Phospho yaps127

Manufactured by Cell Signaling Technology

Sourced in United States

Phospho YAPS127 is a lab equipment product that enables the detection and quantification of phosphorylated YAP (Serine 127) protein. It is a tool used in biological research to study cell signaling pathways.

Automatically generated - may contain errors

Lab products found in correlation

Lats1, by Cell Signaling Technology (3 mentions) Gapdh, by Merck Group (3 mentions) Mcl 1, by Santa Cruz Biotechnology (2 mentions) Phospho yapy357, by Abcam (2 mentions) Total yap, by Santa Cruz Biotechnology (2 mentions) Phospho src y416, by Cell Signaling Technology (2 mentions) Prolong antifade with 4 6 diamidino 2 phenylindole dapi, by Thermo Fisher Scientific (2 mentions) Anti flag m2, by Merck Group (2 mentions) Phospho nf κb, by Cell Signaling Technology (2 mentions) Anti vimentin, by Agilent Technologies (2 mentions) Phospho iκbα, by Cell Signaling Technology (2 mentions) Stat3, by Cell Signaling Technology (2 mentions) Phospho stat3, by Cell Signaling Technology (2 mentions) E cadherin, by Cell Signaling Technology (2 mentions) Nf κb, by Cell Signaling Technology (2 mentions) β catenin, by BD (2 mentions) Bay 60 6583, by Bio-Techne (1 mentions) Male sprague dawley rat, by Charles River Laboratories (1 mentions) Culture media and additives, by Thermo Fisher Scientific (1 mentions) Anti brdu antibody, by Merck Group (1 mentions)

12 protocols using phospho yaps127

Sprague Dawley Rat Biochemical Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Male Sprague Dawley (SD) rats were obtained from Charles River. Culture media and additives were obtained from Invitrogen. All chemicals were obtained from Sigma unless otherwise stated. BAY60-6583 was from Tocris. Antibodies to YAP, phospho-YAPS127, phospho-YAPS397, TAZ, pan-TEAD and phospho-Retinoblastoma protein were from Cell Signalling Technologies. Anti-BrDU antibody was from Sigma.

Kimura T.E., Duggirala A., Smith M.C., White S., Sala-Newby G.B., Newby A.C, & Bond M. (2016). The Hippo pathway mediates inhibition of vascular smooth muscle cell proliferation by cAMP. Journal of Molecular and Cellular Cardiology, 90, 1-10.

+ Open protocol

+ Expand

Investigating YAP Signaling Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Crenolanib (Selleck Chemicals) and Dasatinib (TSZ Chemicals) was added to cells at final concentrations from 0.1–10μM. The following primary antisera were used for immunoblot analysis: PDGF Receptor (28E1 Cell Signaling, Danvers, MA), actin (C-11 Santa Cruz, Dallas, TX), phospho YAP^Y357 (ab62751 abcam, Cambridge, MA), phospho YAP^S127 (4911 Cell Signaling), total YAP (63.7 Santa Cruz), lamin B1 (D8P3U Cell Signaling), GAPDH (MAB374 Millipore, Temecula, CA), phospho Src^Y416 (2101 Cell Signaling), Src (L4A1 Cell Signaling), and Mcl-1 (S-19 Santa Cruz). The following primary monoclonal antibodies were used for immunofluorescence: total YAP (63.7 Santa Cruz) and anti-FLAG M2 (F1804 Sigma Adrich). For ChIP immunoprecipitation assays we employed an anti-YAP monoclonal antibody (63.7 Santa Cruz). ProLong Antifade with 4′,6-diamidino-2-phenylindole (DAPI, Life Technologies) was used for nuclear staining. The following primers used are listed in Table 1.

Smoot R.L., Werneburg N.W., Sugihara T., Hernandez M.C., Yang L., Mehner C., Graham R.P., Bronk S.F., Truty M.J, & Gores G.J. (2017). Platelet-Derived Growth Factor Regulates YAP Transcriptional Activity via Src Family Kinase Dependent Tyrosine Phosphorylation. Journal of cellular biochemistry, 119(1), 824-836.

+ Open protocol

+ Expand

Antibody-based analysis of Hippo pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against YAP (#4912), YAP/TAZ (#8418), phospho-YAP S127 (#4911), LATS1 (#3477, 1:200), phospho-LATS1 Thr1079 (#9159, 1:200), MOB1 (#3863), phosphor-MOB1T35 (#8699), MST1 (#3682) were purchased from Cell Signaling, USA, anti-14:3:3 ε (sc-393177), TAZ (sc-48805), Ubiquitin (sc-8017) and Lamin B1 (sc-377001) were from SantaCruz Biotechnology. Anti-LATS1 ab70562 (abcam) and anti-MOB1A/B sc-161867 (Santa Cruz) were used in the mouse cell line with 1:500 dilution. Anti-Hsp27 (1:5000) was from ENZO lifesciences. MG132 was from Millipore (#474791).

Vahid S., Thaper D., Gibson K.F., Bishop J.L, & Zoubeidi A. (2016). Molecular chaperone Hsp27 regulates the Hippo tumor suppressor pathway in cancer. Scientific Reports, 6, 31842.

+ Open protocol

+ Expand

Western Blot Analysis of Kidney Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot analysis was performed on stage E13.5 Nf2^UB−/− kidneys or cultured Yap^UB-OE kidneys following doxycycline induction. Unless stated otherwise each western blot lane represents one animal (two kidneys). Kidneys were mechanically homogenized and lysed in RIPA buffer supplemented with proteasome and phosphatase inhibitors. Western blot analysis was performed following standard protocols with the following primary antibodies: CALBINDIN (C9848, Sigma, 1:1,000), BAF155 (SC-9748, SantaCruz Biotechnology, 1:2,000), GAPDH (R9545, Sigma, 1:7,500), HA (11867423001, Roche, 1:1,000), LATS1 (#3477, Cell Signaling Technology, 1:1,000), MST1 (#3682, Cell Signaling Technology, 1:2,000), NF2 (HPA003097, Sigma Prestige Antibodies, 1:2,000), phosho-LATS1 S909 (#9157, Cell Signaling Technology, 1:500), phospho-MST1/2 T183/T180 (#3681, Cell Signaling Technology, 1:2,000), phospho-YAP S127 (#4911, Cell Signaling Technology, 1:2,000) and YAP (sc-101199, SantaCruz Biotechnology, 1:2,000). All uncropped western blots can be found in Supplementary Fig. 8.

Reginensi A., Enderle L., Gregorieff A., Johnson R.L., Wrana J.L, & McNeill H. (2016). A critical role for NF2 and the Hippo pathway in branching morphogenesis. Nature Communications, 7, 12309.

+ Open protocol

+ Expand

Density-Dependent Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown at either high or low density in culture and then lysed. Proteins extracted from the lysates were separated by SDS-PAGE and immunoblotted with anti-vimentin (Dako, Carpinteria, CA), beta-catenin (BD Biosciences, San Jose, CA), actin (Millipore, Billerica, MA), E-Cadherin, Stat3, phospho-Stat3, NFκB, phospho-NFκB, IκBα, phospho-IκBα, phospho-YAP (S127) (Cell Signaling, Boston, MA), and YAP (Santa Cruz, Dallas, TX).

Sharif G.M., Schmidt M.O., Yi C., Hu Z., Haddad B.R., Glasgow E., Riegel A.T, & Wellstein A. (2015). Cell growth density modulates cancer cell vascular invasion via Hippo pathway activity and CXCR2 signaling. Oncogene, 34(48), 5879-5889.

+ Open protocol

+ Expand

Western Blot Analysis of Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in ice-cold radioimmunoprecipitation assay (RIPA) buffer supplemented with a protease and phosphatase inhibitor cocktail (Roche). Whole-cell extracts were incubated with the indicated antibodies and protein A/G beads (Santa Cruz Biotechnology) overnight. The beads were washed with RIPA buffer three times and then boiled in SDS loading buffer. For Western blotting, cell extracts were separated by SDS-PAGE. After Western transfer, membranes were probed with antibodies against DDK (Origene ta50011), GFP (Santa Cruz biotechnology sc-101536), Glut3 (Santa Cruz biotechnology sc-74399), HA (Santa Cruz biotechnology sc-393579), PKM2 (Cell Signaling #3198), phospho-PKM2(Y705) (Cell Signaling #3827), YAP (Cell Signaling #12395), phospho-YAP(S127) (Cell Signaling #13008), actin (GeneTex GTX11003) and lamin C (GeneTex GTX101127). Bands were detected with an enhanced chemiluminescence system (Millipore). Western blotting was performed at least three times, and representative experiments are shown. Quantification was carried out using ImageJ software.

Kuo C.C., Ling H.H., Chiang M.C., Chung C.H., Lee W.Y., Chu C.Y., Wu Y.C., Chen C.H., Lai Y.W., Tsai I.L., Cheng C.H, & Lin C.W. (2019). Metastatic Colorectal Cancer Rewrites Metabolic Program Through a Glut3-YAP-dependent Signaling Circuit. Theranostics, 9(9), 2526-2540.

+ Open protocol

+ Expand

Comprehensive Cell Signaling Antibody Panel

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against phospho-S6K(T389) (#9205), S6K (#9202), phospho-4E-BP1(T37/46) (#9459), 4E-BP1 (#9452), phospho-Akt(S473) (#9271), phospho-Akt(T308) (#9275), Akt (#9272), phospho-TSC2(T1462) (#3611), phospho-TSC2(S1387) (#5584), phospho-TSC2(S1254) (#3616), TSC2 (#4308), mTOR (#2983), phospho-GSK3β(Ser9) (#5558), phospho-p38(T180/Y182) (#9216), p38 (#9212), phospho-MAPKAPK2(T222) (#3316), phospho-ERK1/2(T202/Y204) (#4370), ERK1/2 (#4695), phospho-AMPKβ1(S108) (#4181), phospho-p90RSK(S380) (#9341), phospho-Erk5(T218/Y220) (#3371), phospho-CaMKII(T286) (#3361), phospho-MST1(T183)/MST2(T180) (#3681), and phospho-YAP(S127) (#4911) proteins were purchased from Cell Signaling Technology. An antibody against phospho-TSC2(S664) was purchased from Abcam (ab133465). The mouse LAMP2 (ABL-93) antibody was obtained from Developmental Studies Hybridoma Bank. A monoclonal antibody recognizing human and mouse α-tubulin (#T9026) and an anti-FLAG (M2) (#F1804) antibody were purchased from Sigma. All antibodies were used in 1:1000 dilution for western blotting, except for the phospho-ERK, total ERK, TSC2, and p38 antibodies that were used in 1:2000.

Plescher M., Teleman A.A, & Demetriades C. (2015). TSC2 mediates hyperosmotic stress-induced inactivation of mTORC1. Scientific Reports, 5, 13828.

+ Open protocol

+ Expand

Investigating YAP Signaling Pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

Dasatinib (Selleckchem), KW-2449 (Selleckchem), sodium orthovanadate (Sigma), and/or latrunculin A (Abcam) was added to cells at final concentrations from 0.1–10 µM per experimental design. The following primary antisera were used for immunoblot analysis: actin (C-11 Santa Cruz, Dallas, TX), phospho YAP^Y357 (ab62751 abcam, Cambridge, MA), phospho YAP^S127 (4911 Cell Signaling), total YAP (63.7 Santa Cruz, 4912 CST), lamin B1 (D8P3U Cell Signaling), GAPDH (MAB374 Millipore, Temecula, CA), phospho SRC^Y416 (2101 Cell Signaling), SRC (L4A1 Cell Signaling), YES (3201 Cell Signaling), LCK (2657 Cell Signaling), FYN (4023 Cell Signaling), LYN (44 Santa Cruz), LATS1 (C66B Cell Signaling), phospho LATS1 (S909 Cell Signaling) and MCL-1 (S-19 Santa Cruz). The following primary monoclonal antibody were used for immunofluorescence and/or immunohistochemistry: total YAP (63.7 Santa Cruz), phospho YAP^Y357 (ab62751 abcam, Cambridge, MA), SOX9 (AB5535 Millipore) and anti-FLAG M2 (F1804 Sigma Adrich). ProLong Antifade with 4’,6-diamidino-2-phenylindole (DAPI, Life Technologies) was used for nuclear staining.

Sugihara T., Werneburg N.W., Hernandez M.C., Yang L., Kabashima A., Hirsova P., Yohanathan L., Sosa C., Truty M.J., Vasmatzis G., Gores G.J, & Smoot R.L. (2018). YAP Tyrosine Phosphorylation and Nuclear Localization in Cholangiocarcinoma Cells is Regulated by LCK and Independent of LATS Activity. Molecular cancer research : MCR, 16(10), 1556-1567.

+ Open protocol

+ Expand

Antibody Sources for Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies against IL-6 were obtained from Ab Cam (Cat# 191194), CD11B were acquired from Novus Biologicals (Cat# NB110-89474), and RALGDS from Abiocode (Cat# R1857-vp). Antibodies for Phospho-ERK (Cat# 9101), ERK (Cat# 9102), Phospho-AKT-Ser473 (Cat# 9271), Phospho-YAP-S127 (Cat# 4911), YAP (Cat# 4912), and Mouse and Rabbit secondary antibodies (Cat# 7076 and 7074) were obtained from Cell Signaling Technologies (Boston, MA). Epitope tag antibodies detecting GFP were purchased from Santa Cruz (Cat# SC-9996) and HA from Covance (Cat# MMS-101P). RASSF1A rabbit antibody was generated by Prosci (Poway CA) against the peptide “ELRELAPAGRAGKGRTRLER”.

Schmidt M.L., Hobbing K.R., Donninger H, & Clark G.J. (2018). RASSF1A deficiency enhances RAS driven lung tumorigenesis. Cancer research, 78(10), 2614-2623.

+ Open protocol

+ Expand

Immunofluorescence Analysis of YAP Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

EMT6 cells were seeded on glass coverslips in a 24-well plate and incubated O/N for attachment. After indicated treatments, samples were washed with 1X PBS and fixed with 4% PFA for 20 min. Then, Triton-X (0.5%) was used to permeabilize the cells for 20 min on a shaker. Samples were blocked with 3% BSA for 1 h on a shaker, washed three times with 1X PBS, and incubated with the following antibodies Phospho-YAP (S127) (Cell Signaling 13008), YAP (Cell Signaling 4912), or calreticulin (Cell Signaling D3E6) O/N on shaker at 4°C. The following day samples were washed and incubated for 1 h on a shaker with DyLight^™ 488 Donkey anti-rabbit IgG (minimal x-reactivity) antibody (BioLegend 406404). After incubation, samples were washed and incubated with Flash Phalloidin^™ Red 594 (BioLegend 424203) for 30 min on a shaker. Lastly, samples were washed and mounted in ProLong^™ Glass Antifade Mountant with NucBlue^™ Stain (Thermo Fisher P36981) and stored in the dark until imaging.

Hamsici S., Gunay G, & Acar H. (2022). Controllable membrane damage by tunable peptide aggregation with albumin. AIChE journal. American Institute of Chemical Engineers, 68(12), e17893.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!