MCM2 antibody | CRCT2.1
Mouse anti Human MCM2 antibody, clone CRCT2.1 used for the evaluation of MCM2 expressionin treated HeLa cell lysates by western blotting.
Image caption:
Confirmation and in vitro characterization of Hit compounds. A) HeLa cells were treated with the ten compounds that produced the strongest reduction in pSer40/41MCM2 levels in the primary screen. Protein extracts were prepared and levels of residual pSer40/41MCM2 phosphorylation were measured by western blot analysis with the indicated antibodies. B) The same compounds were tested for their ability to inhibit CDC7 kinase activity in in vitro kinase assay. Kinase reactions were performed on a synthetic MCM2 substrate in the absence or presence of the indicated drug and reactions were run on SDS-PAGE gels. CDC7 activity on the synthetic MCM2 substrate was monitored by Western blotting with an anti-pSer108MCM2 antibody. As a control the levels of CDC7 kinase and synthetic MCM2 substrate present in each reaction were also assessed and shown to be similar in all reactions.
From: FitzGerald J, Murillo LS, O'Brien G, O'Connell E, O'Connor A, et al. (2014)
A High Through-Put Screen for Small Molecules Modulating MCM2 Phosphorylation Identifies Ryuvidine as an Inducer of the DNA Damage Response.
PLoS ONE 9(6): e98891.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Mouse anti Human MCM2 antibody, clone CRCT2.1 used for the evaluation of MCM2 expressionin treated HeLa cell lysates by western blotting.
Image caption:
Characterization of pSer40/41MCM2 antibody. A) HeLa whole cell extract was incubated with Lambda phosphatase in the presence or absence of phosphatase inhibitors. Proteins were analyzed by western blot with the indicated antibodies. B) Equal amounts of protein extract were separated on a single SDS-PAGE gel and transferred onto membranes. Vertical slices of membrane (each with identical protein content) were then incubated with anti-pSer40/41MCM2 antibody in the presence of the indicated competitor peptides (upper panels). Membranes were then re-probed with an anti-MCM2 antibody (lower panels). Identical exposures are shown. C) HeLa cells growing on coverslips were fixed and stained with anti-pSer40/41MCM2 antibody in the presence of the indicated competitor peptides. DNA was counterstained with DAPI. D) Serial sections of normal and cancerous colon tissue were used in IHC with the pSer40/41MCM2 antibody in the presence or absence of competitor peptide. Nuclear positivity is shown by brown color. E) HeLa cells were treated with PHA-767491 for the indicated time. Protein extracts were prepared and analysed by western blot with the indicated antibodies.
From: FitzGerald J, Murillo LS, O'Brien G, O'Connell E, O'Connor A, et al. (2014)
A High Through-Put Screen for Small Molecules Modulating MCM2 Phosphorylation Identifies Ryuvidine as an Inducer of the DNA Damage Response.
PLoS ONE 9(6): e98891.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Mouse anti Human MCM2 antibody, clone CRCT2.1 used for the evaluation of MCM2 expressionin treated HeLa cell lysates by western blotting.
Image caption:
Ryuvidine and Mitoxantrone reduce CDC7 levels and pSer40/41 MCM2 phosphorylation. A) Molecular structure of PHA-767491, Ryuvidine and Mitoxantrone. B) HeLa cells were incubated with either CDC7 inhibitor PHA-767491, Ryuvidine or Mitoxantrone at 10 micromolar each for the indicated time. Protein extracts were prepared and analyzed by western blot with the indicated antibodies. C) Levels of Cdc7-independent pSer41MCM2 and Cdc7-dependent pSer40/41MCM2 phosphorylation assessed in HeLa cells treated with the indicated concentration of PHA-767491, Ryuvidine or Mitoxantrone. D) Cells were incubated with either PHA-767491, Ryuvidine or Mitoxantrone at the indicated concentration for nine hours. Protein extracts were then prepared and analyzed by western blot with the indicated antibodies. E) HeLa cells incubated with either PHA-767491, Ryuvidine or Mitoxantrone at 10 micromolar each for nine hours in the presence or absence of 20 micromolar of caspase inhibitor Boc-D-fmk. Levels of Ser40/41MCM2 phosphorylation, CDC7 and MCM2 abundance as well as PARP cleavage, were assessed by western blotting.
From: FitzGerald J, Murillo LS, O'Brien G, O'Connell E, O'Connor A, et al. (2014)
A High Through-Put Screen for Small Molecules Modulating MCM2 Phosphorylation Identifies Ryuvidine as an Inducer of the DNA Damage Response.
PLoS ONE 9(6): e98891.
This image is from an open access article distributed under the terms of the Creative Commons Attribution License.
Mouse anti Human MCM2
- Product Type
- Monoclonal Antibody
- Clone
- CRCT2.1
- Isotype
- IgG1
- Specificity
- MCM2
Antibody Quality and Performance Guarantee
| Mouse anti Human MCM2 antibody, clone CRCT2.1 recognizes human DNA replication licensing factor MCM2, also known as Mcm2, Nuclear protein BM28 or mini chromosome maintenance protein-2. Mcm-2 is a 904 amino acid ~125kDa nuclear protein involved in the control of DNA replication. Mouse anti Human MCM2 antibody, clone CRCT2.1 has been used successfully for the detection of human MCM2 in ovarian adenocarcinoma by immunohistochemistry on formalin fixed, paraffin embedded tissues (Gakiopoulou et al. 2007). |
- Target Species
- Human
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% sodium azide (NaN3)
- Carrier Free
- Yes
- Immunogen
- Bacterially expressed human Mcm2.
- Approx. Protein Concentrations
- IgG concentration 1.0mg/ml
- Fusion Partners
- Spleen cells from immunised CBA/c F1 mice were fused with cells of the mouse SP-2/0 myeloma cell line.
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
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 visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Immunohistology - Paraffin 1 |  |
1/200 | |
| Western Blotting | |
- 1This product requires antigen retrieval using heat treatment prior to staining of paraffin sections.Sodium citrate buffer pH 6.0 is recommended for this purpose.
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.
- Histology Positive Control Tissue
- Human tonsil, or other tissue with high proliferative index
References for MCM2 antibody
-
(2011) Human Protein Atlas: CAB000303
Human Protein Atlas: CAB000303 -
Korkolopoulou, P. et al. (2005) Minichromosome maintenance proteins 2 and 5 expression in muscle-invasive urothelial cancer: a multivariate survival study including proliferation markers and cell cycle regulators.
Hum Pathol. 36: 899-907. -
Gakiopoulou, H. et al. (2007) Minichromosome maintenance proteins 2 and 5 in non-benign epithelial ovarian tumours: relationship with cell cycle regulators and prognostic implications.
Br J Cancer. 97:1124-34. -
Giaginis, C. et al. (2009) Clinical significance of MCM-2 and MCM-5 expression in colon cancer: association with clinicopathological parameters and tumor proliferative capacity.
Dig Dis Sci. 54: 282-91. -
Polo, S.E. et al. (2010) Clinical significance and prognostic value of chromatin assembly factor-1 overexpression in human solid tumours.
Histopathology. 57: 716-24. -
FitzGerald, J. et al. (2014) A high through-put screen for small molecules modulating MCM2 phosphorylation identifies Ryuvidine as an inducer of the DNA damage response.
PLoS One. 9: e98891. -
Montes de Oca, R. et al. (2015) The histone chaperone HJURP is a new independent prognostic marker for luminal A breast carcinoma.
Mol Oncol. 9 (3): 657-74. -
Wu, K.Z. et al. (2016) DDK dependent regulation of TOP2A at centromeres revealed by a chemical genetics approach.
Nucleic Acids Res. 44 (18): 8786-8798.
View The Latest Product References
- RRID
- AB_322519
- UniProt
- P49736
- Entrez Gene
- MCM2
- GO Terms
- GO:0000075 cell cycle checkpoint
- GO:0000082 G1/S transition of mitotic cell cycle
- GO:0000084 S phase of mitotic cell cycle
- GO:0005515 protein binding
- GO:0000216 M/G1 transition of mitotic cell cycle
- GO:0000785 chromatin
- GO:0046872 metal ion binding
- GO:0005524 ATP binding
- GO:0005654 nucleoplasm
- View More GO Terms
- GO:0006270 DNA-dependent DNA replication initiation
- GO:0006271 DNA strand elongation involved in DNA replication
- GO:0006350 transcription
- GO:0045449 regulation of transcription
MCA1859
0110R
1805
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