Anti msh2

Manufactured by Cell Signaling Technology

Sourced in France, United States

Anti-MSH2 is a primary antibody that targets the MSH2 protein. MSH2 is a key component of the DNA mismatch repair system and plays a critical role in maintaining genome stability. The Anti-MSH2 antibody can be used to detect and analyze the expression of the MSH2 protein in various experimental systems.

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Lab products found in correlation

Anti msh6, by Cell Signaling Technology (3 mentions) Anti α tubulin, by Merck Group (2 mentions) Anti mlh1, by Cell Signaling Technology (2 mentions) Anti γh2ax, by Merck Group (2 mentions) Anti dnmt1, by Merck Group (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Anti lamin b, by Santa Cruz Biotechnology (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti stat3, by Merck Group (1 mentions) Anti jak2, by Merck Group (1 mentions) Anti h3, by Merck Group (1 mentions) Anti ezh2, by Merck Group (1 mentions) Anti h3k27me3, by Merck Group (1 mentions) Anti bax, by Cell Signaling Technology (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti actin, by Merck Group (1 mentions) Anti bad, by Cell Signaling Technology (1 mentions) Anti cleaved parp, by Cell Signaling Technology (1 mentions) Anti caspase 3, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Anti-MSH2 #D24B5 (2 citations)

7 protocols using anti msh2

Co-IP and Western Blot of JAK2-Related Proteins

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For Co-IP of endogenous proteins, anti-JAK2 (Millipore, 06-255-I¸1:100), anti-MSH2 (Cell Signaling Technology (CST, MA), 2017S, 1:200), anti-MSH6 (Becton Dickinson (BD, NJ), 610918, 1:100), anti-EZH2 (CST, MA, 5246T, 1:200) antibodies were used. For western blot, anti-JAK2 (Millipore, 06-255-I, 1:1000 and Abcam, CA, ab108596, 1:1000 ), anti-p-JAK2 (Y1007/1008, 1:1000) (Millipore, MA, 07-606), anti-p-STAT3 (Y705) (CST, MA, 9145S, 1:1000), anti-LaminB (Santa Cruz (SC, CA), sc-6216, 1:1000), anti-GAPDH (CST, MA, 5174, 1:1000), anti-MSH2 (CST, MA, 2017S, 1:1000), anti-MSH6 (BD, 610918, 1:1000), anti-DNMT1 (Sigma, MO, D4692, 1:1000), anti-EZH2 (CST, MA, 3147S, 1:1000), anti-STAT3 (CST, MA, 9139, 1:1000), anti-γH2AX (CST, MA, 9718, 1:1000), anti-H3K27me3 (CST, MA, 9733, 1:1000), anti-H3 (CST, MA, 9751, 1:3000), anti-EZH2 (CST, MA, 3147S, 1:1000), anti-SUZ12 (CST, MA, 3737, 1:1000), anti-EED (Sigma, MO, GW10896A, 1:500) antibodies were used. For immunofluorescence, anti-JAK2 (Abcam, CA, ab108596, 1:100), anti-DNMT1 (SC, CA, sc-20701, 1:100), anti-γH2AX (Millipore, NJ, 05-636, 1:100), secondary Alexa Conjugate (CST, MA, mouse 8890, 1:500 and rabbit 8889, 1:1000) were used.

Ding N., Miller S.A., Savant S.S, & O’Hagan H.M. (2019). JAK2 regulates mismatch repair protein-mediated epigenetic alterations in response to oxidative damage. Environmental and molecular mutagenesis, 60(4), 308-319.

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Western Blot Quantification of Protein Biomarkers

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Cells were collected by centrifugation for GS-lines and by scraping for adherent cells. Westerns were done as described [23 (link)] and probed with respective antibodies: anti-α-Tubulin (Sigma, T5168, 1:10,000), anti- β-Actin (Bioconcept, 8H10D10. 1:10,000), anti-MGMT (R&D systems, AF3794-SP, 1:4000), anti-MSH6 (Cell Signaling, #5424 S, 1:4000), anti-MSH2 (Cell Signaling, #2017S, 1:4000). Membranes were washed 5 min x3 in TBS-T at RT, followed by incubation at RT for 1 h with the following secondary antibodies (according to primary antibody specifics): anti-rabbit (Promega, W4011, 1:5000), anti-mouse (Thermofisher, 31430, 1:5000), anti-goat (Thermofisher, 31402, 1:5000).

Tancredi A., Gusyatiner O., Bady P., Buri M.C., Lomazzi R., Chiesi D., Messerer M, & Hegi M.E. (2022). BET protein inhibition sensitizes glioblastoma cells to temozolomide treatment by attenuating MGMT expression. Cell Death & Disease, 13(12), 1037.

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Western Blot Analysis of DNA Repair Proteins

Cited 1 time

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Western blot analysis was performed as previously described.³⁶ (link) The following antibodies were used: anti-MLH1, anti-MSH2, anti-MSH6, anti-PMS2, anti-caspase-3, anti-cleaved caspase-3, anti-PARP, anti-cleaved PARP, anti-BAD, anti-pBAD (S112), anti-Bcl2, anti-Bax (Cell Signaling Technology), and anti-actin (Sigma).

Ye L., Jiang T., Shao H., Zhong L., Wang Z., Liu Y., Tang H., Qin B., Zhang X, & Fan J. (2017). miR-1290 Is a Biomarker in DNA-Mismatch-Repair-Deficient Colon Cancer and Promotes Resistance to 5-Fluorouracil by Directly Targeting hMSH2. Molecular Therapy. Nucleic Acids, 7, 453-464.

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Profiling DNA Damage Response Proteins

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Anti‐tyrosine‐701‐phosphorylated‐STAT1 (#9167), anti‐STAT3 (#4904), anti‐MLH1 (#4256), anti‐ATM (#2873), anti‐serine‐1981‐phosphorylated‐ATM (#5883), anti‐CHK2 (#6334), anti‐threonine‐68‐phosphorylated‐CHK2 (#2197), anti‐ATR (#13934), anti‐serine‐428‐phosphorylated‐ATR (#2853), anti‐CHK1 (#2360), anti‐serine‐345‐phosphorylated‐CHK1, anti‐HSP90 (#4874), anti‐serine‐139‐phosphorylated‐H2AX (#9718), anti‐c‐ABL(#2862), anti‐p53 (#9282), anti‐serine‐15‐phosphorylated‐p53 (#9284) and anti‐MSH2 (#2017) were purchased from Cell Signaling Technology (Ozyme, Montigny‐le‐Bretonneux, France). Anti‐STAT1α (#SC‐345), anti‐STAT1α and STAT1β isoforms (#SC‐346) and anti‐α‐actin (#SC‐1616) were from Santa Cruz/Tebu‐Bio (Le‐Perray‐en‐Yvelynes, France). Anti‐α‐tubulin (#T8203) was from Sigma‐Aldrich (Saint‐Quentin‐Fallavier, France).

Ah‐Koon L., Lesage D., Lemadre E., Souissi I., Fagard R., Varin‐Blank N., Fabre E.E, & Schischmanoff O. (2016). Cellular response to alkylating agent MNNG is impaired in STAT1‐deficients cells. Journal of Cellular and Molecular Medicine, 20(10), 1956-1965.

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Western Blot Analysis of Histone and DNA Repair Proteins

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Cells were lysed in RIPA lysis buffer (ThermoFisher), supplemented with PhosSTOP and cOmplete inhibitors (Sigma-Aldrich). Protein concentrations were measured utilizing the Pierce BCA Protein Assay (ThermoFisher). Samples were resolved on Bolt 4–12% Bis-Tris gels (ThermoFisher) and transferred to 0.2 µm nitrocellulose. Membranes were blocked in a 5% BSA/TBS-T solution for 1 h and then incubated with the following primary antibodies in blocking solution for 24 h at 4 °C: anti-GAPDH (1:2500), Cell Signaling, Cat#97166S), anti-β-Actin (1:5000, Cell Signaling, Cat#3700S), Histone H3 (acetyl K9, K14, K18, K23, K27) (1:1000, Abcam, Cat#ab47915), anti-PMS2 (1:1000, Abcam,Cat#ab11068), anti-MSH2 (1:1000, Cell Signaling, Cat#2017S), or anti-cleaved PARP (1:500, Cell Signaling, Cat#9541S). Secondary antibodies were used at 1:10,000 (Li-Cor IRDye 800CW Donkey anti-Mouse IgG and IRDye 680RD Goat anti-Rabbit IgG, Cat#926-32212 and Cat#926-68071, respectively).

Noll A., Myers C., Biery M.C., Meechan M., Tahiri S., Rajendran A., Berens M.E., Paine D., Byron S., Zhang J., Winter C., Pakiam F., Leary S.E., Cole B.L., Jackson E.R., Dun M.D., Foster J.B., Evans M.K., Pattwell S.S., Olson J.M, & Vitanza N.A. (2023). Therapeutic HDAC inhibition in hypermutant diffuse intrinsic pontine glioma. Neoplasia (New York, N.Y.), 43, 100921.

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Coimmunoprecipitation and Biotin-based Purification

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All coimmunoprecipitation and immunoprecipitation experiments were performed as previously described using 1000 μg of lysate in Cell Lytic M (Sigma Aldrich, Oakville, Ontario, Canada).^{4 (link)} No-antibody controls were included, although not always shown. Biotin-based protein purification was performed as previously described using 500 μg of lysate in RIPA buffer.^{18 (link)} These experiments were repeated at least three times. The following antibodies were used: anti-MSH2 (EMD, Billerica, MA, USA, IP/co-IP, catalog number NA27), anti-MSH6 (BD Biosciences, San Jose, CA, USA, catalog number 610919), anti-MSH2, anti-phospho-tyrosine, anti-ALK, anti-total caspase 3 (Cell Signaling Technologies, Danvers, MA, USA, catalog numbers 2017, 9416, 3633 and 14220, respectively) and anti-β-actin (Santa Cruz Biotechnology, Dallas, TX, USA; sc-47778).

Bone K.M., Wang P., Wu F., Wu C., Li L., Bacani J.T., Andrew S.E, & Lai R. (2015). NPM-ALK mediates phosphorylation of MSH2 at tyrosine 238, creating a functional deficiency in MSH2 and the loss of mismatch repair. Blood Cancer Journal, 5(5), e311-.

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Western Blot Analysis of DNA Mismatch Repair Proteins

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Cell pellets were lysed in 20 mM Tris (pH 8), 200 mM NaCl, 1 mM EDTA, 0.5% (v/v) NP40, 10% glycerol, supplemented with protease inhibitors. For western blotting, lysates were electrophoresed on Novex precast gels (Invitrogen) and immunoblotted using the following antibodies: anti-MSH6 (#5424), anti-MSH2 (#2017), anti-MLH1 (#4256), anti-thymidylate synthase (#9045), β-Actin (#4970), purchased from Cell Signaling. The following antibodies were also used; anti-MSH3 (sc-11441; Santa Cruz) and anti β-tubulin (T8328; Sigma). This was followed by incubation with anti-IgG-horseradish peroxidase and chemiluminescent detection (Supersignal West Pico Chemiluminescent Substrate, Pierce). Immunoblotting for β-Actin and β-tubulin were performed as loading control.

Guillotin D., Austin P., Begum R., Freitas M.O., Merve A., Brend T., Short S., Marino S, & Martin S.A. (2016). Drug-repositioning screens identify Triamterene as a selective drug for the treatment of DNA Mismatch Repair deficient cells. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(11), 2880-2890.

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