After SDS-PAGE electrophoresis, the proteins were transferred to a nitrocellulose membrane. Then the membrane was blocked in 5% skimmed milk for 1.5 h at room temperature. After washing three times with TBST, the membranes were incubated with primary antibodies against E-cadherin (Santa Cruz, sc-8426, 1:1000), α-SMA (Santa Cruz, sc-53142, 1:1000), vimentin (Santa Cruz, sc-6260, 1:1000), Smad7 (Santa Cruz, sc-365846, 1:500), TGFβ1 (Santa Cruz, sc-130348, 1:500), TGFβRII (Santa Cruz, sc-17799, 1:500), p-Smad2 (Abcam, ab188334, 1:1000), p-Smad3 (Abcam, ab63403) and GAPDH (Santa Cruz, sc-365062, 1:2000) at 4°C overnight. The membrane was incubated with horseradish peroxidase (HRP)-bound secondary antibody (1:5000) for 1.5 h at room temperature, and visualized with BeyoECL Moon chemiluminescence kit (Beyotime, P0018FS).
Beyoecl moon chemiluminescence kit
Manufactured by Beyotime
Sourced in China
The BeyoECL Moon chemiluminescence kit is a laboratory equipment designed for the detection and quantification of proteins in Western blot analysis. The kit utilizes chemiluminescent substrates to generate a luminescent signal upon reaction with the target protein, allowing for sensitive and accurate protein visualization.
Automatically generated - may contain errors
Lab products found in correlation
α sma, by Santa Cruz Biotechnology (1 mentions)
Vimentin, by Santa Cruz Biotechnology (1 mentions)
Tgf βrii, by Santa Cruz Biotechnology (1 mentions)
Tgf β1, by Santa Cruz Biotechnology (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
Radioimmunoprecipitation assay lysis buffer, by Beyotime (1 mentions)
Bovine serum albumin (bsa), by Solarbio (1 mentions)
Phosphatase inhibitor cocktail, by MedChemExpress (1 mentions)
Pmsf reagent, by Beyotime (1 mentions)
Amersham imager, by Cytiva (1 mentions)
2 protocols using beyoecl moon chemiluminescence kit
1
Western Blot Analysis of EMT Markers
2
Western Blotting Protein Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was extracted using radio-immunoprecipitation assay lysis buffer (Beyotime, Shanghai, China) containing 1% PMSF reagent (Beyotime, Shanghai, China) and a 1% phosphatase inhibitor cocktail (MedChemExpress, Shanghai, China). Denatured total proteins were electrophoresed on a 12% polyacrylamide gel and transferred onto polyvinylidene fluoride membranes. Blocking was performed for 1 h in 5% bovine serum albumin (Solarbio, Beijing, China), and membranes were incubated overnight at 4 °C with the indicated rabbit anti-human antibodies (1:3000). Subsequently, the membranes were incubated with HRP-conjugated goat anti-rabbit secondary antibodies (1:5000) for 1 h. The blots were visualised using the BeyoECL Moon chemiluminescence kit (Beyotime, Shanghai, China) on the Amersham Imager 680 imager. Grey values were analysed using the ImageJ software. Band densities were normalised to that of β-actin or GAPDH.
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