Hello,
I am trying to determine the cysteine and methionine content from soil extracted ACE-proteins using Ellman's reagent. I have gone over many protocols and all of them use the same sodium phosphate solution (0.1 M sodium phosphate, 8.0 pH). I don't know if it would be compatible to replace this reaction buffer with the same buffer for extracting the soil proteins (20 mM sodium citrate, 7.5 pH)? I am also developing the protocol for a 96 well plate and if anyone has guidance, I would really appreciate it.
I think it will be OK, but the absorbance signal may be somewhat less stable than with phosphate buffer. This is based on an experience I once had where the signal was less stable using MOPS buffer instead of phosphate.
I suggest you prepare the Ellman's reagent in phosphate buffer even if the samples are in citrate buffer.
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