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Double Labelling of Same Protein


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#1 CambridgeBiochemist

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Posted 11 January 2010 - 04:45 AM

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.

#2 madrius1

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Posted 11 January 2010 - 12:47 PM

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 ^^

#3 CambridgeBiochemist

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Posted 12 January 2010 - 02:07 AM

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

#4 madrius1

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Posted 12 January 2010 - 10:26 AM

Can't help you then. Good luck!

#5 leelaram

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Posted 07 May 2011 - 08:49 PM

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
M. Naga Leelaram,
PhD student,
Indian Institute of Science,
Bangalore
India




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