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anti-FLAG (M2) cross reaction - (Sep/01/2009 )

Hi,
Does anyoone know if the anti-FLAG antibody M2 cross reacts with GFP?
Thanks!

-PiDC-

PiDC on Sep 1 2009, 10:44 AM said:

Hi,
Does anyoone know if the anti-FLAG antibody M2 cross reacts with GFP?
Thanks!


I never saw any cross reactivity with the Sigma F1804 M2 with turbo GFP, not sure about GFP or eGFP.

-Dr Teeth-

Dr Teeth on Sep 1 2009, 05:12 PM said:

PiDC on Sep 1 2009, 10:44 AM said:

Hi,
Does anyoone know if the anti-FLAG antibody M2 cross reacts with GFP?
Thanks!


I never saw any cross reactivity with the Sigma F1804 M2 with turbo GFP, not sure about GFP or eGFP.



I don't know how different is turboGFP from eGFP... I am using eGFP, which comes from Aequorea victoria, and the M2 antibody gives a band, in my hands, that correspond to the size of GFP (in a lane loaded whith GFP).
Thanks for your help!

-PiDC-

PiDC on Sep 1 2009, 11:31 AM said:

Dr Teeth on Sep 1 2009, 05:12 PM said:

PiDC on Sep 1 2009, 10:44 AM said:

Hi,
Does anyoone know if the anti-FLAG antibody M2 cross reacts with GFP?
Thanks!


I never saw any cross reactivity with the Sigma F1804 M2 with turbo GFP, not sure about GFP or eGFP.



I don't know how different is turboGFP from eGFP... I am using eGFP, which comes from Aequorea victoria, and the M2 antibody gives a band, in my hands, that correspond to the size of GFP (in a lane loaded whith GFP).
Thanks for your help!


I suppose given the 8 amino acid epitope for FLAG, it is likely that it may cross react with many proteins.
FYI, eGFP is not from Aequorea victoria. It is a F64L point mutant of the wild-type GFP, which is from the jellyfish, resulting in an "enhanced" GFP that is brighter, and I believe also more yellow. Good luck with the Westerns!

-Dr Teeth-

Have you tried blasting the flag tag sequence against the gfp sequence?

-miBunny-

Thank you very much!! :P

-PiDC-

Yes, Ive seen it quite often. Flag antibody reacts with gfp and the other way around. If you have a big amount of Flag-protein your anti gfp antibody will recognize the flag-protein

-laurequillo-