Protocol Online logo
Top : Forum Archives: : Molecular Biology

PBS buffer for cell isolation sing dynabeads - (Apr/17/2008 )

Hi,

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?

Thanks
SF

-Sarwat-

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.

-mdfenko-

Thank you very much

-Sarwat-