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I'm trying to immobilize an antibody onto the glass surface by means of GPTOMS (glycidoxypropyl trimetoxy silane). Using the same protocol (silanization in toluene or acidified ethanol (see below)) I get different results any time, what are key points to check?
Looking through literature on immobilization, I see a lot of articles where authors use APTES (aminopropyl triethoxy silane) and only a few using GPTOMS, why? Epoxy group seems so atractive…
For toluene:
- Rince slides with water
- Ultrasonicate them in isopropyl alcohol for 3 min, dry them
- Put for 3 minutes into piranha solution
- Rinse with distilled water
- Treat with oxygen plasma for 3 min
- Slowly put slides into 2 or 4 % silane solution in toluene, incubate for 3 min
- Rince with toluene and isopropyl alcohol.
- Put them on dry heater (max temp. of ours is 97 C, most protocols say 110 C), incubate an hour
For ethanol:
- Rince slides with water
- Ultrasonicate them in isopropyl alcohol for 3 min, dry them
- Put for 3 minutes into piranha solution
- Rinse with distilled water
- Treat with oxygen plasma for 3 min
- Put slides slowly into 2% silane in ethanol solution (silane + alcohol + acetic acid (pH 4,5-5,5)), incubate 3 minutes
- Rince slides with ethanol
- Put them on dry heater (max temp. of ours is 97 C, most protocols say 110 C), incubate an hour
