recombinant protein - does GST affect the function? (Dec/26/2006 )
QUOTE (mdfenko @ Dec 27 2006, 04:08 PM)
QUOTE (Pumuki @ Dec 27 2006, 06:36 AM)
in the datasheet of the recombinant protein says that it can be used for these purposes: ELISA, WB, Antibody Production, Assay Development, Protein Array.
What does "assay development" mean? what kind of assays do they want to mean?
thanks
What does "assay development" mean? what kind of assays do they want to mean?
thanks
to "assay" is to determine how much of something is present. assay development is to devise a method to make such a determination.
the recombinant protein could be used: as a standard for an assay; as the displaced species in an immunodisplacement assay; if it is an enzyme then it could be used to find out how much of its substrate is present; if it is a substrate for an enzyme then it could be used to determine how much of that enzyme is present.
Oh, I understand it now
Many thanks mdfenko. -Pumuki-
Pumuki, i would definitely cleave off GST. You dont need it. it should be easy using thrombin like previuosly suggested. YOu can use the GST tagged protein to attach it to the beads for example and make some co-immunoprecipitation experiments with cell lysates. but if you dont need it, there is no need to leave it. unless you have problems with cleaving it off of course. 
-Kathy-