MCM2 polyclonal antibody
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More Files
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Western Blot (Cell lysate)
Western blot using MCM2 polyclonal antibody (Cat # PAB10264) shows detection of both phosphorylated and unphosphorylated MCM2 presentin nuclear extracts from elutriated human cells (MO59K/K-562).
The MCM2 protein is phosphorylated after initiation of DNA replication, therefore, the proteinis unphosphorylated in early S phase, and gradually becomes phosphorylated throughout S phase.
In G2/M, all MCM2 is phosphorylated.
Panel A shows western blot results for lysates were prepared from asynchronous cells (Lane 1), cells arrested in early S with aphidicolin (lane2), and cells arrested in mitosis with nocodazole (Lane 3).
Panel B shows a schematic diagram of bands representing phosphorylated and unphosphorylated MCM2 present in these preparations.
Asynchronous cells contain a doublet of both forms.
Aphidicolin treated cells contain only unphosphorylated MCM2 and nocodozole treatment results in only phosphorylated MCM2 detected in the lysate.
The phosphorylated band migrates faster than the unphosphorylated form and isseen as the lower band.
There is a clear switch from the unphosphorylated form in the center lane, to the phosphorylated form in the third lane, confirming recognition of both forms of MCM2 by this antibody.
The primary antibody was diluted 1:400 for this experiment.
Personal Communication, Jennifer Seiler,NIH CCR Bethesda MD.
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Specification
Product Description
Rabbit polyclonal antibody raised against synthetic peptide of MCM2.
Immunogen
A synthetic peptide corresponding to amino acids 21-31 of human MCM2.
Host
Rabbit
Reactivity
Human, Mouse, Rat, Yeast
Specificity
This affinity-purified antibody is directed against human MCM2 protein. Approximately equivalent reactivity occurs against both unphosphorylated and phosphorylated forms of human MCM2 (phosphorylated at residues pS26 and pS27).
Form
Liquid
Quality Control Testing
Antibody Reactive Against Synthetic Peptide.
Recommend Usage
ELISA (1:10000-1:50000)
Western Blot (1:500-1:2000)
The optimal working dilution should be determined by the end user.Storage Buffer
In 20 mM KH2PO4, 150 mM NaCl, pH 7.2 (0.01% sodium azide)
Storage Instruction
Store at 4°C. For long term storage store at -20°C.
Aliquot to avoid repeated freezing and thawing.Note
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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Applications
Western Blot (Cell lysate)
Western blot using MCM2 polyclonal antibody (Cat # PAB10264) shows detection of both phosphorylated and unphosphorylated MCM2 presentin nuclear extracts from elutriated human cells (MO59K/K-562).
The MCM2 protein is phosphorylated after initiation of DNA replication, therefore, the proteinis unphosphorylated in early S phase, and gradually becomes phosphorylated throughout S phase.
In G2/M, all MCM2 is phosphorylated.
Panel A shows western blot results for lysates were prepared from asynchronous cells (Lane 1), cells arrested in early S with aphidicolin (lane2), and cells arrested in mitosis with nocodazole (Lane 3).
Panel B shows a schematic diagram of bands representing phosphorylated and unphosphorylated MCM2 present in these preparations.
Asynchronous cells contain a doublet of both forms.
Aphidicolin treated cells contain only unphosphorylated MCM2 and nocodozole treatment results in only phosphorylated MCM2 detected in the lysate.
The phosphorylated band migrates faster than the unphosphorylated form and isseen as the lower band.
There is a clear switch from the unphosphorylated form in the center lane, to the phosphorylated form in the third lane, confirming recognition of both forms of MCM2 by this antibody.
The primary antibody was diluted 1:400 for this experiment.
Personal Communication, Jennifer Seiler,NIH CCR Bethesda MD.Enzyme-linked Immunoabsorbent Assay
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Gene Info — MCM2
Entrez GeneID
4171Protein Accession#
NP_004517;P49736Gene Name
MCM2
Gene Alias
BM28, CCNL1, CDCL1, D3S3194, KIAA0030, MGC10606, MITOTIN, cdc19
Gene Description
minichromosome maintenance complex component 2
Omim ID
116945Gene Ontology
HyperlinkGene Summary
The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are 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 (pre_RC) and may be involved in the formation of replication forks and in the recruitment of other DNA replication related proteins. This protein forms a complex with MCM4, 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. [provided by RefSeq
Other Designations
DNA replication licensing factor MCM2|MCM2 minichromosome maintenance deficient 2, mitotin|cell devision cycle-like 1|cyclin-like 1|minichromosome maintenance deficient 2 (mitotin)|nuclear protein BM28
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Interactome
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Pathway
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Disease
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Publication Reference
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Interaction of hRad51 and hRad52 with MCM complex: a cross-talk between recombination and replication proteins.
Shukla A, Navadgi VM, Mallikarjuna K, Rao BJ.
Biochemical and Biophysical Research Communications 2005 Apr; 329(4):1240.
Application：WB-Ce, Human, HeLa cells.
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Interaction of chromatin-associated Plk1 and Mcm7.
Tsvetkov L, Stern DF.
The Journal of Biological Chemistry 2005 Mar; 280(12):11943.
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A human nuclear protein with sequence homology to a family of early S phase proteins is required for entry into S phase and for cell division.
Todorov IT, Pepperkok R, Philipova RN, Kearsey SE, Ansorge W, Werner D.
Journal of Cell Science 1994 Jan; 107(Pt1):253.
Application：IF, Microinjection, WB, Human, Mouse, HeLa, IMR90, NIH/3T3, PtK2 cells.
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Interaction of hRad51 and hRad52 with MCM complex: a cross-talk between recombination and replication proteins.
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