After color development reaction in ELISA we use acid to stop the reaction, but color must be read immediately, otherwise color intensity decreases proportionally, so it may destroy calibration curve structure. Is there any way to stabilize color for a longer period of time?
I mean can we use any additive to acid to do stabilize color?
For how long do you need to stabilize the color of your solutions? Normally there are STOP solutions that keep stable your reaction for an hour or so.
Using 2M H2SO4 as stop solution, I never had a problem to be able to walk from my bench to the spectrometer. To avoid influence from TMB vs light, you could wrap your (96-well) plate in aluminium foil. Unless we talk of course about letting it stand for several hours?
Use of heavy salt solutions such as nickel chloride, strontium chloride or manganese chloride to the substrate solution stabilizes the color. You may include these at about 1 mm concentration. Develop enzymatic reaction in a dark place! The stop solution [Acid solution] is inevitable and need to be used to terminate the HRP further reaction.
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