Good day
Kindly guide and advise me on whether I can use ELISA assay to quantitate Antifreeze protein( AFP) instead of western blot? Is it acceptable to use ELISA and is it considered more effective and reliable.
ELISA is easier, more sensitive and more accurate than a western blot. Do duplicates or triplicates for your samples so your results are more reliable.
Shilan s Saleem , ELISA and Western blotting are too of the most commonly used method in molecular biology for the detection and quantification of proteins. Both the methods are based on the principle of binding specific antibodies to target proteins.
You go with ELISA, because it is immunoassays carried out in 96- well plates as a single,rapid and highly sensitive method for measuring the specific protein in sample.
Their are some regions to use ELISA over western blot for quantification of antifreeze proteins.
1. For detection of very small amounts of target proteins/rare proteins/low abundance proteins...so it is mostly suitable for antifreeze proteins as compare to western blot...because western blot is used to quantitative /detect wide range of proteins.
2. Unlike ELISA , Western blot is a multi step process, that can take from several hours to overnight to perform. Western blotting is also less sensitive than ELISA.
If you are working with antifreeze proteins then quantifying it with ELISA.
Shilan s Saleem ELISA and WB are common techniques for proteins detection and quantification, ELISA is more rapid and more accurate, but you need to buy the kits form a reliable company.
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