Hello, I am practicing GSH assay protocol (Rahman, 2006). I got the OD from the measurements, but I don't know how to express the results /mg of protein as the protocol mentions On the last table. Should I use Bradford assay?
Thank you
You can express as microgram GSH/mg of protein or micromoles of GSH/mg of protein. You can calculate the concentration of GSH in tissue by determine standard in sequence.
Expressing results as a GSH/mg protein is a normalization way in order to avoid differences caused by not using the same sample amount.
To express your results in µM/mg protein or nM/mg protein you should:
1. Perform a linear regression with standards to obtain the standard curve.
2. Interpolate your sample results in the standard curve to obtain sample concentration (in the same units of the standard curve. i. e., if the standard curve is in µM, your sample results will be in µM too).
3. Use a protein determination method or kit to obtain mg protein in each sample.
4. Divide the GSH concentration of each sample by mg of proteins of the same sample. In the last example, results will be in µM/mg protein).
I hope you'll find this information useful.
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