I've been infecting T24 cell lines by bacterial samples. I know that to do Cytokine ELISA I have to centrifuge it.
I am asking what condition would be appropriate for Centrifugation?
Some are saying 1500 rpm, some recommends 1000g , and some 5000 rpm.
You may spin down at 10,000 g which will pellet almost all the bacteria. If some is still left you may do a quick antibiotic treatment appropriate for the bacteria or do a filtration with 0.22um filter tubes. This will remove all the bacteria from your samples and leave your cytokines in the supernatant/filtrate.
Sudeep Kumar sir, I did a centrifuge at 10000 rpm before for 10 minutes at room temperature, but couldn't able trace out IL-6 concentration after doing ELISA.
There is no way 10,000g would affect the IL6. Something else is wrong.
It's plausible that many cells would make IL-6 in response to bacterial infection. This should work. Although there are probably also cell types that don't. It's hard to tell from here where your problem might be. Feel free to post details.
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