Question

Describe these two techniques: co-immunoprecipitation and sandwich ELISA. For each technique, be sure to explain its purpose and describe the main steps involved in carrying out the technique. (please answer everything)

Answer 1

CO-IMMUNOPRECIPITATION :-

Purpose :- it is a technique for the identification of the protein - protein interaction . It capturr the protein that are bound to specific target protein.

STEPS :-

1.Transfer cell with protein of interest .

2.Prepare cell lysate with proper lysis buffer.

3.Add antibody and resin.

4.Precipitate antibody conjugated protein complex.

5.Analyze by immunoblotting .

SANDWICH ELISA :-

Purpose :- it is a technique use to quantify the antigen between two layers of antiantibody.

STEPS :-

1.well coated with capture antibody

2. Non specific binding site blocked and antigen added.

3. Detection antibody added

4. Enzyme conjugated detection reagent added.

5. Substrate added and plate read at 420 nm.