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Our western blot protocol includes solutions and reagents, procedures, and useful links to guide you through your experiment. Reviewed December 14 2020. Western blotting is a technique that uses specific antibodies to identify proteins that have been separated based on size by gel electrophoresis.
General Protocol for Western Blotting Protein separation by gel electrophoresis 1. Load equal amounts of protein (20 μg) into the wells of a mini (8.6 x 6.7 cm) or midi (13.3 x 8.7 cm) format SDS-PAGE gel, along with molecular weight markers. 2. Run the gel for 5 min at 50 V. 3. Increase the voltage to 100–150 V to finish the run in about 1 hr.
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Here, you can find a collection of western blot recipes for commonly used protein electrophoresis and western blot buffers and stock solutions, and general western blotting protocols for chemiluminescent and fluorescent detection to guide you through your experiment.
Western blot is a technique that is very useful for protein detection as it allows the user to quantify the protein expression as well. This paper covered the protocol, the theory behind that protocol, and some troubleshooting techniques.
- Tahrin Mahmood, Ping-Chang Yang
- 10.4103/1947-2714.100998
- 2012
- N Am J Med Sci. 2012 Sep; 4(9): 429-434.
Western Blotting Protocol. Western blotting (WB) is widely used to analyze specific protein expression in cell or tissue extracts. At Cell Signaling Technology (CST) we understand that western blotting experiments are time consuming and that their success has a critical impact on your research progress.
Protein transfer and visualization in western blot. Detailed protocol for the transfer and staining of proteins in western blot. Includes visualization of proteins in gels, transfer, and development methods. Fluorescent western blot. Get started with fluorescent western blot with these quick tips.
