I want to quantify the Angiotensin-converting enzyme (ACE) in cell cultured samples. I prefer to do ELISA to quantify ACE.
Which lysis buffer should be used to lyse the cells? The supernatant collected post-lysis should not effect the ELISA experiment.
Hi Chandan,
Usually when I had to proceed to an ELISA starting from cells I used to lyse them by several freeze-thaw cycles. The best is to use liquid nitrogen and hot water bath to make fast cycles. At least 3 or 4 cycles and then you can start your ELISA, but you must be careful to wash very well at each step of your ELISA so that, as you said, what your recuperate don't interfere with the experiment.
Let me know how it's goin on.
Best of luck.
My question is what compartment you want to measure? If you want to mneasure ACE released into the medium then there is not question about lysis buffer. If you want to measure ACE inside of the cells then I see no problem using any kind of lysis buffer as long as the detergent will not intefere with your asay kit. Usually the commercial assay kit will instruct you what kind of lysis buffer to use.
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