Can anyone provide the recipe in dissolving Bovine Serum Albumin in a buffered solution which can be used for cell sorting? I came across the solution: HBSS-BSA-EDTA medium but i'm a little confused as to the volume of HBSS that will be used in preparing the solution . I came across: HBSS (Ca2+/Mg2+ free), 1.0% BSA and 0.15% EDTA, pH 7.4. My questions are 1. At what volume will I prepare HBSS? 2. Will 1g of BSA and 0.15g EDTA be poured directly to the HBSS or will 1.0% BSA and 0.15% EDTA be prepared separately and then poured into HBSS?
The first question depends on the answer to the second. We make 20% BSA by dissolving 500g BSA in 500ml 1X PBS or in your case 1X HBSS followed by filtering through 0.22 or 0.45um filter top by vacuum, and make aliquots of 12.5ml each (and freeze the rest in -20C), so that when adding to 500ml bottle 1X PBS it will become 0.5% BSA in PBS, in your case, you can double that volume. For EDTA we buy 1M EDTA stock and you have to determine what the molar is for your 0.15%, then calculate based on 500ml total volume of HBSS, for example, we add 1ml of 1M EDTA (1mmole) to 500ml PBS to make 2mM EDTA. So the point is we add concentrated BSA and EDTA to a bigger volume of buffer to make the final concentration you want. Hope this is helpful.
Thank you for your answer Zhixin Jingq. I have a better understanding now.
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