MCM2 Antibody | CRCT2.1

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MCM2 Antibody | CRCT2.1 gallery image 1

Paraffin embedded human tonsil stained with Mouse anti Human MCM2 (MCA1859) following citrate antigen retrieval

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MCM2 Antibody | CRCT2.1 gallery image 2

Published customer image:
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.

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Published customer imageCharacterization 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.

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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.

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  • Mouse anti Human MCM2
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  • Product Type
    Monoclonal Antibody
  • Clone
    CRCT2.1
  • Isotype
    IgG1
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA1859P *, WBdatasheet pdfdatasheet pdf0.1 mg
    MCA1859
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • 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).
    • 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 NameYesNoMin DilutionMax Dilution
      Western Blotting
      Immunohistology - Paraffin(1)1/200
      (1)
      This 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 antibody 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 antibody for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
      Tonsil, any tissue with high proliferative index
    • Immunofluorescence
    • Western Blotting
    • Instructions For Use

    Additional MCM2 Antibody Formats

    Formats Clone Applications Sizes available
    MCM2 Antibody : Purified CRCT2.1 P *, WB 0.1 mg
    • Copyright © 2016 Bio-Rad

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              Tonsil, any tissue with high proliferative index
              • Application NameReference Images
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              References

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