I am trying to adopt a step by step protein carbonylation lab protocol by Colombo et al., 2016, for my biological samples, but unfortunately pellets aren't compleletly dissolving after adding 1ml of 6 M guanidine hydrochloride, despite 30 minutes incubation at 37 degree.
Hello Kamati ! Dry the pellet of solvent completely before dissolving and incubate, with shaking, in higher temeprature ( ex. 60*C overnight) with the tubes covered, spin the tubes before reading. The signal is stable. I measured the samples on Nanodrop with good precision. Article Promising effects of xanthine oxidase inhibition by allopuri...
Adam Figarski , Thankx for advice, but how did you dry your samples, vaccum dry or air dry? and incubate with shaking might be a little challange with my incubator.
After decanting last, clear wash with solvent I just put it into shaking waterbath I regularly use for incubations, set to 60*C. Not all samples like to dissolve fully
Hi Adam Figarski , when you run your samples for protein carbonyl, what did you use as a positive control or standard for the DNPH assay?
I am currently using a molar coefficient of 22 000 M/cm, but I feel it would be much better if I get a positive control or a standard...
Thankx....
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