Thiol has the tendency to oxidized and form dimer in the solution. Thiol and chemicals are incubated at room temperature followed by LC-MS analysis, i found that the dimerization signals are too high. how can i reduce the dimerization effect? or is there any alternative reagent to reduce the dimer effects? Any suggestion or advice would be helpful.
Keep your solution under Ar or nitrogen. Make you solution more acidic.
Dear Rahul Upadhyay Nepal
you can use 2 strategies:
1) Remove oxigen from buffers by degassing with nitrogen flow and afterwards working under an anaerobic atmosphere (eg in a glove box)
2)If for free thiol do you mean free cisteines in a protein you can block it with a reagent as iodoacetamide, AMS or others.
3) Theorerically you can try also to add low amount of week reducing agents (eg GSH/GSSH) but the formation of sone S-S is even possible
good luck
Manuele
Rahul Upadhyay Nepal
Are you measuring thiols?
If yes, then
Manuele Martinelli Your point 1, see my response.
Your point 2 might be useful. The idea is to convert thiol to a stable product. You can oxidize your thiol to RSSR and then measure RSSR. However, your sample may contain the dimer and thiol=RSSR-RSSRo.Thus, you need to know RSSRo
Your point 3. You must add an excess of GSH
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