GFP to expression - (Nov/29/2006 )
We have to bring GFP to expression with 2 methods, 1 of those must be immonulogic.
We don't exactly know how to do this. Western blotting, ELISA?
And than the second method?
We will get an plasmid with the GFPgene inside, the plasmid is pGEX261 and has a ptac-promotor.
Could you help us?
-Coebergh-
QUOTE (Coebergh @ Nov 29 2006, 06:34 PM)
We have to bring GFP to expression with 2 methods, 1 of those must be immonulogic.
We don't exactly know how to do this. Western blotting, ELISA?
And than the second method?
We will get an plasmid with the GFPgene inside, the plasmid is pGEX261 and has a ptac-promotor.
Could you help us?
We don't exactly know how to do this. Western blotting, ELISA?
And than the second method?
We will get an plasmid with the GFPgene inside, the plasmid is pGEX261 and has a ptac-promotor.
Could you help us?
Wb is a good choice, various companies offer anti-GFP Ab
-The Bearer-
We have chosen ELISA. What do we need for this?
And does GST have any influence on the experiment with LB/amp? Do we have to put something extra in it because of the GSTgen in the plasmid?
-Coebergh-
QUOTE (Coebergh @ Dec 1 2006, 11:53 AM)
We have chosen ELISA. What do we need for this?
And does GST have any influence on the experiment with LB/amp? Do we have to put something extra in it because of the GSTgen in the plasmid?
And does GST have any influence on the experiment with LB/amp? Do we have to put something extra in it because of the GSTgen in the plasmid?
if you try ELISA you need a la long more Ab than for Wb; anyway, if you prefer to measure in a microplate system, you may perform instead of ELISA direct measurement at ~488 nm (+/- 20 nm tolerable) of GFP; you need microtiterplates for fluorescence measurement and cell culture f.i. Cytowell (Nunc), and a plate reader for fluorescence; ELISA does only make sense if you have no such equipment...
-The Bearer-