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Anti-MCM7 Antibody EP1974Y
Also for MCM7 (NM_005916)
|A synthetic peptide corresponding to residues near the C-terminus of human MCM7 was used as an immunogen.|
|Mouse, Rat, Human
||Lot dependent; please refer to CoA along with shipment
|WB, IHC, IF, IP, FC
||ICC/IF: Use a concentration of 1 ug/ml; WB: 1:10000; IP: Use a concentration of 5 ug/ml; ICC: 1:100 - 1:250; FC: 1:100; IHC-P: Use at an assay dependent concentration
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant
|Homo sapiens minichromosome maintenance complex component 7 (MCM7), transcript variant 1|
|CDC47; MCM2; P1.1-MCM3; P1CDC47; P85MCM; PNAS146; PPP1R104|
Entrez Gene 4176 Human
Entrez Gene 17220 Mouse
Entrez Gene 288532 Rat
|The minichromosome maintenance (MCM7) helicase complex functions to initiate and elongate replication forks. Cell cycle checkpoint signaling pathways regulate DNA replication to maintain genomic stability (1). MCM7 is a putative human homologue of yeast CDC47 and a member of the MCM protein family, which has been implicated in the regulatory machinery causing DNA to replicate only once in the S phase. Findings indicate that MCM7 protein together with other MCM proteins participates in the regulation of mammalian DNA replication (2). In HeLa cells, depletion of MCM7 with small-interfering RNA suppressed ultraviolet (UV) light- or aphidicolin-induced hChk1 phosphorylation, and abolished UV-induced S-phase checkpoint activation. These results demonstrate that MCM7 plays a direct role in the transmission of DNA damage signals from active replication forks to the S-phase checkpoint machinery in human cells (3). |
|Transcription Factors DNA replicationCell cycle|
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Western blot - MCM7 antibody [EP1974Y]; Anti-MCM7 antibody [EP1974Y] at 1/10000 dilution + Hela cell lysate at 10 ug.Secondary.Goat anti-rabbit HRP labeled at 1/2000 dilution.Predicted band size : 81 kDa.Observed band size : 81 kDa.
Immunohistochemistry (Paraffin-embedded sections) - MCM7 antibody [EP1974Y]; Immunohistochemical analysis of paraffin-embedded human lung tissue using TA300952 at a 1/100 dilution.
Immunocytochemistry/ Immunofluorescence - MCM7 antibody [EP1974Y]; ICC/IF image of TA300952 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody , 1ug/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43uM.
Immunoprecipitation - Anti-MCM7 antibody [EP1974Y]; MCM7 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit monoclonal to MCM7 and 50ul 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 40ul SDS loading buffer and incubated for 10min at 70Â°C; 10ul of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with TA300952.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) .Band: 81kDa; MCM7
Flow Cytometry - MCM7 antibody [EP1974Y]; Overlay histogram showing HeLa cells stained with TA300952 (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 for 30 min at 22°C. The secondary antibody used was DyLight 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1ug/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed.