Proteins were obtained by radioimmunoprecipitation assay lysis buffer (Abcam Corp, USA) and quantified by bicinchoninic acid. Protein, 50 μg per sample, was separated onto 10% SDS–PAGE gels and transferred onto a polyvinyl difluoride (PVDF) filter membrane. Then, PVDF membranes were incubated with 5% non-fat dried milk, anti-RRBP1 (Abcam Corp., USA), Smad-1 (Abcam Corp., USA), p-Smad-1 (Abcam Corp., USA), Smad-3 (Abcam Corp., USA), p-Smad-3 (Abcam Corp., USA), TGF-β1 (Abcam Corp., USA) and anti-GAPDH (Abcam Corp., USA), respectively. Finally, membranes were washed with phosphate-buffered saline (PBS) and incubated with secondary antibodies. Signals were developed using an enhanced chemiluminescence reaction kit (Applygen Technologies, Beijing, China).
Enhanced chemiluminescence reaction kit
Manufactured by Applygen
Sourced in China
The Enhanced chemiluminescence reaction kit is a laboratory product designed to facilitate the detection and analysis of proteins through chemiluminescence. The kit provides the necessary reagents and components to perform enhanced chemiluminescence reactions, which are commonly used in Western blotting and other protein-based assays.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Abcam (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Radioimmunoprecipitation assay lysis buffer, by Abcam (1 mentions)
P smad3, by Abcam (1 mentions)
Anti rrbp1, by Abcam (1 mentions)
Smad1, by Abcam (1 mentions)
P smad1, by Abcam (1 mentions)
Tgf β1, by Abcam (1 mentions)
Smad3, by Abcam (1 mentions)
Pvdf filter membrane, by Bio-Rad (1 mentions)
Ripa lysis buffer, by Abcam (1 mentions)
Anti smad1, by Abcam (1 mentions)
Anti p smad1, by Abcam (1 mentions)
Anti smad3, by Abcam (1 mentions)
Anti p smad3, by Abcam (1 mentions)
Anti tgf β1, by Abcam (1 mentions)
2 protocols using enhanced chemiluminescence reaction kit
1
Protein Expression Quantification Protocol
2
Western Blot Analysis of BZW1, Smad, and TGF-β1
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Proteins were extracted by radioimmunoprecipitation assay (RIPA) Lysis buffer (Abcam, Cambridge, MA, USA) and quantified by bicinchoninic acid (BCA) Protein Assay Kit (Thermo Scientific, Rockford, IL, USA). 40 µg of protein per sample was separated onto 10% SDS-PAGE gels and transferred onto polyvinyl difluoride (PVDF) filter membrane (Bio-Rad, Hercules, CA, USA). Then, PVDF membranes were blocked with 5% non-fat dried milk for 2 h, and incubated with anti-BZW1 (1: 1000, ABcam Corp, USA), anti-Smad1 (1: 1000, ABcam Corp, USA), anti-p-Smad1 (1: 1000, ABcam Corp, USA), anti-Smad3 (1: 1000, ABcam Corp, USA), anti-p-Smad3 (1: 1000, ABcam Corp, USA), anti-TGF-β1 (1: 2000, ABcam Corp, USA) and anti-GAPDH (1: 2000, ABcam Corp, USA), respectively at 4 overnight. Finally, membranes were incubated with secondary antibodies (horseradish peroxidase-conjugated anti-rabbit). Signals were developed using an enhanced chemiluminescence reaction kit (Applygen Technologies, Beijing, China).
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