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 - 1/5000. Detects a band of approximately 117 kDa (predicted molecular weight: 100 kDa).
Use 0.1µg for 106 cells.
0.1 μg per 1 X 106 cells in a 150 µl volume.
ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
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 colon carcinoma mock phosphatase treated section (left) and calf intestinal phosphatase-treated section (right) tissues labelling MCM2 (phospho S40 + S41) with ab70371 at 1/200 (1µg/ml). Detection: DAB.
Western blot - MCM2 (phospho S40 + S41) antibody (ab70371)
All lanes : Anti-MCM2 (phospho S40 + S41) antibody (ab70371) 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?) Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.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 30 seconds for upper and lower panel.
Erbayraktar Z et al. Cell division cycle 7-kinase inhibitor PHA-767491 hydrochloride suppresses glioblastoma growth and invasiveness. Cancer Cell Int16:88 (2016).
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