Cell lysis for peptide binding assay - (Oct/19/2004 )

Hi everyone,

I'm doing a peptide binding assay where I need to lysis cells and expose a chemical in the cells after initial binding. I don't really need to reserve any proteins as I'm interested in the chemicals and it should be pretty tough (well, at least the chemist told me so). So does that mean I can just use simple detergent to do the job? Can anyone give me some suggestion please?

Millions thanks,
j..s

Let try to use sonicator lysate cell. This way is very easy to complete lysis cell.

lysing the cells with 0.1N HCl would be preferred for any other detergent even though it should n't be an issue with your assay.

sunil

Well i had worked in the past on tumor cell lines both adherent and suspension cell lines looking at best method for cell lysis to compare the release of one intracellular enzymes. I tried detergent lysis, heat treatment, sonification and alternate freeze thawing. I found that alternate freezing cells in liquid nitrogen followed by thawing on water bath at 56 Degree centigrade for 4-6 times results in 100% cell lysis. i would strongly recommend you to try this method it is very simple method, if cold shock is not an issue with your assay procedure.