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Anti phospho mypt1

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Anti-phospho-MYPT1 is a primary antibody product that detects phosphorylation of the regulatory subunit of myosin phosphatase (MYPT1). It is used to monitor the activity of MYPT1, a key regulator of myosin light chain phosphorylation and cellular contractility.

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Lab products found in correlation

Anti phospho mlc2, by Cell Signaling Technology (2 mentions) Anti mlc2, by Cell Signaling Technology (2 mentions) Anti rhoa, by Cell Signaling Technology (2 mentions) Anti cdc42, by Cell Signaling Technology (2 mentions) Anti pkcε, by Cell Signaling Technology (2 mentions) Anti e cadherin, by R&D Systems (2 mentions) Anti pkcε, by Santa Cruz Biotechnology (2 mentions) Anti phospho smad2 3, by Cell Signaling Technology (2 mentions) Anti pkcδ, by Cell Signaling Technology (2 mentions) Anti vimentin, by Cell Signaling Technology (2 mentions) Anti vinculin, by Merck Group (2 mentions) Anti β actin, by BD (2 mentions) Anti mypt1, by Cell Signaling Technology (2 mentions) Anti β actin clone ac 74, by Merck Group (1 mentions) Rho assay reagent, by Merck Group (1 mentions) Alliance mini wl2m system, by Uvitec (1 mentions) Anti pdcd10, by Merck Group (1 mentions) Anti cyclin d1 sc 8396, by Santa Cruz Biotechnology (1 mentions) Anti parp, by Cell Signaling Technology (1 mentions) Blebbistatin, by Abcam (1 mentions)

Close spelling variants of this product

Anti-phospho MYPT1 (T850) Anti-phospho-MYPT1 (Thr696) antibody Phospho-MYPT1 Anti-MYPT1 MYPT1

5 protocols using anti phospho mypt1

1

Western Blot Analysis of Cellular Signaling

Cited 1 time
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Whole cell extracts were prepared and transferred to membranes with standard methods as described before [15] (link). Membranes were incubated overnight at 4°C with the following antibodies: anti-PDCD10 (#SAB1305161), anti-β-Actin (clone AC-74, #A2228) from Sigma-Aldrich; anti-phospho cofilin (sc-12912), anti-cofilin (sc-33779) and anti- Cyclin D1 (sc-8396) from Santa Cruz (Dallas, TX, USA); anti-phospho-Mypt1 (#5163), anti-Mypt1 (#2634), anti-phospho-MLC2 (#3671), anti-MLC2 (#8505) and anti-PARP (#9542) from Cell Signaling Technology (Danvers, MA, USA); anti-cleaved Caspase 3 (2305-PC-100) from Trevigen (Gaithersburg, MD, USA). GTP-bound Rho was assayed with Rho assay reagent according to manifacturer instructions (#17-294; Millipore, Burlington, MA, USA). Chemio-luminescence was detected with Alliance Mini WL2M system (Uvitec, Cambridge, UK).
De Marco C., Zoppoli P., Rinaldo N., Morganella S., Morello M., Zuccalà V., Carriero M.V., Malanga D., Chirillo R., Bruni P., Malzoni C., Di Vizio D., Venturella R., Zullo F., Rizzuto A., Ceccarelli M., Ciliberto G, & Viglietto G. (2021). Genome-wide analysis of copy number alterations led to the characterisation of PDCD10 as oncogene in ovarian cancer. Translational Oncology, 14(3), 101013.
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2

Western Blot Analysis of Key Signaling Proteins

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Western blots were carried out as previously described (35 (link)), using the following antibodies: anti-Rac1 clone 23A8 (catalog # 05-389, Upstate Biotechnology, Lake Placid, NY), anti-phospho-Erk1/2 (catalog # 9101S), anti-PKCδ (catalog # 2058S), anti-PKCε (catalog # 2683S), anti-phosphoSmad2/3 (catalog # 8828S), anti-phospho-MYPT1 (catalog # 5163S), anti-vimentin (catalog # 3390S), anti-Cdc42 (catalog # 2466S), anti-RhoA (catalog # 2117S, Cell Signaling Technology, Beverly, MA), anti-PKCε (catalog # sc-208, Santa Cruz Biotechnology, Dallas, TX), anti-vinculin (catalog # V9131, Sigma-Aldrich, St. Louis, MO), anti-β-actin (catalog # A5441, BD Biosciences, Franklin Lakes, NJ), and anti E-cadherin (catalog # MAB1838, RD Systems, Minneapolis, MN).
Casado-Medrano V., Barrio-Real L., Wang A., Cooke M., Lopez-Haber C, & Kazanietz M.G. (2019). DISTINCTIVE REQUIREMENT OF PKCε IN THE CONTROL OF Rho GTPases IN EPITHELIAL AND MESENCHYMALLY-TRANSFORMED LUNG CANCER CELLS. Oncogene, 38(27), 5396-5412.
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3

Molecular Markers of Tight Junction Regulation

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Primary antibodies for Western blot include anti–NKA β1 subunit (Upstate, 05-382), anti-occludin (Invitrogen, 71-1500), anti–zo-1 (Invitrogen, 61-7300), anti–zo-2 (Invitrogen, 71-1400), anti-actin (MilliporeSigma, A2066), anti-GAPDH (MilliporeSigma, CB1-001), anti–NKA β2 subunit (Abcam, ab185210), anti-DDK (OriGene, TA50011-100), anti-MRCKα (Santa Cruz Biotechnology Inc., sc-374568), anti-MYPT1 (Cell Signaling Technology, 2634S), anti–phospho-MYPT1 (Thr696, Cell Signaling Technology, 5163S), anti-MLC2 (Cell Signaling Technology, 3672S), and anti–phospho-MLC2 (Ser19, Cell Signaling Technology, 3671S). The primary antibodies for immunofluorescence include anti–occludin Alexa Fluor594 (Invitrogen, 331594), anti–zo-1-Alexa Fluor594 (Invitrogen, 339194), and anti-MRCKα (Thermo Fisher Scientific, PA1-10038). The inhibitor for MRCKα BDP5290 was purchased from Aobious. Myosin inhibitor blebbistatin was purchased from Abcam.
Bai H., Zhou R., Barravecchia M., Norman R., Friedman A., Yu D., Lin X., Young J.L, & Dean D.A. (2021). The Na+, K+-ATPase β1 subunit regulates epithelial tight junctions via MRCKα. JCI Insight, 6(4), e134881.
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4

Immunoblot Analysis of Hepatocytes

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For immunoblot analysis 50μg of isolated hepatocytes lysate was resolved by a 4–20% gradient gel, transferred to a nitrocellulose membrane, and blotted with the appropriate primary antibodies. Membranes were incubated with peroxidase-conjugated secondary antibody (Cell signaling, Danvers, MA, USA). Protein bands were visualized using an enhanced chemiluminescence reagent and digitized using a CCD camera (ChemiDoc®, BioRad, Hercules, CA, USA). A rabbit anti-cleaved caspase 3, anti-caspase 3, anti-phospho-MYPT1, or anti-MYPT1 was purchased from Cell Signaling and anti-beta actin was purchased from GeneTex (Irvine, CA, USA). Protein load was verified using GAPDH (GeneTex), or PORIN (GeneTex) antibody.
Eguchi A., Lazaro R.G., Wang J., Kim J., Povero D., Willliams B., Ho S.B., Stärkel P., Schnabl B., Ohno-Machado L., Tsukamoto H, & Feldstein A.E. (2016). Extracellular vesicles released by hepatocytes from gastric infusion model of ALD contain a miRNA barcode that can be detected in blood. Hepatology (Baltimore, Md.), 65(2), 475-490.
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5

Western Blot Analysis of Key Signaling Proteins

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Western blots were carried out as previously described (35 (link)), using the following antibodies: anti-Rac1 clone 23A8 (catalog # 05-389, Upstate Biotechnology, Lake Placid, NY), anti-phospho-Erk1/2 (catalog # 9101S), anti-PKCδ (catalog # 2058S), anti-PKCε (catalog # 2683S), anti-phosphoSmad2/3 (catalog # 8828S), anti-phospho-MYPT1 (catalog # 5163S), anti-vimentin (catalog # 3390S), anti-Cdc42 (catalog # 2466S), anti-RhoA (catalog # 2117S, Cell Signaling Technology, Beverly, MA), anti-PKCε (catalog # sc-208, Santa Cruz Biotechnology, Dallas, TX), anti-vinculin (catalog # V9131, Sigma-Aldrich, St. Louis, MO), anti-β-actin (catalog # A5441, BD Biosciences, Franklin Lakes, NJ), and anti E-cadherin (catalog # MAB1838, RD Systems, Minneapolis, MN).
Casado-Medrano V., Barrio-Real L., Wang A., Cooke M., Lopez-Haber C, & Kazanietz M.G. (2019). DISTINCTIVE REQUIREMENT OF PKCε IN THE CONTROL OF Rho GTPases IN EPITHELIAL AND MESENCHYMALLY-TRANSFORMED LUNG CANCER CELLS. Oncogene, 38(27), 5396-5412.
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