Paxillin py118

Manufactured by Cell Signaling Technology

Paxillin pY118 is a phosphorylation-specific antibody that recognizes the tyrosine phosphorylation of paxillin at position 118. Paxillin is a focal adhesion-associated protein that plays a role in cell adhesion and signaling.

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Paxillin (34 citations) Anti-pY118-paxillin (2 citations)

3 protocols using paxillin py118

Western Blot Analysis of Cellular Proteins

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Cell lysates were prepared with RIPA. Protein in the supernatant was extracted, and its concentration was measured using the BCA Protein Assay Kit. An equal amount (50 ug) of total cellular protein was electrophoresed by denaturing 12.5% SDS-PAGE and transferred to 0.22um polyvinylidene difluoride (PVDF) membranes (Millipore, MA, USA). The locations of proteins of interest were detected by primary antibodies for overnight at 4°C. ERp19 antibody was from Abcam;GAPDH antibody was from Kangchen Bio-tech; FAK, FAK pY397, paxillin, paxillin pY118, ERK1/2 and p-ERK1/2 antibodies were from Cell Signaling Biotechnology. After HRP conjugated-secondary antibody bound to the primary antibodies, the proteins were visualized using enhanced chemiluminescence (ECL) reagent.

Wu J., Chen X.H., Wang X.Q., Yu Y., Ren J.M., Xiao Y., Zhou T., Li P, & Xu C.D. (2015). ERp19 contributes to tumorigenicity in human gastric cancer by promoting cell growth, migration and invasion. Oncotarget, 6(14), 11794-11805.

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Focal Adhesion Protein Analysis

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Antibodies used include: mouse monoclonal antibody (mAb) vinculin (MAB674; Millipore), mouse mAb talin (8d4; Sigma), rat mAbβ₁-integrin (AIIBII), rabbit mAb paxillin (Y113; Abcam), rabbit mAb FAK pY397 (141-9; Invitrogen), rabbit polyclonal antibody (pAb) α₅-integrin (AB1928; Millipore), mouse mAb MUC1 (HMPV; BD Pharminigen), hamster mAb MUC1 (CT2; Thermo Scientific), rabbit mAb Src Family pY416 (D49G4; Cell Signaling), mouse mAb FAK (77; BD Transduction Laboratories), rabbit pAb paxillin pY118 (2541; Cell Signaling), rabbit mAb pan-AKT (C67E7; Cell Signaling), rabbit pAb AKT pS473 (9271; Cell Signaling); rabbit mAb ERK1/2 pT202/pT204 (197G2; Cell Signaling); rabbit pAb ERK1/2 (9102; Cell Signaling); rabbit mAb Gapdh (14C10; Cell Signaling); Alexa 488 and Alexa 568 conjugated goat anti-mouse and anti-rabbit mAbs (Invitrogen); FITC conjugated anti-hamster mAbs; Cy5-conjugated goat anti-mouse and rabbit mAbs (Jackson); and HRP conjugated anti-rabbit and anti-mouse mAbs. Chemical inhibitors used in these studies include ROCK inhibitor Y-27632 (Cayman Chemical), myosin-II inhibitor (−)-blebbistatin (Cayman Chemical), FAK inhibitor FAK inhibitor 14 (Tocris), MEK inhibitor U0126 (Cell Signaling), PI(3)K inhibitor Wortmannin (Cell Signaling), Src inhibitor Src I1 (Sigma), and DiI (Molecular Probes).

Paszek M.J., DuFort C.C., Rossier O., Bainer R., Mouw J.K., Godula K., Hudak J.E., Lakins J.N., Wijekoon A.C., Cassereau L., Rubashkin M.G., Magbanua M.J., Thorn K.S., Davidson M.W., Rugo H.S., Park J.W., Hammer D.A., Giannone G., Bertozzi C.R, & Weaver V.M. (2014). The cancer glycocalyx mechanically primes integrin-mediated growth and survival. Nature, 511(7509), 319-325.

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Western Blot Antibody Library

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Antibodies for western blot detection included actin (I-19, Santa Cruz Biotechnology, cat. n. sc-1616, RRID: AB_630836), Akt (Cell Signaling Technology, cat. n. 9272, RRID: AB_329827), Akt pS473 (Cell Signaling Technology, cat. n. 9271, RRID: AB_329825), Cas (BD Transduction Laboratories, cat. n. 610271, RRID: AB_397666), Cas pY410 (Cell Signaling Technology, cat. n. 4011, RRID: AB_2274823), Erk1/2 (Promega), phospho-Erk1/2 (Promega, cat. n. V1141, RRID: AB_430839), FAK (C-20, Santa Cruz Biotechnology, cat. n. sc-558, RRID: AB_2300502), FAK pY397 (Invitrogen, cat. n. 44–624G, RRID: AB_2533701), FAK pY861 (Abgent, cat. n. AJ1285f, RRID: AB_10818227), paxillin (BD Transduction Laboratories, cat. n. 610052, RRID: AB_397464), paxillin pY118 (Cell Signaling Technology, cat. n. 2541, RRID: AB_2174466), Src (clone 184Q20, Invitrogen, cat. n. AHO1152, RRID: AB_2536324), Src pY418 (Cell Signaling Technology, cat. n. 2101, RRID: AB_331697), Stat3 (C-20, Santa Cruz Biotechnology, cat. n. sc-482, RRID: AB_632440), and Stat3 pY705 (Cell Signaling Technology, cat. n. 9131, RRID: AB_331568).

Koudelková L., Pelantová M., Brůhová Z., Sztacho M., Pavlík V., Pánek D., Gemperle J., Talacko P., Brábek J, & Rösel D. (2023). Phosphorylation of tyrosine 90 in SH3 domain is a new regulatory switch controlling Src kinase. eLife, 12, e82428.

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