane 2 : Whole cell lysate from asynchronous 293T cells treated with a phosphatase.
Lysates/proteins at 50 µg per lane.
Predicted band size : 100 kDa Observed band size : 117 kDa (why is the actual band size different from the predicted?) The bottom panel shows total MCM2 staining; the blot was stripped and then blotted with rabbit anti-total MCM2 antibody at 0.1 µg/ml. Detection: Chemiluminescence with exposure times of 3 minutes (upper panel) and 10 seconds (lower panel).