Alternative methods to detect protein:protein interaction
Posted 06 June 2009 - 07:42 PM
I have some enquiries:
- I use In vitro translation to express the flag-prey and bait proteins "labeled with 35S-met" to test interactions.
then I expose on phosphor screen and scan using Typhoon machind, but the problem is our machine down for long time.
Could you help me find alternative methods to detect interactions where i deal with mammalian proteins.
I need sensitive methods .
Posted 07 June 2009 - 01:06 PM
- Far Western (blot protein A on membrane, use tagged protein B as first antibody, use tag-antibody as second antibody (eg HIS))
- Co-Purification (bind tagged protein A on a column, put protein B on the column, wash and elute and see wether protein B is washed out or elutes together with protein B )
- Native PAGE (incubate protein A with protein B and both proteins alone, put the samples on a Native Gel, see wether complexes are formed between A and B )
Edited by mastermi, 07 June 2009 - 01:07 PM.
Posted 09 June 2009 - 05:51 PM
I mean also alternative methods to detect radioactive proteins.
Posted 10 June 2009 - 06:55 AM
genius does what it must
i do what i get paid to do