N-hydroxysuccinimide(NHS) and EDC(1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide) are expected to be water soluble.But my solution gave semi-ester form with two immiscible portion of solid and water.If anyone is working with this system, please describe me the final form solution.
You should try EDC-NHS as 60mM and 40 mM respectively. The esters are stable only for 30-45 minutes so make sure you donot leave it to sit for longer than 30 minutes. I have tried it for immobilisation of Glucose oxidase on thiomalic acid and it works fine.
Amruta:
The final form of solution must be colourless. If you want to activate the surface to attach biomolecules then activate your surface with 400 mM EDC + 100mM NHSS. Normally in such reactions EDC activates the surface where as NHSS acts as a stabilizing agent preventing detoriation of EDC.
Please see SI file attached for the procedure. You will find the EDC-NHSS procedure under "Fabrication of SWCNT Immunosensors."
Thanks for the reply.I would like to know whether the reaction is temperature dependent?I am trying to activate the Au surface coated with MUA and MH in 1:10 (mM)ratio.
To the best of my knowledge this reaction is not temperature dependent. If you are using the Gold surface, please see this paper attached below from our group.
There is no role of temperature. You can thiolate your metal surface and afterward use the EDC:NHS. But make sure do not keep it for longer time. One precaution you can use, keep it in desiccator after adding EDC: NHS mixture.
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