We are trying to functionalize nanooxides with streptavidin to use them for detection.
You can coat the oxide particles with mercaptosilane and then couple it with maleimide activated streptavidin (from Pierce).
Dear Stefansson ,
Can u give me some more information or references in this aspect
There are lots of references on coating mercaptosilane with glass, but here is a procedure that I have used:
You can clean the glass in conc. H2SO4 for 6hr (or ON). Rinse the glass with copious quantity of diH2O and dry the glass. Treat the glass with 3% (3-mercaptopropyl)trimethoxysilane (Sigma) in acetone (or 100% methanol) for 2 hr and then rinse the glass with acetone and dry with N2. Don't keep the -SH coated glass exposed to air for an extended time.
Make a 10 microg/ml solution of maleimide activated streptavidin (from Pierce) in PBS and add the glass into that mixture. Let it react with stirring for 2 hr at RT and then separate the glass from that solution and rinse with diH2O.
Optional: Block all non-reacted SH goups with 1 mM iodoacetate (or some other small -SH reactive compound).
Notes: if there is any H2O in the acetone (or methanol) or if the glass is not fully dried you will see some silane react with the water and aggregate. The mercaptosilane stinks, so keep your labmates happy and do the reaction in a fumehood.
Good luck.
Thank you for your suggestion sir. But my substrate is not glass. The starting material is sio2 nanopartticle. i would like to know still the same procedure holds good even except the H2SO4 cleaning step.
It's just a cleaning step. You can use NaOH or H2O2 instead.
For such purpose we performed silanization, read this publication:
Article AFM study of complement system assembly initiated by antigen...
Chemical polymer will be also suitable:
Article Polymerization Model for Hydrogen Peroxide Initiated Synthes...
@ Steingrimur Stefansson : Thank you so much sir. It is very helpful.
@Almira Ramanaviciene thank you for giving me a differnt of the process.
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