I find some cell lysis buffer (like 1%NP-40 lysis buffer) has buffer agent in it,the commonly used buffer agent is HEPES or TEOA.
Nowdays,I need to get the cell lysis to do the CUAAC reaction,Because I Want to study aspirin, I get the aspirin-alkyne probe as the papers do(Sci Rep.2015 Jan 20;5:7896,J. Am. Chem. Soc., 2013, 135 (39), pp 14568–14573.), I am worring some buffer agent in the lysis buffer will disturb my CUAAC reaction. whether they will react with proteins or compound,even if my compound has form a covalent bond with proteins?
So HEPES and TEOA,which one is more suitable ? Whether these two buffer agent will react with aspirin-alkyne or proteins?
From your diagram, it appears that your chemical probe reacts with the epsilon amino group of lysine. If so, then you will want to avoid using a buffer that contains a primary amine because the compound will react with it. Both triethanolamine and HEPES have tertiary amines, not primary amines. If the chemical reaction favors primary amines over tertiary amines, either buffer should be OK. To be absolutey sure that there will be no cross-reaction, you could use a non-amine buffer instead: sodium or potassium phosphate or sodium bicarbonate.
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