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Pan tead

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Pan-TEAD is a recombinant protein that can be used as a tool to study the activity of TEAD transcription factors. TEAD proteins are key mediators of the Hippo signaling pathway and play a role in cell proliferation and growth control.

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Lab products found in correlation

Lats1, by Cell Signaling Technology (5 mentions) Yap taz, by Cell Signaling Technology (4 mentions) Myc tag, by Cell Signaling Technology (3 mentions) Gapdh, by Santa Cruz Biotechnology (3 mentions) Ha tag, by Cell Signaling Technology (2 mentions) Precast gradient gel, by Bio-Rad (2 mentions) α tubulin, by Merck Group (2 mentions) Lamin b1, by Cell Signaling Technology (2 mentions) Cyclophilin b, by Thermo Fisher Scientific (2 mentions) Histone h3, by Cell Signaling Technology (2 mentions) Ripa lysis buffer, by Merck Group (2 mentions) Flag tag (flag), by Merck Group (2 mentions) Gapdh, by Thermo Fisher Scientific (2 mentions) Hsp90, by BD (2 mentions) P mk2, by Cell Signaling Technology (2 mentions) Tead1, by Merck Group (2 mentions) Vinculin, by Merck Group (2 mentions) Flag tag (flag), by Abcam (2 mentions) Dykddddk tag, by Cell Signaling Technology (2 mentions) Lats2 a300 479a, by Fortis Life Sciences (2 mentions)

16 protocols using pan tead

1

Immunoprecipitation and Western Blot Assay

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Cultured HEK293T cells were lysed in lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5 mM EDTA, 1% Triton X-100, phosphatase inhibitor cocktail, cOmplete EDTA-free protease inhibitors cocktail) for 30 minutes at 4°C. The supernatants of the extracts were then used for the immunoprecipitation and western blot following the protocols described previously (Cotton et al., 2017 (link)). The primary antibodies used in these assays were: GAPDH (cat. 2118, 1:5000, Cell Signaling Technology, Danvers, MA, USA), YAP/TAZ (cat. 8418, 1:1000, Cell Signaling Technology), panTEAD (cat. 13295, 1:1000, Cell Signaling Technology), V5-tag (cat. 13202, 1:1000, Cell Signaling Technology), p300 (cat. 86377, 1:1000, Cell Signaling Technology), Flag-tag (cat. 2368, 1:1000, Cell Signaling Technology and cat. F9291, 1:1000, Sigma), HA-tag (cat. 3724 and cat. 2367, 1:1000, Cell Signaling Technology). HRP-conjugated secondary antibodies used for detection were obtained from Jackson Laboratories (Sacramento, CA, USA).
Guo S., Hu X., Cotton J.L., Ma L., Li Q., Cui J., Wang Y., Thakare R.P., Tao Z., Ip Y.T., Wu X., Wang J, & Mao J. (2024). VGLL2 and TEAD1 fusion proteins drive YAP/TAZ-independent transcription and tumorigenesis by engaging p300. bioRxiv.
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2

Sprague Dawley Rat Biochemical Assay

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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.
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3

PAX8 Immunoprecipitation and TEAD Analysis

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The nuclear fraction was isolated from KURAMOCHI cells (Cell Signaling Technology, Cell Fractionation Kit, 9038). Lysate (500 µg) was immunoprecipitated with 10 µg of a rabbit polyclonal antibody against PAX8 (Novus Biologicals, NBP1-32440) or equal amount of rabbit IgG using the Pierce Direct IP kit (Thermo Fisher Scientific, 26148) according to manufacturer’s protocol, followed by SDS-PAGE, transfer, and Western blot analysis using a rabbit monoclonal antibody to pan-TEAD (Cell Signaling Technology, 13295).
Elias K.M., Emori M.M., Westerling T., Long H., Budina-Kolomets A., Li F., MacDuffie E., Davis M.R., Holman A., Lawney B., Freedman M.L., Quackenbush J., Brown M, & Drapkin R. (2016). Epigenetic remodeling regulates transcriptional changes between ovarian cancer and benign precursors. JCI Insight, 1(13), e87988.
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4

Co-Immunoprecipitation of YAP, 14-3-3, and Huntingtin

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The co-immunoprecipitations studies were carried out as previously described69 (link). Briefly, using post-mortem human cortex homogenates, 150 μg of protein was incubated with either 5 μl anti-YAP antibody (Novus Biologicals), anti-14-3-3- antibody (ThermoFisher Scientific Waltham, MA), or anti-Htt MAB2166 antibody (Millipore) and GAL4 immunoprecipitation buffer (2.5 ml 5M NaCl, 500 μl EDTA, 500 μl P-40, 0.3 g Tris Base, and dH2O to 50 ml). After incubation at 4 °C for 3.5 hours, magnetic protein A beads (Invitrogen) were added (20 μl per sample) and then left spinning overnight at 4 °C. Samples were placed on a magnetic rack to facilitate removal of the supernatant, and washed with GAL4. Samples were then boiled with 20 μl of sample buffer for 5 min at 95°.
For the input samples, 75 μg of protein was resuspended in sample buffer and boiled at 95 °C for 5 min. The samples were then loaded into a 4–20% glycine gel and the protocol for a Western blot was followed. The following were used to immunoblot: pan-TEAD (Cell Signaling Technology), 14-3-3 (ThermoFisher Scientific), Huntingtin (Millipore), pYAP (Cell Signaling Technology).
Mueller K.A., Glajch K.E., Huizenga M.N., Wilson R.A., Granucci E.J., Dios A.M., Tousley A.R., Iuliano M., Weisman E., LaQuaglia M.J., DiFiglia M., Kegel-Gleason K., Vakili K, & Sadri-Vakili G. (2018). Hippo Signaling Pathway Dysregulation in Human Huntington’s Disease Brain and Neuronal Stem Cells. Scientific Reports, 8, 11355.
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5

Western Blot Analysis of Cellular Proteins

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Cells were lysed in RIPA lysis buffer (Millipore) containing protease inhibitors and phosphatase inhibitors (GenDEPOT). Proteins were resolved on 4–20% precast gradient gels (Bio-Rad) and transferred to a PVDF membrane. After blocking with 5% non-fat milk in Tris-buffered saline with 0.05% Tween-20 (TBST), membranes were incubated with the primary antibody followed by the secondary antibody conjugated with horseradish peroxidase. After washing, the bands were visualized with enhanced chemiluminescence substrate (Denville). Primary antibodies used are as follows: antibodies against pan-TEAD (1:1,000, Cell Signaling Technology, #13295), FLAG (1:5,000, Sigma, #F7425), HA (1:2,000, Santa Cruz Biotechnology, #sc-7392), cyclophilin B (1:5,000, ThermoFisher Scientific, #PA1–027A), YAP (1:1,000, Cell Signaling Technology, #14074), histone H3 (1:1,000, Cell Signaling Technology, #9715), Lamin B1 (1:1,000, Cell Signaling Technology, #12586), α-tubulin (1:1,000, Sigma, #T5168), HSP90 (1:5,000, BD Biosciences, #610419), and GAPDH (1:1,000, ThermoFisher Scientific, #MA5–15738).
Kim J., Piao H.L., Kim B.J., Yao F., Han Z., Wang Y., Xiao Z., Siverly A.N., Lawhon S.E., Ton B.N., Lee H., Zhou Z., Gan B., Nakagawa S., Ellis M.J., Liang H., Hung M.C., You M.J., Sun Y, & Ma L. (2018). Long non-coding RNA MALAT1 suppresses breast cancer metastasis. Nature genetics, 50(12), 1705-1715.
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6

Western Blot Analysis of Cellular Proteins

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Cells were lysed in RIPA lysis buffer (Millipore) containing protease inhibitors and phosphatase inhibitors (GenDEPOT). Proteins were resolved on 4–20% precast gradient gels (Bio-Rad) and transferred to a PVDF membrane. After blocking with 5% non-fat milk in Tris-buffered saline with 0.05% Tween-20 (TBST), membranes were incubated with the primary antibody followed by the secondary antibody conjugated with horseradish peroxidase. After washing, the bands were visualized with enhanced chemiluminescence substrate (Denville). Primary antibodies used are as follows: antibodies against pan-TEAD (1:1,000, Cell Signaling Technology, #13295), FLAG (1:5,000, Sigma, #F7425), HA (1:2,000, Santa Cruz Biotechnology, #sc-7392), cyclophilin B (1:5,000, ThermoFisher Scientific, #PA1–027A), YAP (1:1,000, Cell Signaling Technology, #14074), histone H3 (1:1,000, Cell Signaling Technology, #9715), Lamin B1 (1:1,000, Cell Signaling Technology, #12586), α-tubulin (1:1,000, Sigma, #T5168), HSP90 (1:5,000, BD Biosciences, #610419), and GAPDH (1:1,000, ThermoFisher Scientific, #MA5–15738).
Kim J., Piao H.L., Kim B.J., Yao F., Han Z., Wang Y., Xiao Z., Siverly A.N., Lawhon S.E., Ton B.N., Lee H., Zhou Z., Gan B., Nakagawa S., Ellis M.J., Liang H., Hung M.C., You M.J., Sun Y, & Ma L. (2018). Long non-coding RNA MALAT1 suppresses breast cancer metastasis. Nature genetics, 50(12), 1705-1715.
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7

Antibody Optimization for Western Blot, IHC, and IF

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The following antibodies were purchased from Cell Signaling and used at the indicated dilution for western blot analysis, immunohistochemistry, and immunofluorescence: pan-TEAD (13295, 1:1000), p38 MAPK (8690, 1:1000), phospho-p38 MAPK (4511, 1:1000), YAP (14074, 1:1000), TAZ (4883, 1:1000), Lats1 (3477, 1:1000), p-MK2 (3007, 1:1000), p-ERK (4370, 1:1000), DYKDDDDK tag (2368, 1:1000), Myc tag (2276, 1:1000), p38α (2371, 1:1000), p38β (2339, 1:1000), p38γ (2307, 1:1000), and p38δ (2308, 1:1000). The following antibodies were purchased from Santa Cruz Biotechnology and used at the indicated dilution for western blot analysis and immunofluorescence: YAP (sc-101199, 1:1000), HA (sc-7392, 1:5000), Myc (sc-40, 1:5000), GAPDH (sc-25778, 1:1000). TEAD4 (ab58310, 1:1000) was purchased from Abcam, Flag (A8592, 1:10,000) and vinculin (V9131, 1:5000) was purchased from Sigma, TEAD1 (610923, 1:1000) was purchased from BD Biosciences, Lats2 (A300-479A, 1:1000) was purchased from Bethyl Laboratories, and NFAT5 (bs-9473R OWL 1:1000) was purchased from One World Lab.
Lin K.C., Moroishi T., Meng Z., Jeong H.S., Plouffe S.W., Sekido Y., Han J., Park H.W, & Guan K.L. (2017). Regulation of Hippo pathway transcription factor TEAD by p38 MAPK-induced cytoplasmic translocation. Nature cell biology, 19(8), 996-1002.
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8

Immunoblotting and Immunohistochemistry Antibodies

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Antibodies for immunohistochemistry: NONO (sc‐166702, Santa Cruz Biotechnology) was used at a 1/200 dilution and secondary antibodies at a 1/200 dilution. Antibodies for immunoblotting: µ‐Actin (#3700, dilution at 1/20000), TAZ (#4883S, dilution at 1/1000), Pan‐Tead (#13 295, dilution at 1/2000), Rpb1 (#2629, dilution at 1/3000), p‐YAPS127 (#13 008, dilution at 1/1000), YAP (#12 395, dilution at 1/2000), p‐TAZS89 (#59 971, dilution at 1/1000), Myc (#2272, dilution at 1/3000), p‐Lats1T1079 (#8654, dilution at 1/500), and Lats1 (#3477, dilution at 1/1000), all from Cell Signaling Technologies; HA (MMS‐101P, Biolegend, dilution at 1/3000); and goat anti‐rabbit HRP‐conjugated antibody (#7074S, dilution at 1/5000) and goat anti‐mouse HRP‐conjugated antibody, (#7076S, dilution at 1/5000) both from Cell Signaling Technologies.
Wei Y., Luo H., Yee P.P., Zhang L., Liu Z., Zheng H., Zhang L., Anderson B., Tang M., Huang S, & Li W. (2021). Paraspeckle Protein NONO Promotes TAZ Phase Separation in the Nucleus to Drive the Oncogenic Transcriptional Program. Advanced Science, 8(24), 2102653.
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9

Immunoblotting Analysis of Hippo Pathway

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Cells or tissues were lysed in RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl at pH 7.4, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM PMSF) with protease inhibitors (Roche). The proteins were separated on SDS–polyacrylamide gels and transferred onto PVDF membranes (Millipore). The blots were probed with antibodies against YAP (Cell Signaling, #4912), phospho-YAP (Ser112; Cell Signaling, #4911), phospho-YAP (Ser366; Cell Signaling, #13619), Lats1 (Cell Signaling, #3477), Lats2 (Cell Signaling, #13646), Mst1 (Cell Signaling, #ab51134), NF2 (Sigma, HPA003097), Mst2 (Epitomics, 1943-1), CTGF (Santa Cruz Biotechnology, sc-14939), α-tubulin (Developmental Studies Hybridoma Bank, 12G10), PAN-TEAD (Cell Signaling, #13295), MOB1 (Cell Signaling, #3863), phospho-MOB1 (Cell Signaling, #8699), YAP, and TAZ (Cell Signaling, #8418) and normalized by Actin (Millipore, MAB1501). Signals were detected and quantified by a LI-COR infrared imaging system. The antibodies used for coimmunoprecipitation were 14-3-3 (Santa Cruz Biotechnology, sc-732) and YAP (Novus Biologicals, NB110-58358).
Chen Q., Zhang N., Xie R., Wang W., Cai J., Choi K.S., David K.K., Huang B., Yabuta N., Nojima H., Anders R.A, & Pan D. (2015). Homeostatic control of Hippo signaling activity revealed by an endogenous activating mutation in YAP. Genes & Development, 29(12), 1285-1297.
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10

Antibody Optimization for Western Blot, IHC, and IF

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The following antibodies were purchased from Cell Signaling and used at the indicated dilution for western blot analysis, immunohistochemistry, and immunofluorescence: pan-TEAD (13295, 1:1000), p38 MAPK (8690, 1:1000), phospho-p38 MAPK (4511, 1:1000), YAP (14074, 1:1000), TAZ (4883, 1:1000), Lats1 (3477, 1:1000), p-MK2 (3007, 1:1000), p-ERK (4370, 1:1000), DYKDDDDK tag (2368, 1:1000), Myc tag (2276, 1:1000), p38α (2371, 1:1000), p38β (2339, 1:1000), p38γ (2307, 1:1000), and p38δ (2308, 1:1000). The following antibodies were purchased from Santa Cruz Biotechnology and used at the indicated dilution for western blot analysis and immunofluorescence: YAP (sc-101199, 1:1000), HA (sc-7392, 1:5000), Myc (sc-40, 1:5000), GAPDH (sc-25778, 1:1000). TEAD4 (ab58310, 1:1000) was purchased from Abcam, Flag (A8592, 1:10,000) and vinculin (V9131, 1:5000) was purchased from Sigma, TEAD1 (610923, 1:1000) was purchased from BD Biosciences, Lats2 (A300-479A, 1:1000) was purchased from Bethyl Laboratories, and NFAT5 (bs-9473R OWL 1:1000) was purchased from One World Lab.
Lin K.C., Moroishi T., Meng Z., Jeong H.S., Plouffe S.W., Sekido Y., Han J., Park H.W, & Guan K.L. (2017). Regulation of Hippo pathway transcription factor TEAD by p38 MAPK-induced cytoplasmic translocation. Nature cell biology, 19(8), 996-1002.
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