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Western Blotting Sample Preparation Optimization
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 kits
Western Blotting Protein Quantification Optimization
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 silver
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 size
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 method
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 buffer
Western Blotting Antibody Concentration Optimization
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 concentration