I've a problem , i did an overexpression with fusion protein containning a transcription factor with gfp-G4
I did my transfection on HEK cells with lipofectamine. Once with GFP alone ( from another compagny) and the one from (Biorbyt) I checked after 24 hours of transcription and i saw in each plate the green signal that means the transfection works.
But then i did my western blot . I used the antibody GFP roche, cat. no. 11 814 460 001 and i didn't find any signal for it on my extract. Then my boss said your transfection does't work, but 2 of us saw the green signal by microscope.
I'm thinking maybe this antibody can't reconize the GFP, when it's associated (fusion) with another factor
Do you have any idea m to solve this problem.