AHP1523 specifically recognises the nuclear protein MCM2 (mini chromosome maintenance protein 2), when phosphorylated at Ser27.
MCM2 is a putative DNA replicative helicase and member of the MCM family, critical during the initiation of DNA replication. During the G1 phase of the cell cycle the MCM proteins, part of the pre-replicative complex which assembles at replication origins, undergo phosphorylation by kinases including cyclin dependent kinase (CDK) and Cdc7/Dbf4 (DDK). Studies have shown that the phosphorylation of MCM2 occurs on multiple sites, namely Ser27, Ser41, Ser53, Ser108 and Ser139 and that phosphorylation results in a conformational change of the complex and activation of the helicase activity.
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
Western Blotting(1)
1/10000 - 1/20000
(1)
For the detection of phosphoproteins, serine phosphatase inhibitors such as 10mM Sodium Fluoride should be added to the sample buffer. Milk or other casein-based blocking solutions are not recommended as casein is a phosphoprotein and its use can result in high background.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
AHP1523 detects a band of approximately 120-130kDa in HeLa cell lysates
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Antiserum Preparation
Antiserum to human MCM2 was raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
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