- Table of Content
The first step to obtaining a clear, accurate Western blot is sample preparation. The critical step of sample preparation, the choice of protein extraction method, is a crucial one which ultimately makes the difference between a blank blot and a beautiful one. Boster supplies a range of lysis buffers and specialized extraction kits; use this guide to pick the right one for your task and start getting better results.
Keywords: Western blotting optimization, sample preparation, protein extraction kitsLearn More About Sample Preparation
One of the first issues to consider when conducting a total protein assay is the type of assay to use. A primary factor to consider in making your choice is the compatibility of the assay with the samples. The objective is to select an assay that requires the least manipulation or pre-treatment of the samples, and will give good results for your sample type. Use this comparison chart to help decide which protein assay is right for you
Keywords: Western blotting optimization, total protein analysis, protein quantification assay, BCA, commassie, protein silverLearn More About Protein Quantification
In order to obtain picture-perfect separation, the gel concentration and running conditions must be tuned to the protein being investigated. Optimize your gel concentration based on the size of the protein you are trying to resolve. Use this handy table to aid your estimates:
Keywords: Western blotting optimization, SDS-PAGE, gel percentage, gel concentration, pore sizeLearn More About SDS-PAGE
The biggest differences between transfer types arise in the type of equipment needed and the speed and efficiency of the transfer. Pick your transfer method carefully to get the best results in your own time and for the right price.
Keywords: Western blotting optimization, protein transfer, wet transfer, semi-dry transfer, dry transfer, best transfer methodLearn More About Protein Transfer
To obtain the best results from your western blot, you must decide which type of blocking buffer to use based on the detection system used and the antigen you’re trying to detect. Use this guide to decide which blocking buffer is best suited to your application:
Keywords: Western blotting optimization, blocking buffer, bsa, nonfat milk, pbs, tbs, best blocking bufferLearn More About Membrane Blocking
No single antibody or antigen concentration will work for every experiment. In order to obtain the most perfect Western blots, the antibody concentration needs to be optimized. The ideal antibody concentration is dependent on the concentration of antigen, the specificity and affinity of the antibody, and experimental conditions such as buffer composition. Use this guide to help optimize your antibody concentration and get better results:
Keywords: Western blotting optimization, antibody concentration, dot blot assay, best antibody concentrationLearn More About Antibody Concentration
Learn the concept behind Western blotting. It is a technique that is used to detect specific proteins in the given sample. It usually involves two major processes, namely, SDS-polyacrylamide gel electrophoresis and protein blotting and testing.
Learn ELISA Principle
Check out this Western Blot sample preparation guides to learn how to get the best results from your sample type. Learn more about sample preparation in this guide.
Learn Western Blot Sample Preparation
Get to learn Pro tips on resolving common Western Blot issues such as weak signal, wrong band size, smiley gel, and high background.
Browse Western Blot Troubleshooting Tips