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MCM5 Recombinant Rabbit mAb

The Hela (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-MCM5 antibody (AP94436,1:100); Isotype Control (orange): Rabbit IgG (AP94436). Blank control (black): PBS. Acquisition of 20,000 events was performed.
25 ug total protein per lane of various lysates (see on figure) probed with MCM5 monoclonal antibody, unconjugated (AP94436) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with MCM5 Monoclonal Antibody, Unconjugated (AP94436) at 1:200 overnight at 4°C, followed by conjugation to the AP94436-HRP and DAB (C-0010) staining.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IHC-F, IF
Reactivity	Rat, Human, Mouse
Host	Rabbit
Clonality	Recombinant
Physical State	Liquid
Isotype	IgG/Kappa
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Nucleus.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	The mini-chromosome maintenance (MCM) family of proteins, including MCM2, MCM3, MCM4 (Cdc21), MCM5 (Cdc46), MCM6 (Mis5) and MCM7 (Cdc47), are regulators of DNA replication that act to ensure replication occurs only once in the cell cycle. Expression of MCM proteins increases during cell growth, peaking at G1 to S phase. The MCM proteins each contain an ATP-binding motif, which is predicted to mediate ATP-dependent opening of double-stranded DNA. MCM proteins are regulated by E2F transcription factors, which induce MCM expression, and by protein kinases, which interact with MCM proteins to maintain the postreplicative state of the cell. MCM2/MCM4 complexes function as substrates for Cdc2/cyclin B in vitro. Cleavage of MCM3, which can be prevented by caspase inhibitors, results in the inactivation during apoptosis of the MCM complex, which is composed of, at least, MCM2–6. A complex composed of MCM4, MCM6 and MCM7 has been shown to be involved in DNA helicase activity, and MCM5 is involved in IFN-induced Stat1 Alpha transcription activation.

Additional Information
Dilution	WB=1:500-2000,IHC-P=1:50-200,IHC-F=1:50-200,IF=1:50-200,Flow-Cyt=1:50-100
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

For Research Use Only. Not For Use In Diagnostic Procedures.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol