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Anti mst2

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Anti-Mst2 is a laboratory reagent produced by Cell Signaling Technology. It is a primary antibody that detects endogenous levels of total Mst2 protein. Mst2 is a serine/threonine kinase that plays a role in the regulation of the Hippo signaling pathway.

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Lab products found in correlation

Anti mst1, by Cell Signaling Technology (6 mentions) Ripa buffer, by Thermo Fisher Scientific (6 mentions) Anti yap, by Cell Signaling Technology (3 mentions) Anti lats1, by Cell Signaling Technology (2 mentions) Anti mob1, by Cell Signaling Technology (2 mentions) Complete protease inhibitor, by Merck Group (2 mentions) Odyssey imaging system, by LI COR (2 mentions) Anti actin, by Merck Group (2 mentions) Pvdf membrane, by Thermo Fisher Scientific (2 mentions) Anti phospho yap s127, by Cell Signaling Technology (1 mentions) Anti tubulin, by Merck Group (1 mentions) Anti ctcf, by Cell Signaling Technology (1 mentions) Anti flag, by Merck Group (1 mentions) Phosphatase inhibitor, by Merck Group (1 mentions) β actin sc 8432, by Santa Cruz Biotechnology (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti p lats1 t1079, by Cell Signaling Technology (1 mentions) Anti sav1, by Cell Signaling Technology (1 mentions) Anti pyap s127, by Cell Signaling Technology (1 mentions) Anti parp, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Rabbit anti-MST2 (2 citations) Anti-Phospho-MST1 (Thr183)/MST2 (Thr180) (3681 (2 citations)

7 protocols using anti mst2

1

Culturing Cancer Cell Lines

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The HEK293, MCF7 breast cancer cells, and A549 lung cancer cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). HCT116 colon cancer cells were cultured in McCoy’s 5A medium with 10% FBS. Cell detachment was achieved by culturing cells in poly–HEMA (2-hydroxyethyl methacrylate)–coated dishes. Where indicated, the following drugs were used: phosphatase inhibitors (10 mM sodium fluoride, 10 mM sodium pyrophosphate) and 2-DG (25 mM) were from Sigma. Glucose-free DMEM was from Thermo Fisher Scientific. The following antibodies were obtained commercially: anti-CTCF (Cell Signaling Technology, #3418), anti–phospho-RxxS/T motif (Cell Signaling Technology, #9614), anti-LATS1 (Cell Signaling Technology, #3477), anti-MST2 (Cell Signaling Technology, #3952), anti-YAP (Cell Signaling Technology, #14074), anti–phospho-YAP (S127) (Cell Signaling Technology, #13008), anti-Flag (Sigma, #F1804), and anti-tubulin (Sigma, #T9026). Rabbit polyclonal anti–phospho-CTCF (S402) antibodies were custom generated using a specific phosphopeptide (GenScript).
Luo H., Yu Q., Liu Y., Tang M., Liang M., Zhang D., Xiao T.S., Wu L., Tan M., Ruan Y., Bungert J, & Lu J. (2020). LATS kinase–mediated CTCF phosphorylation and selective loss of genomic binding. Science Advances, 6(8), eaaw4651.
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2

Hippo Signaling Pathway Antibodies

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Antibodies against caspase 9 (sc-56077) and β-actin (sc-8432) were obtained from Santa Cruz Biotechnology, Texas, United States. Others were purchased from Cell Signaling Technology: anti-PARP (#9542), anti-cleaved caspase 3 (#9661), anti-MST1 (#3682), anti-MST2 (#3952), anti-p-MST1(T183)/MST2(T180) (#49332), anti-LATS1 (#3477), anti-p-LATS1(S909) (#9159), anti-p-LATS1(T1079) (#8654), anti-MOB1 (#13730), anti-p-MOB1 (T35) (#8699), anti-SAV1 (#13301), anti-YAP (#14074), anti-p-YAP (S127) (#13008), and anti-p-YAP (S397) (#13619). RNase A and Propidium Iodide/PI were purchased from Coolaber (Beijing, China) and Sigma-Aldrich (United States), respectively.
Wang H., Wang R., Huang D., Li S., Gao B., Kang Z., Tang B., Xie J., Yan F., Liang R., Li H, & Yan J. (2021). Homoharringtonine Exerts Anti-tumor Effects in Hepatocellular Carcinoma Through Activation of the Hippo Pathway. Frontiers in Pharmacology, 12, 592071.
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3

Immunoblotting of Mst1 and Mst2

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Cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Thermo Fisher Scientific) containing cOmplete protease inhibitor (Sigma) and frozen. Equal amounts of protein were separated on 4%–12% bis-Tris NuPage gels and transferred to polyvinylidene fluoride (PVDF) membranes (Thermo Fisher Scientific). Immunodetection was performed using anti-Mst1 (Cell Signaling Technology cat# 3682S), anti-Mst2 (Cell Signaling Technology cat# 3952S), and anti-actin (Sigma cat# A3854). Proteins were visualized with the Odyssey imaging system (LI-COR Biosciences).
Cohen T.S., Boland M.L., Boland B.B., Takahashi V., Tovchigrechko A., Lee Y., Wilde A.D., Mazaitis M.J., Jones-Nelson O., Tkaczyk C., Raja R., Stover C.K, & Sellman B.R. (2018). S. aureus Evades Macrophage Killing through NLRP3-Dependent Effects on Mitochondrial Trafficking. Cell reports, 22(9), 2431-2441.
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4

Cell Line Authentication and Validation

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C4-2B4, DU145, LNCaP, PC3, HEK293, 293FT, and Phoenix cells were from American Type Culture Collection. PC3-mm2 was kindly provided by Dr. Isaiah Fidler (University of Texas, M. D. Anderson Cancer Center). All the cell lines were authenticated by short tandem repeat DNA profiling. Anti-YAP, anti-phospho-YAP (Ser127), anti-LATS1, anti-phospho-LATS1 (Thr1079), anti-MST1, anti-MST2, anti-ERK1/2, anti-phospho-ERK1/2 (Thr202/Tyr204), and anti-lamin A/C were from Cell Signaling Technology (Boston, MA).
Zeng H., Ortiz A., Shen P.F., Cheng C.J., Lee Y.C., Yu G., Lin S.C., Creighton C.J., Yu-Lee L.Y, & Lin S.H. (2016). Angiomotin regulates prostate cancer cell proliferation by signaling through the Hippo-YAP pathway. Oncotarget, 8(6), 10145-10160.
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5

Protein Expression Analysis in Liver Tissue

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The liver tissues were homogenized in RIPA buffer (Thermo Fisher) to harvest proteins. Total protein was resolved on TGX gradient gels (BioRad). The following primary antibodies were used: anti-β-actin (Sigma), anti-a-catenin (Cell Signaling), anti-cyclin D1 (Millipore), anti-Mst1 (Cell Signaling), anti-Mst2 (Cell Signaling), anti-NF2 (Sigma), anti-p21 (Santa Cruz), anti-pYap1 (Cell Signaling), anti-Yap1, phospho-pH2A.X. (Abcam) Licor Odyssey Imaging system was used to visualize protein bands. Gray scale images were quantified with ImageJ as described previously(24 (link)).
Yin H., Bogorad R.L., Barnes C., Walsha S., Zhuang I., Nonaka H., Ruda V., Kuchimanchi S., Nechev L., Akinc A., Xue W., Zerial M., Langer R., Anderson D.G, & Koteliansky V. (2015). Control of liver size by RNAi-mediated multiplex knockdown and its application for discovery of regulatory mechanisms. Journal of hepatology, 64(4), 899-907.
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6

Immunoblot Analysis of Cellular Signaling

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Gels for SDS-PAGE or Phos-tag SDS-PAGE were prepared according to the manufacturer’s instructions (NARD Institute). Separated proteins were transferred onto a PVDF membrane and then were identified by immunoblot analysis with the appropriate primary antibodies at a dilution of 1:1000 (or as otherwise stated below). Antibody to Mob1 phosphorylated at Thr35 (1:2000 dilution; 8699), anti-Mob1 (1:2000 dilution; 13730), anti-Mst1 (1:2000 dilution; 3682), anti-Mst2 (1:2000 dilution; 3952), anti-GAPDH (1:5000 dilution; 5174), antibody to phosphorylated p38 (1:3000 dilution; 9211), antibody to phosphorylated Jnk (1:3000 dilution; 4668), and anti-β-actin (1:5000 dilution; 8457) were from Cell Signaling Technology. Anti-Rac1 (66122-1-Ig), anti-HA Rabbit (51064-2-AP), and anti-Flag Rabbit (1:3000 dilution; 20543-1-AP) were from Proteintech. Anti-gamma Tubulin (1:5000 dilution; ab11316) was from Abcam and anti-Flag Mouse (1:5000 dilution; sc-166355) was from Santa Cruz Biotechnology. The protein bands were visualized with a SuperSignal West Pico Kit according to the manufacturer’s instructions (Thermo Fisher Scientific Pierce).
Geng J., Shi Y., Zhang J., Yang B., Wang P., Yuan W., Zhao H., Li J., Qin F., Hong L., Xie C., Deng X., Sun Y., Wu C., Chen L, & Zhou D. (2021). TLR4 signalling via Piezo1 engages and enhances the macrophage mediated host response during bacterial infection. Nature Communications, 12, 3519.
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7

Immunoblotting of Mst1 and Mst2

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Cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Thermo Fisher Scientific) containing cOmplete protease inhibitor (Sigma) and frozen. Equal amounts of protein were separated on 4%–12% bis-Tris NuPage gels and transferred to polyvinylidene fluoride (PVDF) membranes (Thermo Fisher Scientific). Immunodetection was performed using anti-Mst1 (Cell Signaling Technology cat# 3682S), anti-Mst2 (Cell Signaling Technology cat# 3952S), and anti-actin (Sigma cat# A3854). Proteins were visualized with the Odyssey imaging system (LI-COR Biosciences).
Cohen T.S., Boland M.L., Boland B.B., Takahashi V., Tovchigrechko A., Lee Y., Wilde A.D., Mazaitis M.J., Jones-Nelson O., Tkaczyk C., Raja R., Stover C.K, & Sellman B.R. (2018). S. aureus Evades Macrophage Killing through NLRP3-Dependent Effects on Mitochondrial Trafficking. Cell reports, 22(9), 2431-2441.
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