Revision 1

#89884Store at -20C

1 Kit

(5 x 20 microliters)

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.
Product Includes Product # Quantity Mol. Wt Isotype/Source
MSH2 (D24B5) XP® Rabbit mAb 2017 20 µl 100 kDa Rabbit IgG
MLH1 (D38G9) Rabbit mAb 4256 20 µl 84 kDa Rabbit IgG
MSH6 (D60G2) XP® Rabbit mAb 5424 20 µl 160 kDa Rabbit IgG
PMS2 (E9U4P) Rabbit mAb 27884 20 µl 110 kDa Rabbit IgG
EXO1 (E6P2B) Rabbit mAb 63862 20 µl 120 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Mismatch DNA Repair (MMR) Antibody Sampler Kit provides an economical means of detecting proteins involved in MMR. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

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.

Background

DNA mismatch repair (MMR), a conserved process for detecting and correcting errors made during DNA synthesis, is crucial to the maintenance of genomic integrity (1). In prokaryotes, a MutS homodimer recruits a MutL homodimer to sites of DNA mismatches. In eukaryotes, six MutS homologues (MSH1-6) and four MutL homologues (MLH1, PMS2, PMS1, and MLH3) have been identified. Heterodimers composed of two MutL homologues detect distinct DNA mismatch lesions, and heterodimers composed of two MutS homologues perform the repair (2). Other factors required for MMR in eukaryotes are EXO1, PCNA, RFC, RPA, DNA polymerases, and DNA ligases (1).

Microsatellite instability (MSI) is a predisposition to genetic mutation resulting from MMR deficiency (dMMR). High MSI (MSI-H) arising from dMMR results in Lynch syndrome, also known as hereditary non-polyposis colorectal cancer (HNPCC). Lynch syndrome is associated with colon cancer, as well as other human cancers (3). MSI and dMMR are strongly associated with tumor responsiveness to immune checkpoint blockade (4,5). MSI status can be determined through PCR amplification of microsatellite markers and/or immunohistochemical detection of MMR proteins MLH1, PMS2, MSH2, and MSH6. The absence of expression of any of these MMR proteins indicates dMMR (3).

  1. Modrich, P. (2006) J Biol Chem 281, 30305-9.
  2. Kolodner, R.D. and Marsischky, G.T. (1999) Curr Opin Genet Dev 9, 89-96.
  3. Pećina-Šlaus, N. et al. (2020) Front Mol Biosci 7, 122.
  4. Kok, M. et al. (2019) ESMO Open 4, e000511.
  5. Yi, M. et al. (2018) Mol Cancer 17, 129.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered 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

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    Revision 1
    #89884

    Mismatch DNA Repair (MMR) Antibody Sampler Kit

    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts of HeLa and NIH/3T3 cells using MSH2 (D24B5) XP® Rabbit mAb.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various human cell lines using PMS2 (E9U4P) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of PMS2 protein in OV-90 and HCT 116 cells is consistent with the predicted expression pattern.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell types using MLH1 (D38G9) Rabbit mAb.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell types using MSH6 (D60G2) XP® Rabbit mAb.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using EXO1 (E6P2B) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low/negative expression of EXO1 protein in Hs 822.T and MBT2 cells is consistent with the predicted expression pattern.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 6 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 7 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma using MSH2 (D24B5) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 8 Expand Image
    Immunoprecipitation of PMS2 protein from KYSE-70 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is PMS2 (E9U4P) Rabbit mAb. Western blot analysis was performed using a PMS2 rabbit monoclonal antibody which detects an epitope distinct from that detected by PMS2 (E9U4P) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody. 
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 9 Expand Image
    Immunoprecipitation of MLH1 from HeLa cells using MLH1 (D38G9) Rabbit mAb. Western blot was performed using the same antibody. IP () is 5% input.
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of 293 cells using MSH6 (D60G2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 11 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using MSH2 (D24B5) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 12 Expand Image
    Confocal immunofluorescent analysis of KYSE-70 cells (left, positive) and HCT 116 cells (right, negative) using PMS2 (E9U4P) Rabbit mAb (green) and MLH1 (4C9C7) Mouse mAb #3515 (red). PMS2 forms a heterodimer with MLH1 (6). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (cyan pseudocolor). Blue = DAPI #4083 (fluorescent DNA dye).
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 13 Expand Image
    Flow cytometric analysis of HeLa cells using MSH2 (D24B5) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).