GFP sandwich ELISA with immobilized GFP-A1(Cys)


Overview
Abstract
This application involves the identification of antigens in solution by sandwich ELISA using nanoCLAMPs covalently conjugated to maleimide coated microtiter wells via their lone C-terminal Cys. Antigen bound by the immobilized nanoCLAMP is identified by an enzyme-conjugated secondary antibody followed by development with the appropriate secondary substrate.
Type
  • ELISA
Target
Scores
Outcome
Successful
Signal
Acceptable
Cross Reactivity
Acceptable
Background
Acceptable
Dynamic Range
Acceptable
Results
Figure

Image 

GFP ELISA with capture by nanoCLAMP GFP-A1

Legend 

Analysis of the interaction between GFP-A1(Cys) and GFP by sandwich ELISA.  GFP-A1(Cys) was conjugated to maleimide coated microtiter wells of 96 well strips via the lone C-terminal Cys, at 5 µg/ml. Dilutions of GFP-V5 were incubated on the strips, washed, and then incubated with anti-V5-HRP and developed with TMB Ultra. Reactions were stopped with H2S04 and absorbance measured at A450. The blank wells were processed the same, except they included no GFP-A1(Cys).