Phospho-MCM2 (Ser139) Antibody [D19G1]

Application: Reactivity: Human, Mouse, Rat, Hamster, Monkey, Dog

Lane 1: Hela, Lane 2: Hela (λ phosphatase and CIP treated)

Usage Information

Dilution
WB1:1000 IP1:50 IF1:100 FCM1:200

Application
WB, IP, IF, FCM

Reactivity
Human, Mouse, Rat, Hamster, Monkey, Dog

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
125 kDa

Datasheet & SDS

Biological Description

Specificity
Phospho-MCM2 (Ser139) Antibody [D19G1] detects endogenous levels of total MCM2 protein only when it is phosphorylated at Ser139.

Clone
D19G1

Synonym(s)
DNA replication licensing factor MCM2; MCM2

Background
Phospho‑MCM2 (Ser139) marks a key regulatory site on the MCM2 subunit of the MCM2‑7 replicative helicase, which licenses origins and drives DNA‑fork unwinding during S phase. MCM2 is part of a heterohexameric ring that assembles with MCM3–7 at origins as part of the pre‑replication complex, and phosphorylation of MCM2 at Ser139 by the Cdc7/Dbf4 kinase in late G1 and early S phase coincides with the activation of replication forks, linking origin firing to cell‑cycle progression. In mammalian cells, Cdc7‑dependent phosphorylation of Ser139, along with neighboring Ser27 and Ser41, enhances the ATPase activity of the MCM2‑7 ring, promotes chromatin‑bound MCM2 phosphorylation, and is required for the orderly initiation of DNA synthesis, while non‑phosphorylatable MCM2 variants impair replication initiation and reduce helicase efficiency. The MCM2‑7 complex additionally participates in replication‑fork stability and checkpoint responses by interacting with checkpoint and homologous‑recombination regulators, and its dynamic phosphorylation ensures that each origin fires only once per cycle and that stalled forks can be protected and restarted.

Background

Phospho‑MCM2 (Ser139) marks a key regulatory site on the MCM2 subunit of the MCM2‑7 replicative helicase, which licenses origins and drives DNA‑fork unwinding during S phase. MCM2 is part of a heterohexameric ring that assembles with MCM3–7 at origins as part of the pre‑replication complex, and phosphorylation of MCM2 at Ser139 by the Cdc7/Dbf4 kinase in late G1 and early S phase coincides with the activation of replication forks, linking origin firing to cell‑cycle progression. In mammalian cells, Cdc7‑dependent phosphorylation of Ser139, along with neighboring Ser27 and Ser41, enhances the ATPase activity of the MCM2‑7 ring, promotes chromatin‑bound MCM2 phosphorylation, and is required for the orderly initiation of DNA synthesis, while non‑phosphorylatable MCM2 variants impair replication initiation and reduce helicase efficiency. The MCM2‑7 complex additionally participates in replication‑fork stability and checkpoint responses by interacting with checkpoint and homologous‑recombination regulators, and its dynamic phosphorylation ensures that each origin fires only once per cycle and that stalled forks can be protected and restarted.

References
https://pubmed.ncbi.nlm.nih.gov/18180284/ https://pubmed.ncbi.nlm.nih.gov/16899510/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%