Hi. I have a commercially available, widely used, ELISA kit for testing protein carbonyl content. This is from CellBioLabs. I'm wondering what lysis buffers others have used when doing homogenates from tissue samples? In general, what are good buffers for ELISA assays, and particularly for protein carbonyl content measurement. I have mouse retinas, relatively easy to homogenize.

I ran a couple of tests already, but I am not super convinced about the results. I got 2-3 fold higher protein carbonyl contents from WT samples that were lysed in PBS only (no any detergents, nothing) than with commercially available RIPA buffer.

I treat homogenates with Streptomycin sulphate, so there should not be nucleic acids.

Also, it seems that otherwise the method is working, calibration curve is okay etc, I am just buzzled why I get so much higher apparent protein carbonyl contents in samples lysed in pure PBS, than RIPA buffer.

I looked at like 20-30 referenced articles and no one seem to discretely enclose how they prepared samples for this analysis!!