Cell lysates were prepared in Laemlli Buffer. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transfer to PVDF membranes were performed for antibody incubations. Enhanced chemiluminescence (ECL) was used to detect proteins with West-Pico or West-Dura reagents (Pierce-Thermo). ECL imaging was performed on a Syngene G-Box XT4 system with the GeneSys imaging software. The following antibodies were used for Western blotting: PARP, Axin2, p-AMPK, t-AMPK, Caspase 8, pS6, S6, pERK, ERK, pS6K, S6K, pAKT, AKT, V5, pTAK1, TAK1, pSAPK/JNK (Cell Signaling), panRAS (BD Biosciences); ERK, GAPDH (Santa Cruz). All antibodies were diluted in 2% BSA/TBS-T solution. Secondary anti-mouse or rabbit HRP-conjugated antibodies were purchased from Cell Signaling.
Panras
Manufactured by BD
PanRAS is a laboratory equipment product developed by BD. It is designed to detect and identify RAS gene mutations. The core function of PanRAS is to analyze genetic samples and provide information about the presence and type of RAS gene mutations.
Automatically generated - may contain errors
Lab products found in correlation
P sapk jnk, by Cell Signaling Technology (1 mentions)
P tak1, by Cell Signaling Technology (1 mentions)
G box xt4 system, by Syngene (1 mentions)
Gapdh, by Abcam (1 mentions)
Protease and phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Rabbit anti phospho yap ser127, by Cell Signaling Technology (1 mentions)
P16 ink4a, by Proteintech (1 mentions)
Mouse anti yap taz, by Santa Cruz Biotechnology (1 mentions)
2 protocols using panras
1
Western Blot Analysis of Signaling Proteins
2
Immunoblotting of Cell Signaling Proteins
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Cells were lysed in modified RIPA buffer supplemented with protease and phosphatase cocktail inhibitors (Sigma). Whole cell lysate were separated via SDS-PAGE electrophoresis and transferred to 0.2 μM PVDF Western blotting membrane. Membranes were then probed with mouse anti-Yap/Taz (Santa Cruz, 101199, 1:100), rabbit anti-phospho Yap Ser127 (Cell Signaling, #9411, 1:1000) (this antibody can detect murine pYap S112 and pTaz S8925 (link)), pan-Ras (BD Bioscience, #610001, 1:1000), p53 (Cell Signaling, #2524, 1:1000), p16-INK4A (Proteintech, #10883-1-AP, 1:500), small-T-antigen (Millipore, #D01, 1:1000), tubulin (Sigma, #B512, 1:2000), actin (Sigma, #A5060, 1:2000) and Gapdh (Abcam, #Ab8245, 1:10000). Primary antibody binding was then visualised using species-specific conjugated secondary antibodies (Invitrogen) and digital imaging.
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