What is the purpose of RIA?

RIA uses radiolabeled antibodies. The presence of target proteins

or antibodies is determined by measuring the amount go
radioactivity

What is the common use of RIA?
RIA's are more excessively used in the medical field to measure
relative amounts of hormones or drugs in patients sera.

What is the basic protocol of RIA?
1. A known amount of radiolabeled antigen is incubated with a
known amount of an antibody specific to the target antigen
2. antibody complexes are isolated
3. total amount of radioactivity is measured

What is the process of using RIA to analyze insulin?
1. insulin synthesized with a known amount of I-labeled
tyrosine's is incubated with a known amount of antibody specific
to antigen
2. insulin antibody complexes are isolated
3. total radioactivity amount is measured
4. unlabeled insulin (cold insulin) mixed into solution in
increased amounts. the cold insulin competes with labeled
insulin (hot insulin) for the antibody. As more cold insulin is
added less radioactivity is measured and recovered. (competition
determines standard curve)
5. repeat steps 1-3 using patients serum instead of cold insulin

What is the purpose of the standard curve in RIA?
To estimate the amount of insulin circulating in a patients serum