PBS buffer for cell isolation sing dynabeads - (Apr/17/2008 )
I'm isolting CD133+ cells using dynabeads. According to the protocol, I need to prepare the folloing PBS buffers and would like help in understanding the requirements. firstly I have already prepared 1X PBS (pH 7.4), autoclaved and now I have to prepare teh following:
1) PBS with 0.1% BSA, pH 7.4
question: when I add 0.2g of BSA to 200ml prepared PBS, do I have to autoclave again?
2) PBS with 0.1% BSA and 0.6% Na-Citrate or 2mM EDTA.
question: I will just add the 0.6% Na-Citrate or 2mM EDTA to 100ml of BS prepared in step 1. Do I have t autoclave again?
do not autoclave with bsa. it will denature (and probably form clumps).
you can filter sterilize and should if the protocol requires you to maintain sterility.
Thank you very much