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P lats1t1079

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P‐Lats1T1079 is a phospho-specific antibody that detects Lats1 protein phosphorylated at threonine 1079. This antibody is used for the detection and analysis of Lats1 phosphorylation in cell and tissue samples.

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Lab products found in correlation

Lats1, by Cell Signaling Technology (4 mentions) Pyaps127, by Cell Signaling Technology (3 mentions) P tazs89, by Cell Signaling Technology (1 mentions) Hrp conjugated goat anti rabbit antibody, by Cell Signaling Technology (1 mentions) Hrp conjugated goat anti mouse antibody, by Cell Signaling Technology (1 mentions) Pan tead, by Cell Signaling Technology (1 mentions) Anti flag tag, by CWBIO (1 mentions) Gapdh, by ZSGB-BIO (1 mentions) Chemiluminescence reagent, by Merck Group (1 mentions) Gadph, by Proteintech (1 mentions) Lamin a c, by Proteintech (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Hrp conjugated secondary antibody, by Proteintech (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) P yap, by Cell Signaling Technology (1 mentions) P110 pt1061 polyclonal antibody, by Cell Signaling Technology (1 mentions) Bis tris precast gel, by Thermo Fisher Scientific (1 mentions) Pcreb s133, by Cell Signaling Technology (1 mentions) P mob1 t35, by Cell Signaling Technology (1 mentions) Dc protein assay, by Bio-Rad (1 mentions)

4 protocols using p lats1t1079

1

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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2

Western Blot Analysis of Signaling Proteins

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Western blotting analysis performed as previously described [28 (link), 29 (link)]. The following antibodies were used: S100A8 (1/1000; R&D, MAB4570); S100A9 (1/1000; Santa-Cruz,8114); MST1 (1/1000; Cell Signaling Technology, 3682, Boston, USA); LATS1 (1/1000; Cell Signaling Technology, 3477); pLATS1-T1079 (1/1000; Cell Signaling Technology, 8654); YAP (1/1000; Cell Signaling Technology, 4912); pYAP-S127 (1/1000; Cell Signaling Technology, 13008S); anti-Flag tag (CWBIO, CW0287A, Beijing, China); anti-His tag (MBL, D291–3, NAGOYA, JAPAN). GAPDH (ZSGB-BIO, TA-08, Beijing, China) was used as loading control.
Li Y., Kong F., Jin C., Hu E., Shao Q., Liu J., He D, & Xiao X. (2019). The expression of S100A8/S100A9 is inducible and regulated by the Hippo/YAP pathway in squamous cell carcinomas. BMC Cancer, 19, 597.
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3

Western Blotting Protein Detection

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Proteins extract were obtained from RIPA lysis buffer [EDTA (1 mM), NaCl (150 mM), Tris-HCl (50 mM, pH = 7.4), sodium deoxycholate (1%), Triton X-100(1%) and SDS (0.1%)] containing phosphatase inhibitors and protease inhibitor (Roche, NJ, USA) and then were centrifuged 20 min (12,000 rpm, 4 °C). The protein lysates were electrophoresed using SDS-PAGE and transferred to polyvinyl difluoride (PVDF) (Bio-Rad, CA, USA). Membranes were blocked with 5% nonfat milk for two hours and incubated with the primary antibody in a certain environment (4 °C, overnight). The second day, membranes were cleaned three times (10 min each time) by TBST and incubated in HRP-conjugated secondary antibody (Proteintech) (one hour, room temperature). The membranes were photographed by the chemiluminescence reagent (Millipore). LaminA/C and GADPH act as controls. Nuclear and Cytoplasmic Extraction Reagents (Pioneer Biotechnology, Xi’an, China) was used for cell fractionation assays. Mst2, p-Lats1T1079, Lats1, p-YAP S127, YAP, E-cadherin and ZO-1 antibodies were obtained from CST (Cell Signaling Technology, USA.) The LaminA/C, GADPH, Vimentin primary antibodies were obtained from Proteintech, China. The CRABP1, CRABP2 and ER primary antibodies were obtained from Abcam, UK. The Flag primary antibodies were obtained from Sigma.
Feng X., Zhang M., Wang B., Zhou C., Mu Y., Li J., Liu X., Wang Y., Song Z, & Liu P. (2019). CRABP2 regulates invasion and metastasis of breast cancer through hippo pathway dependent on ER status. Journal of Experimental & Clinical Cancer Research : CR, 38, 361.
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4

Western Blot Analysis of AKT Pathway

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Cell lysates were prepared in the cell lysis buffer [50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 1 mM EDTA and 1% NP-40 with one tablet (per 10 mL) of protease and phosphatase inhibitor (Life Technologies)]. After incubation on ice for 30 min, lysates were centrifuged at top speed for 10 min and supernatant were collected and proteins were quantified using Protein DC assay (BioRad). Proteins were running and separated on 4%–12% Bis-Tris Pre-cast Gel (Life Technologies) using MOPS buffer and transferred to PVDF membrane at 350 mA for 1.5 h. Membranes were blocked in 5% non-fat milk in TBST and incubated with primary antibody overnight. Primary antibody against pAKT (S473) [#3787, #4060], pAKT (T308) [#4056], pAKT2 (S474) [#8599], pan AKT [#2920], AKT1 [#2967], AKT2 [#3063], pCREB (S133) [#9198], pLATS1 (T1079) [#8654], pMOB1 (T35) [#8699], pYAP (S127) [#13008], pPKA substrate motif antibody [#9624], p110a [#4249], CREB [#4820], MOB1 [#13730], LATS1 [#9153], MST1 [#14946] and GAPDH [#5174] were from Cell Signaling Technology. MST2 [#ab52641] was from Abcam, and p110 (pT1061) polyclonal antibody was developed and purified by Cell Signaling Technology. HRP-conjugated secondary antibody was used at 1:5000 in 5% milk and membrane were developed using ECL solution and exposed to film. 7.5% phos-tag gel was used to resolve phosphor-p110a (T1061) (Wako Diagnostics/Chemicals # 192–18001).
Lin T.Y., Ramsamooj S., Perrier T., Liberatore K., Lantier L., Vasan N., Karukurichi K., Hwang S.K., Kesicki E.A., Kastenhuber E.R., Wiederhold T., Yaron T.M., Huntsman E.M., Zhu M., Ma Y., Paddock M.N., Zhang G., Hopkins B.D., McGuinness O., Schwartz R.E., Ersoy B.A., Cantley L.C., Johnson J.L, & Goncalves M.D. (2023). Epinephrine inhibits PI3Kα via the Hippo kinases. Cell reports, 42(12), 113535.
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