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Cell lysis for peptide binding assay


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#1 jackysuen

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Posted 19 October 2004 - 03:52 PM

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

#2 Nuchy

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Posted 20 October 2004 - 06:24 AM

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

#3 sunmore

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Posted 22 October 2004 - 12:40 AM

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

#4 orexin2003

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Posted 23 October 2004 - 01:25 AM

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.




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