Revision 1

#89884Store at -20C

1 个试剂盒

(5 x 20 microliters)

Cell Signaling Technology

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

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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.

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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
    使用 MSH2 (D24B5) XP® Rabbit mAb 对 HeLa 和 NIH/3T3 细胞的提取物进行蛋白质印迹分析。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 2 Expand Image
    使用 PMS2 (E9U4P) Rabbit mAb(上图)和 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对各种人类细胞系提取物进行蛋白质印迹分析。OV-90 和 HCT 116 细胞中 PMS2 蛋白的低表达与预测的表达模式一致。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 3 Expand Image
    使用 MLH1 (D38G9) Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹分析。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 4 Expand Image
    使用 MSH6 (D60G2) XP® Rabbit mAb 对不同细胞类型的提取物进行蛋白质印迹法分析。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 5 Expand Image
    使用 EXO1 (E6P2B) Rabbit mAb(上图)或 GAPDH (D16H11) XP® Rabbit mAb #5174(下图)对来自各种细胞系的提取物进行蛋白质印迹分析。EXO1 蛋白在 Hs 822.T 细胞和 MBT2 细胞中低表达/阴性表达与预测的表达模式一致。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 6 Expand Image
    一抗与靶标蛋白结合之后,与偶联 HRP 的二抗形成复合体。添加 LumiGLO®,在酶催化分解期间发光。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 7 Expand Image
    在对照物肽(左)或抗原特异性肽(右)存在的情况下,使用 MSH2 (D24B5) XP® Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 8 Expand Image
    对 KYSE-70 细胞提取物的 PMS2 蛋白进行免疫沉淀分析。泳道 1 是 10% input,泳道 2 是 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 是 PMS2 (E9U4P) Rabbit mAb。使用 PMS2 兔单克隆抗体进行蛋白质印迹分析,该抗体检测的抗原决定簇与 PMS2 (E9U4P) Rabbit mAb 检测的抗原决定簇不同。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 9 Expand Image
    使用 MLH1 (D38G9) Rabbit mAb 对 HeLa 细胞中的 MLH1 进行免疫沉淀。使用同一抗体进行蛋白质进行印迹分析。IP () 为 5% Input。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 10 Expand Image
    使用 MSH6 (D60G2) XP® Rabbit mAb(绿色)对 293 细胞进行共聚焦免疫荧光分析。使用 DY-554 phalloidin(红色)标记肌动蛋白纤丝。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 11 Expand Image
    使用 MSH2 (D24B5) XP® Rabbit mAb(绿色),对 HeLa 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 DY-554 Phalloidin 进行标记(红色)。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 12 Expand Image
    使用 PMS2 (E9U4P) Rabbit mAb(绿色)和 MLH1 (4C9C7) Mouse mAb #3515(红色)对 KYSE-70 细胞(左图,阳性)和 HCT 116 细胞(右图,阴性)进行共聚焦免疫荧光分析。PMS2 与 MLH1 形成异二聚体(6)。肌动蛋白丝用 DyLight 554 Phalloidin #13054(伪彩青色)标记。蓝色 = DAPI #4083(DNA 荧光染料)。
    Mismatch DNA Repair (MMR) Antibody Sampler Kit: Image 13 Expand Image
    使用与非特异性阴性对照抗体(红)作对比的 MSH2 (D24B5) XP® Rabbit mAb 抗体(蓝),对 Hela 细胞进行流式细胞分析。