MCM-2

MCM-2 (mini-chromosome maintenance 2) is a human gene. The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that is involved in the initiation of eukaryotic genome replication. The hexameric protein complex formed by MCM proteins is a key component of the pre-replication complex, and may be involved in the formation of replication forks and in the recruitment of other DNA replicationrelated proteins. This protein forms a complex with MCM-4, 6, and 7, and has been shown to regulate the helicase activity of the complex. This protein is phosphorylated, and thus regulated by protein kinases CDC2 and CDC7.

MCM-2 is essential for eukaryotic DNA replication and drives the formation of pre-replicative complexes, which is the key first step during the G1 phase. Therefore, altered MCM-2 expression may be a hallmark of cell-cycle deregulation, which could be the most essential mechanism in the development and progression of human cancers. MCM2 has been identified by DNA microarray and transcriptional profiling as a gene that is overexpressed in Cervical Carcinomas. This protein is over-expressed in Cervical Dysplasia as a result of HPV infection. The over-expression of MCM-2 provides the link between oncogenic HPV infection and the molecular event of Cervical Dysplasia.

MCM2 is a rabbit monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

1. Murphy N, et al. J Clin Pathol. 2005;58:525-534
2. Chen Y, et al. Cancer Res. 2003;63:1927-1935
3. Santin AD, et al. Virology. 2005;331:269-291
4. Lei M, Curr Cancer Drug Targets. 2005;5:365-80
5. Ishimi Y, et al. Eur J Biochem. 2003;270:1089-101

1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
5. Any of three heating methods may be used:
   • Electric Pressure Cooker
     Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
   • Water Bath Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
   • Conventional Steamer Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.