Dear Andreas,

you actually gave me a wonderful idea with the "in beads" digestion, I have already seen similar protocols, maybe this can be the chance to do something new. I have already used Laemmli sample buffer and heating, but the yield never really satisfied me. I also would like to try something like Holmberg (Electrophoresis 2005, 26, 501–510) made: non ionic aqueous solutions at 70°C ... and beads not destroyed. We''ll see which idea will turn out to be the best one.

Sowieso ... vielen Dank für deine sinnvolle Hilfe/Empfehlungen, und gerne wünsche ich dir ein schönes Wochenende! 

Best

Roberto

Hi Roberto, 

you can release proteins from beads using 70% formic acid. Then you can either exchange the buffer and perform in solution digestion as Andreas suggested or separate protein on SDS gel followed by in gel digestion. 

I will also try this approach,

Thanks for helping!

Hi Roberto,

Did you find the best way to elute proteins from dynabeads? I'm trying to biotinylate membrane proteins followed by isolation using strep-dynabeads. My yield is very low and I'm actually only getting about 10% membrane proteins and rest are cytoplasm, nuclear.

I have been using heating biotin-dynabead complex to 50C for 30 min but I don't think this works and that's why my yield is low.

As for why I'm getting only 10% membrane proteins, I still cannot figure it out.