Revision 1
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP

REACTIVITY:

H M R Hm Mk Dg

SENSITIVITY:

Endogenous

MW (kDa):

125

Source/Isotype:

Rabbit IgG

UniProt ID:

#P49736

Entrez-Gene Id:

4171

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:100
Flow Cytometry (Fixed/Permeabilized) 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-MCM2 (Ser139) (D1Z8X) XP® recognizes endogenous levels of MCM2 protein only when phosphorylated at Ser139.

Species Reactivity:

Human, Mouse, Rat, Hamster, Monkey, Dog

Species predicted to react based on 100% sequence homology

Xenopus

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser139 of human MCM2 protein.

Background

The minichromosome maintenance (MCM) 2-7 proteins are a family of six related proteins required for initiation and elongation of DNA replication. MCM2-7 bind together to form the heterohexameric MCM complex that is thought to act as a replicative helicase at the DNA replication fork (1-5). This complex is a key component of the pre-replication complex (pre-RC) (reviewed in 1). Cdc6 and CDT1 recruit the MCM complex to the origin recognition complex (ORC) during late mitosis/early G1 phase forming the pre-RC and licensing the DNA for replication (reviewed in 2). Licensing of the chromatin permits the DNA to replicate only once per cell cycle, thereby helping to ensure that genetic alterations and malignant cell growth do not occur (reviewed in 3). Phosphorylation of the MCM2, MCM3, MCM4, and MCM6 subunits appears to regulate MCM complex activity and the initiation of DNA synthesis (6-8). CDK1 phosphorylation of MCM3 at Ser112 during late mitosis/early G1 phase has been shown to initiate complex formation and chromatin loading in vitro (8). Phosphorylation of MCM2 at serine 139 by cdc7/dbf4 coincides with the initiation of DNA replication (9). MCM proteins are removed during DNA replication, causing chromatin to become unlicensed through inhibition of pre-RC reformation. Studies have shown that the MCM complex is involved in checkpoint control by protecting the structure of the replication fork and assisting in restarting replication by recruiting checkpoint proteins after arrest (reviewed in 3,10).

  1. Lei, M. and Tye, B.K. (2001) J Cell Sci 114, 1447-54.
  2. Lygerou, Z. and Nurse, P. (2000) Science 290, 2271-3.
  3. Forsburg, S.L. (2004) Microbiol Mol Biol Rev 68, 109-31.
  4. Tye, B.K. and Sawyer, S. (2000) J Biol Chem 275, 34833-6.
  5. Labib, K. et al. (2000) Science 288, 1643-7.
  6. Charych, D.H. et al. (2008) J Cell Biochem 104, 1075-86.
  7. Masai, H. et al. (2006) J Biol Chem 281, 39249-61.
  8. Lin, D.I. et al. (2008) Proc Natl Acad Sci USA 105, 8079-84.
  9. Tsuji, T. et al. (2006) Mol Biol Cell 17, 4459-72.
  10. Bailis, J.M. et al. (2008) Mol Cell Biol 28, 1724-38.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

Revision 1
#12958

Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb

Western Blotting Image 1: Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb Expand Image
Western blot analysis of HeLa cell extracts, untreated (-) or calf intestinal phosphatase (CIP) and λ phosphatase-treated (+), and extracts of T98G cells serum-starved for 48 hours (-) or serum starved for 48 hours followed by serum-stimulation for 24 hours (+), using Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb (upper), MCM2 (D7G11) XP® Rabbit mAb #3619 or α-Actinin (D6F6) XP® Rabbit mAb #6487.
Immunoprecipitation Image 1: Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb Expand Image
Immunoprecipitation of phospho-MCM2 (Ser139) from T98G cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb.
Immunofluorescence Image 1: Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of WI-38 cells, serum-deprived (48 hr; upper left), or serum-treated (10%, 24 hr; upper right) and HeLa cells, untreated (lower left), or λ phosphatase-treated (2 hr; lower right), using Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow Cytometry Image 1: Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb Expand Image
Flow cytometric analysis of Jurkat cells using Phospho-MCM2 (Ser139) (D1Z8X) XP® Rabbit mAb and Propidium Iodide (PI)/RNase Staining Solution #4087 (DNA content). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 conjugate) #4412 was used as a secondary Ab.