I want to attach some chemical molecules to my substrate, and I need to use the EDC/NHSS coupling reaction between the amino group and the carboxyl group. However, my first experiment failed, so I would like to ask what pH and temperature conditions should I use for the EDC/NHSS coupling reaction between the amino group and the carboxyl group? And is there any other details I should pay attention to?
PH 5 in MES buffer is usually a good place to start, but the molar proportions of the carrier and the molecules is a key. I would recommend to try several conditions and see of you get anything from it
I normally use the 2-step EDC/NHS method. Assuming that your chemical has the carboxylate group and the substrate has the amine groups. Carry out the EDC/NHS activation step of your chemical in 50mM MES/NaOH buffer, pH 4.7 ( without sodium azide) then add your activated chemical to your substrate in 0.1M sodium bicarbonate, pH 8.1-8.3
My base is a porous film with carboxyl groups on the surface and I want to graft some functional groups with amino groups onto it. I activate the carboxyl groups in MES buffer at room temperature, pH=5.5, and then binding the product to the amino group in PBS buffer at pH=7.2.
The article I read has two statements, the EDC/NHS method and the EDC/NHSS method. Could there be any difference? Ahmed Jehanli
The difference between the two is that NHSS has a sulfo group making it more water soluble. Good to use if your compound is very sensitive to using dimethyl formamide or dimethylsulfoxide to dissolve NHS.
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