EGFP sensitivity augmentation by anti-GFP Abs

Renee Lapointe lapointr at POST.QUEENSU.CA
Wed Oct 14 14:43:11 EST 1998


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?

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)?

Thank you for your help,

Renee Lapointe

Renée Lapointe
Department of Microbiology and Immunology
Queen's University, Kingston, Ontario, K7L 3N6
CANADA
Tel.:   (613) 545-6702, FAX: (613) 545-6796
E-mail: lapointr at post.queensu.ca





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