EGFP sensitivity augmentation by anti-GFP Abs

victor vicleung at hknet.com
Mon Oct 19 10:43:15 EST 1998


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





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