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Inquiry: Dear Sir/Madam My question is about cAMP Direct immunoassay kit (ab65355) and cGMP Direct immunoassay kit (ab65356) I want to know before adding 0.1 M HCl to cell culture (to break cells), I must pour cell culture media out and then add HCl or not. I lookforward to your reply. Best Regards
Asked on Jan 21 2013
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On page 7 the section b of assay protocol states "for suspension cells collect by centrifugation. Add 1 ml of 0.1M HCl for every 35 cm2 of surface area (e.g., 10 cm plate at 70% confluency is ˜110 cm2, so use ˜3.1 ml). Incubate at room temperature for 20 minutes on ice."
This means you need to centrifuge out the media. The volume to HCL can be calculated as per the instructions given.
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Answered on Jan 21 2013