I want to perform a whole cell colorimetric assay with hydrogen peroxide. For example DMP , RB 18 or ABTS assay. And I'm running into problems because I wanted to perform this in a whole cell system or in partially lyzed cells (by freezing cycles). But I cannot detect any colorimetric change. My protein should be in the periplasm. So the question is: is hydrogen peroxide able to pass through the outer membrane? Or does anyone has experience with colorimetric assays with hydrogen peroxide in whole cell systems?
Anja, you can make it work. Please see my previous paper re. fixing the cells, permeablize with saponin and do immunocytochemistry with HRP, H2O2 and DAB staining.
Bernard MA et al. 2001 Diminished levels of the putative tumor suppressor proteins EXT1 and EXT2 in exostosis chondrocytes. Cell Motil Cytoskeleton. 48(2):149-62.
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