MDA-MB-231 and MCF-7 cells were harvested, washed, and lysed with RIPA buffer (Beyotime, China) containing protease inhibitor cocktail (Roche) for 30 min on ice. Total protein was extracted, and quantitated by a BCA kit (Beyotime, China) according to the manufacturer’s instruction. The protein samples were separated by electrophoresis in 10%~ 12% gel, and then transferred onto PVDF membranes (Millipore). Blotted membranes were blocked and incubation with primary antibodies overnight at 4 °C. The membranes were washed 5 min for 3 times with TBST, and subsequently incubated 1 h with HRP-linked secondary antibody (Cell Signaling Technology, USA) at room temperature. The band intensities were visualized and detected by an enhanced chemiluminescence detection system (Bio-Rad Laboratories). Primary antibodies against c-Myc, E-cadherin, N-cadherin, Vimentin, GLUT1, LDHA and β-actin were obtained from Cell Signaling Technology. Primary antibodies against PC4 were obtained from Sigma.
Primary antibodies against pc4
Manufactured by Merck Group
Primary antibodies against PC4 are laboratory reagents designed for use in various research applications. These antibodies specifically target and bind to the PC4 protein, which plays a role in transcriptional regulation. The core function of these primary antibodies is to facilitate the detection and analysis of PC4 in biological samples, supporting researchers in their investigations.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (4 mentions)
Protease inhibitor cocktail, by Roche (4 mentions)
Ripa buffer, by Beyotime (4 mentions)
Bca kit, by Beyotime (4 mentions)
Enhanced chemiluminescence detection system, by Bio-Rad (4 mentions)
Hrp linked secondary antibody, by Cell Signaling Technology (3 mentions)
Glut1, by Cell Signaling Technology (3 mentions)
β actin, by Cell Signaling Technology (3 mentions)
C myc, by Cell Signaling Technology (2 mentions)
4e bp1, by Cell Signaling Technology (2 mentions)
Hif 1α, by Cell Signaling Technology (2 mentions)
Bcl2 associated x (bax), by Cell Signaling Technology (2 mentions)
Bcl 2, by Cell Signaling Technology (2 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
P p70s6k, by Cell Signaling Technology (1 mentions)
Primary antibodies against β actin, by Cell Signaling Technology (1 mentions)
T mtor, by Cell Signaling Technology (1 mentions)
T 4e bp1, by Cell Signaling Technology (1 mentions)
4 protocols using primary antibodies against pc4
1
Protein Expression Analysis in Breast Cancer Cell Lines
2
Protein Expression Analysis in Pancreatic Cancer Cell Lines
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CFPAC-1 and AsPC-1 cells were harvested, washed, and lysed with RIPA buffer (Beyotime) containing protease inhibitor cocktail (Roche) for 30 min on ice. The total protein was extracted and quantified using a BCA kit (Beyotime) according to the manufacturer’s instructions. The protein samples were separated by electrophoresis using a 4%~20% gel and then transferred onto PVDF membranes (Millipore). The blotted membranes were blocked and incubated with primary antibodies overnight at 4°C. After 3 washes with TBST and a 1-h incubation with an HRP-conjugated secondary antibody (Proteintech) at room temperature, the membranes were developed using an enhanced chemiluminescence detection system (Bio-Rad Laboratories) to detect the protein. The primary antibodies against PC4 were obtained from Sigma, while the primary antibodies against β-actin, p-mTOR, t-mTOR, p-p70s6k, t-p70s6k, p-4EBP1, t-4EBP1, CDK4, CDK6, Cyclin D and Cyclin E were obtained from Cell Signaling Technology.
3
Western Blot Analysis of Apoptosis and Autophagy
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The cell lines were harvested, washed, and lysed with RIPA buffer (Beyotime, Nanjing, China) containing protease inhibitor cocktail (Roche, Basel, Switzerland) for 30 min on ice. Total protein was collected and quantitated by a BCA kit (Beyotime, Nanjing, China) according to the recommended instruction. The protein samples were separated by electrophoresis in gel and then transferred onto PVDF membranes (Millipore, Massachusetts, USA). Blotted membranes were incubated with primary antibodies overnight at 4°C. The membranes were washed three times with TBST for 5 min and then incubated with HRP-linked secondary antibody (Cell Signaling Technology, Houston, USA) for 1 hour at room temperature. Band intensities were detected and visualized by an enhanced chemiluminescence detection system (Bio-Rad Laboratories, Hercules, CA). Primary antibodies against c-Myc, LC3, SQSTM1, ATG7, poly ADP-ribose polymerase (PARP), CASPASE3, Bcl-2, BAX, PI3K, S6K1, AKT, mTOR, 4EBP1, AMPK, P38, P53, HIF-1α, GLUT1, PKM2, HK2, LDHA, and β-actin were obtained from Cell Signaling Technology. Primary antibodies against PC4 were obtained from Sigma.
4
Western Blot Analysis of Cellular Proteins
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The cell lines were harvested, washed, and lysed with RIPA buffer (Beyotime, China) which contain protease inhibitor cocktail (Roche) for 30min on ice. Total protein was collected and quantitated by a BCA kit (Beyotime, China) according to the recommended instruction. The protein samples were separated by electrophoresis in gel, and then transferred onto PVDF membranes (Millipore). Blotted membranes were incubated with primary antibodies overnight at 4°C. The membranes were washed 5min for 3 times with TBST, and then incubated with HRP-linked secondary antibody (Cell Signaling Technology, USA) 1h at room temperature. The band intensities were detected and visualized by an enhanced chemiluminescence detection system (Bio-Rad Laboratories). Primary antibodies against c-Myc, LC3, SQSTM1,ATG7,PARP,CASPASE3,Bcl-2,BAX,PI3K,S6K1,AKT,mTOR,4EBP1,AMPK,P38,P53,HIF-1α,GLUT1,PKM2,HK2,LDHA and β-actin were obtained from Cell Signaling Technology. Primary antibodies against PC4 were obtained from Sigma.
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