Tissues and cells were lysed in RIPA buffer (cat#V900854, Sigma, MO, USA) and protein concentration was measured with the BCA method (cat#P0012, Beyotime, Shanghai, China). Proteins were isolated by SDS-PAGE, transferred to PVDF membranes (Millipore, Billerica, USA), incubated with primary and secondary antibodies and imaged with the ECL detection reagent (cat#12630, CST, California, USA) using a ChemiDoc™ XRS+ System. The primary antibodies included anti-Mus81 (1:1,000; cat#ab14387, Abcam, Cambridge, UK), anti-BRD4 (1:1,000; cat#ab128874, Abcam), anti-ZEB1 (1:1,000; cat#ab180905, Abcam), anti-E-cadherin (1:1,000; cat#3195, CST, California, USA), anti-N-cadherin (1:1,000; cat#13116, CST), anti-Snail1 (1:1,000; cat#9782, CST), anti-ZEB2 (1:1,000; cat#14026-1-AP, Proteintech, Wuhan, China), anti-Sirt5 (1:500; cat#15122-1-AP, Proteintech), anti-GAPDH (1:5,000; cat#G9545, Sigma). The secondary antibodies included HRP conjugated goat anti-Rabbit (1:3,000; SA00001-15, Proteintech) and anti-Mouse (1:3000; cat# SA00001-1, Proteintech).
Anti zeb2
Manufactured by Proteintech
Sourced in China
Anti-ZEB2 is an antibody product used to detect the ZEB2 protein in various experimental applications. ZEB2 is a transcription factor involved in the regulation of gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Anti brd4, by Abcam (1 mentions)
Anti zeb1, by Abcam (1 mentions)
Anti mus81, by Abcam (1 mentions)
Anti sirt5, by Proteintech (1 mentions)
Sa00001 15, by Proteintech (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Anti gapdh, by Merck Group (1 mentions)
Bca method, by Beyotime (1 mentions)
Anti gadph, by Proteintech (1 mentions)
Anti e cadherin, by Proteintech (1 mentions)
Anti n cadherin, by Proteintech (1 mentions)
Ecl reagent, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
3 protocols using anti zeb2
1
Protein Expression Analysis by Western Blotting
2
Western Blot Protein Expression Analysis
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Cells were lysed with RIPA cell lysis buffer in the presence of protease inhibitor cocktail (Sigma, USA). The same amount of protein samples were loaded onto 10% SDS-PAGE and then transferred onto PVDF membranes. After blocked by skim milk, the membranes were incubated in the primary antibodies diluted by TBST buffer for overnight at 4°C and then in the HRP-conjugated secondary antibody for 2 h at room temperature. Finally the protein bands images were captured by a Tanon detection system with ECL reagent (Thermo). The primary antibodies used in the experiments were Anti-GADPH, anti-E-cadherin, anti-N-cadherin, and anti-ZEB2 antibodies were purchased from Proteintech.
3
Western Blot Analysis of Protein Targets
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Total proteins were extracted using RIPA lysis buffer supplemented with a protease inhibitor cocktail, as previously described [19 (link), 20 ]. Proteins were separated by SDS-PAGE and then blotted onto PVDF membranes. The membranes were incubated with primary antibodies overnight at 4 °C and then incubated with secondary antibodies for 1 h at room temperature. Finally, the proteins were detected by chemiluminescence according to the manufacturer’s recommendations (Advansta, CA, USA). The following primary antibodies were used: anti-RAB14 (1:1000, Proteintech, Wuhan, China), anti-ZEB2 (1:1000, Proteintech, Wuhan, China), anti-SOX4 (1:1000, Millipore, Billerica, MA, USA), and anti-GAPDH (1:2000, Sungene, Tianjin, China).
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