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- Following SDS-PAGE, transfer proteins onto blotting membrane according to the manufacturer’s instructions. Check protein transfer by staining the blot with Ponceau S for 1 min, then completely destain the blot by washing with distilled water. Place blot into blocking solution for 2 hr at RT, or overnight at 4°C.
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We know that western blotting protocols vary depending on your particular laboratory set-up and reagents of choice. Here are a series of protocols, each explicit to a specific combination of reagents, detection modes, and laboratory equipment. Please use the filters below to select the specific protocol that best fits your needs.
- Western Blotting Sample Preparation Techniques | Bio-Rad
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- Western Blotting Transfer Techniques | Bio-Rad
Western Blotting Protocol Library Filter by your laboratory...
- Western Blotting Sample Preparation Techniques | Bio-Rad
View Our Detection of Phosphorylated Proteins by Western Blotting Protocol. Explore Our Full List of Recommended Antibody Protocols. View TidyBlot Reagent Protocol. This western blot protocol provides a general procedure for use with the majority of Bio-Rad reagents.
On This Page. Gel electrophoresis and protein transfer by electroblotting. Follow us on: * Detailed instructions for making gels in-house can be found in Sambrook et al . For standard denaturing and reducing conditions, mix sample 1:1 by volume with Laemmli 2x sample buffer. Heat to 100 o C for 5 minutes or 70oC for 10 minutes.
Learn more. Western Blotting Technique. The western blotting workflow involves the selection of appropriate methods, apparatus, membrane, buffer, and transfer conditions. This involves two phases: protein transfer to a membrane and detection of the membrane-immobilized protein.
Western blotting, also called immunoblotting, is a widely used and accepted technique to detect levels of protein expression in a cell or tissue extract. This technique measures protein levels in a biological sample through antibody binding to a specific protein of interest.
