4 replies to this topic

[CambridgeBiochemist]

Hi, I wish to double label each subunit of a homo tetramer protein. It can be done by introducing fluorescent artificial amino acids using in-vitro translation. But since that has failed to produce my protein, I am wondering whether there are any alternative ways...

Thank you in advance.

[madrius1]

Have you considered fusing your protein with fluorescent tags such as GFP, YFP, SFP, DS-Red, and so on?

Also, why do you want to label each subunit with 2 different fluorescent tags? Knowing that will greatly help on answers ^^

[CambridgeBiochemist]

madrius1, on Jan 11 2010, 08:47 PM, said:

Have you considered fusing your protein with fluorescent tags such as GFP, YFP, SFP, DS-Red, and so on?

Also, why do you want to label each subunit with 2 different fluorescent tags? Knowing that will greatly help on answers ^^

It's for FRET measurement to study conformational changes

[madrius1]

Can't help you then. Good luck!

[leelaram]

hi,
if you want to doubly label your protein with two different fluorophores you can introduce cysteines at the required position and label with alexa maleimide and TAMRA. i can send you the related articles if required

all the best