2 replies to this topic

[sebwood]

Hi

I just received cells tranfected with receptor-GFP.
I can't see GFP by direct microscopy. The person who send me cells see a great GFP signal...

But by Western and IF I can say that GFP is in my cells ( and also the receptor ).

She send cells twice but it's always the same problem.


Anyone already have the same probl ?

Thanks for reply

[Simonsays]

That may sound a little too simple, but do you watch your cells in the dark?? GFP needs to be excited so it can emit some green light.
Excitation wavelength : 488nm
Emission wavelength : 509nm

The light from a room is enough, generally.

Simon

[sebwood]

hi

of course i protect the cells from light during fixation etc......

and it's not a problem of wavelenght, I work often on confocal.....

thx 4 ur reply