Renee Lapointe wrote in message ...
>Hello everyone,
>I have two questions for the group.
>>1) I would like to get some information about the use of anti-GFP
>antibodies to augment the GFP signal in fluorescent microscopy. We have
>been using EGFP as a reporter for gene expression from various promoters,
>but it seems that EGFP needs to accumulate in the cells for a long time
>before the signal is detectable (24 hours). We are interested in knowing
>the % of cells which are expressing from these promoters regardless of the
>level of expression. Therefore we would like to know whether the use of a
>GFP specific antibody could improve the signal. Has anyone tested that?
>
Why not just use LacZ as reporter then?
>2) I guess this brings me to the second part of my message. CLONTECH, the
>company which sells these vectors, also sells a polyclonal antibody against
>GFP, but it's titer for immunofluorescence is 1/200, rendering the cost of
>its use too high for us to use (we have numerous promoters and conditions
>to test). Does anyone knows of a cheaper source of GFP-specific antibody
>which would detect EGFP even in a denatured form (for westerns for
example)?
>
X-gal staining for LacZ is very cheap.
Best regards,
Victor Leung
