Wanted - buffer systems for pH range 4 to 9 in cell culture expt.

Mr. P.A. Sansom psansom at hgmp.mrc.ac.uk
Thu Oct 6 05:47:24 EST 1994


Aim - test effect of pH on proteolysis of peptide by cell surface proteases

Method - 10^6 cells + peptide in 100 microlitre buffer at pH ranging from
4 to 9, incubated for 1 hour at 37C. Degradation assessed by RP-HPLC of
supernatant.

Requirements - overlapping buffer systems spanning pH range 4 to 9 that will
maintain cell integrity for duration of experiment. Some tests will include
inhibitors that can alter pH, thus buffering should be moderately strong.

Notes - I believe that low pH buffers can strip some proteins from the cell
surface - any information on this would also be appreciated.

Thanks in advance for any help that is offered.

Paul
psansom at hgmp.mrc.ac.uk

Paul A Sansom                                  tel: 081-748-9966 x4208
Department of Biochemistry                     fax: 081-748-5090
The Kennedy Institute of Rheumatology
6 Bute Gardens
Hammersmith
London W6 7DW



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