RIPA lysis buffer (Beyotime, China) was used for the extraction of total proteins, and the determination of protein content was carried out with a BCA protein assay kit (Pierce, USA). The protein separation was performed on an 8-10% polyacrylamide gel by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes. Chemiluminescence was used to visualize the proteins after they had been incubated with secondary and primary antibodies. The bands were quantified using the Imaging J analysis software. The detailed protocol was described previously 25 (link), 26 . Following antibodies used in this study: anti-LATS1 (dilution 1:1000; #17049-1-AP, Proteintech), anti-Gli1 (dilution 1:1000; #ab217326, Abcam), anti-YAP1 (dilution 1:1000; #14074, Cell Signaling Technology (CST)), anti-p-mTOR (dilution 1:1000; #9205, CST), anti-mTOR (dilution 1:1000; #2983, CST), anti-GAPDH (dilution 1:5000; #AG019, Beyotime).
Anti lats1
Manufactured by Proteintech
Anti-LATS1 is a laboratory reagent used for the detection and quantification of LATS1 (large tumor suppressor 1) protein in various samples. LATS1 is a serine/threonine-protein kinase that plays a crucial role in the Hippo signaling pathway, which is involved in the regulation of cell proliferation, apoptosis, and organ size.
Automatically generated - may contain errors
Lab products found in correlation
Anti yap1, by Cell Signaling Technology (1 mentions)
Anti gli1, by Abcam (1 mentions)
Anti gapdh, by Beyotime (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Anti p p130cas, by Cell Signaling Technology (1 mentions)
Anti rabbit igg, by Cell Signaling Technology (1 mentions)
Anti n cadherin, by Cell Signaling Technology (1 mentions)
Anti β catenin, by Cell Signaling Technology (1 mentions)
Anti e cadherin, by Cell Signaling Technology (1 mentions)
Anti limk, by Cell Signaling Technology (1 mentions)
Anti pak, by Cell Signaling Technology (1 mentions)
Anti yap, by Proteintech (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Anti cortactin, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Pvdf membrane, by Pall Corporation (1 mentions)
Enhanced chemiluminescence detection kit, by Merck Group (1 mentions)
Anti α tubulin antibody, by Proteintech (1 mentions)
Anti lats2, by Proteintech (1 mentions)
Fluorchem fc2 multi imager 2, by Bio-Techne (1 mentions)
3 protocols using anti lats1
1
Western Blot Analysis of Protein Expression
2
Western Blot of Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein from cell or tissue lysates was separated by SDS-polyacrylamide gel electrophoresis (PAGE) and electro-transferred onto a polyvinylidene difluoride (PVDF) membrane (Pall Corp, Port Washington, NY). Then the membranes were blocked with 5% skimmed milk and incubated using primary antibodies anti-CSRP2 (1:800 dilution, Sigma), anti-GAPDH, anti-p130Cas, anti-p-p130Cas (Tyr410), anti-PAK, anti-p-PAK, anti-LIMK, anti-p-LIMK, anti-cortactin, anti-p-cortactin, anti-E-cadherin, anti-N-cadherin, anti-vimentin, anti-β-catenin (Cell Signaling Technology, Beverly, MA), anti-LATS1, anti-Yap (Proteintech), anti-ERK1/2, anti-p-ERK1/2 (Thr202/Tyr204), anti-p-Yap (Ser127) (Absci), anti-p-LATS1 (Thr1079/1041) (Bioss) at 4 °C. Then the membranes were incubated with appropriate secondary antibodies (anti-rabbit IgG, 1:3000 dilution, #7074; cell signaling). Enhanced chemiluminescence (Pierce, Rockford, 1L, USA) was used to detect the signal.
3
Western Blot Analysis of Signaling Proteins
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Cells were lysed in ice-cold RIPA buffer containing a protease-inhibitor cocktail (Roche). Protein content was quanti ed with a BCA protein assay kit (Thermo Fisher Scienti c). About 30 μg of protein was subjected to electrophoresis, transferred onto PVDF membranes (Millipore), and blocked with 5% nonfat dry milk in Tris-buffered saline containing 0.1% Tween 20. Membranes were incubated overnight with the following primary antibodies: anti-AKT (dilution 1:1000; CST), anti-AKT1 (1:1000; CST), anti-p-AKT(Ser473) (1:1000; CST), anti-p-AKT (Thr308) (1:1000; CST), anti-β-catenin (1:2000; Proteintech), anti-ALDH1A1 (1:1000; Abcam), anti-Vimentin (1:500; CST), anti-MST1 (1:1000; Proteintech), anti-MST2 (1:1000; Proteintech), anti-LATS1 (1:1000; Proteintech), anti-LATS2 (1:1000; Proteintech), anti-p-LATS1/2
(1:1000; Absin), anti-YAP (1:1000; Ptoteintech), anti-p-YAP (1:1000; Absin). Anti-GAPDH antibody (1:3000; Proteintech) and anti-α-tubulin antibody (1:3000; Proteintech) served as endogenous controls. The speci c bands were visualized using secondary anti-rabbit or anti-mouse antibody (1:3000; Proteintech), enhanced chemiluminescence detection kit (Millipore), and FluorChem FC2 Multi-Imager II (Alpha Innotech).
(1:1000; Absin), anti-YAP (1:1000; Ptoteintech), anti-p-YAP (1:1000; Absin). Anti-GAPDH antibody (1:3000; Proteintech) and anti-α-tubulin antibody (1:3000; Proteintech) served as endogenous controls. The speci c bands were visualized using secondary anti-rabbit or anti-mouse antibody (1:3000; Proteintech), enhanced chemiluminescence detection kit (Millipore), and FluorChem FC2 Multi-Imager II (Alpha Innotech).
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