I am looking for a way to measure GSH in hemolysed blood samples. I have found a protocol from Browne and Armstrong (Methods in Molecular Biology, vol 108), using o-phthalaldehyde and N-ethylmaleimide for fluorometric detection of GSH.
My problem is that this protocol is a test-tube protocol; it requires larger volumes and a spectrometer capable of reading quartz cuvettes. In our lab, we only have a microplate spectrometer.
I was inspired by the Pierce BCA assay from ThermoScientific; their assay can be used for both test-tube protocol and microplate. Is it possible to design a similar assay for my GSH assay?
You mentioned that the method was fluorometric. Does your plate reader have fluorescence capability? If it is only capable of absorbance measurements, then the method with Ellman's reagent is more suitable. Either one can be done in a microplate format.
Hello Ivana, if your microplate reader can measure fluorescence at the same conditions like the spectrophotometer, it should work.
I agree with Adam, assay based on Ellman´s reagent is quite easy and it works well in the both cuvettes and microplates. You can find protocol for the assay in e.g.: Drug and Chemical Toxicology 2011; 34(3): 255-260
Hi Ivana,
I see two problems you might run into:
If you don't run into problems with the hemolysis, and the limit of detection is no problem...this should be possible. Good luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biochemical Techniques