The expression of TGFβR2 was assessed by Western blot analysis and GAPDH was used as an internal control. Total proteins were extracted from tissues and cultured cells that were solubilized in lysis buffer supplemented with a protease and phosphatase inhibitor cocktail on ice. Proteins were separated by 10% SDS‐PAGE and then transferred to PVDF membranes (Roche, Mannheim, Germany). The membranes were incubated for 2 hour with 2% BSA at room temperature for 1 hour and incubated overnight at 4°C with primary anti‐TGFβR2 (1:200; Santa Cruz Biotechnology) and anti‐GAPDH (1:5000; Santa Cruz Biotechnology). Subsequently, the membranes were washed and incubated with appropriate secondary antibodies. The immune complexes were visualized with an ECL Prime Western Blot Detection System (Thermo, EHVEGFACL, Carlsbad, CA, USA).
Ecl prime western blot detection system
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The ECL Prime Western Blot Detection System is a chemiluminescent detection reagent used for Western blot analysis. It is designed to detect and quantify proteins on membranes.
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Lab products found in correlation
2 protocols using ecl prime western blot detection system
1
Quantification of TGFβR2 Expression
2
Protein Expression Analysis by Western Blot
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For western blot analysis, cell lysates were prepared in RIPA lysis buffer containing protease inhibitors and phosphatase inhibitors. Proteins of different molecular weights were separated by SDS/PAGE and then transferred onto a polyvinylidene difluoride membrane, which was blocked with BSA or nonfat milk for 2 h. The membrane was then sequentially incubated with specific primary antibodies and secondary antibodies. We finally visualized it using the ECL Prime western blot detection system (Thermo Fisher, Shanghai, China).
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