Jeremys Content Flashcards
What is a PathHunter assay?
A cell-based assay used to measure G protein-coupled receptor (GPCR) activation by detecting β-arrestin recruitment.
What are the two key components of a PathHunter assay?
A GPCR tagged with ProLink (PK, small fragment) and β-arrestin fused to Enzyme Acceptor (EA, large fragment)
How does the PathHunter assay detect GPCR activation?
By measuring the recruitment of β-arrestin, which brings PK and EA into proximity, forming a functional enzyme.
What is the role of β-galactosidase in the PathHunter assay?
t is the enzyme formed when PK and EA complement each other, hydrolyzing a substrate to generate a colorful fluorescence signal
What is the first step in a sandwich ELISA?
Coating the plate with a capture antibody specific to the target antigen.
What happens in the second step of a sandwich ELISA?
The sample containing the antigen is added and binds to the capture antibody.
What is added in the third step of a sandwich ELISA?
A detection antibody that binds to a different epitope of the antigen.
What is typically conjugated to the detection antibody in a sandwich ELISA?
An enzyme, such as horseradish peroxidase (HRP) or alkaline phosphatase.
What is the purpose of adding a substrate in the final step of a sandwich ELISA?
To produce a measurable signal (often colorimetric) when acted upon by the enzyme.
How does the signal intensity in a sandwich ELISA relate to the antigen concentration?
The signal intensity is directly proportional to the amount of antigen present in the sample.
What is a key advantage of sandwich ELISA over other ELISA formats?
It is highly specific and sensitive, capable of detecting antigens in complex mixtures.
What type of molecules are typically detected using sandwich ELISA?
Large molecules with multiple epitopes, such as proteins or peptides
Why was a natural and synthetic form of commendimide used?
It allows researchers to directly compare their properties and effectiveness, revealing subtle differences.
