3 replies to this topic

[utopia]

hey folks,
can anybody help me please, which one of these methods is the most efficient one to showing an ínteraction bwn 2 proteins?

*Yeast 2-hybrid-system
*affinity chromatography
*co-immunopricipation
* ??or do you have a better idea?

Immunohistochemically, we could show the colocalization between 2 membranproteins. The question is now: do these proteins interact with each other directly and therefore we get a colocalisationsignal or they are just in close neighbourhood of each other and due to this adjacency we get a signal.

which method have I to use in order to be able to prove the colocalisation?

thank you in advance
Shokufeh

[jadefalcon]

Far-Westernblotting, perhaps?

mike

[Simonsays]

Hi
If you have the necessary antibodies to immunoprecipitate and blot, I would strongly suggest co-immunoprecipitation.

Simon

[TheMadScientist]

Does anyone know of any techniques to crosslink associated proteins for subsequent co-iping? I seem to remember a UV-crosslinking chemical for protein-protein interactions, separate from a DNA-protein UV crosslinking assay. Thanks ahead of time!