The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
1/10000. Detects a band of approximately 81 kDa (predicted molecular weight: 81 kDa).
Use a concentration of 5 µg/ml.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration.
FunctionActs 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 S-phase checkpoint activation upon UV-induced damage.
Sequence similaritiesBelongs to the MCM family. Contains 1 MCM domain.
Post-translational modificationsPhosphorylated upon DNA damage, probably by ATM or ATR.
ab52489 staining MCM7 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.
MCM7 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to MCM7 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52489.
Overlay histogram showing HeLa cells stained with ab52489 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52489, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Western blot - MCM7 antibody [EP1974Y] (ab52489)
Anti-MCM7 antibody [EP1974Y] (ab52489) at 1/10000 dilution + Hela cell lysate at 10 µg
Secondary Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size : 81 kDa Observed band size : 81 kDa
Immunohistochemical analysis of paraffin-embedded human lung tissue using ab52489 at a 1/100 dilution.
References for Anti-MCM7 antibody [EP1974Y] (ab52489)
This product has been referenced in:
Xu X et al. TIMELESS Suppresses the Accumulation of Aberrant CDC45·MCM2-7·GINS Replicative Helicase Complexes on Human Chromatin. J Biol Chem291:22544-22558 (2016).
Read more (PubMed: 27587400) »
Quimbaya M et al. Deregulation of the Replisome Factor MCMBP Prompts Oncogenesis in Colorectal Carcinomas through Chromosomal Instability. Neoplasia16:694-709 (2014).
Read more (PubMed: 25246271) »