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  1. Dec 2022
    1. Diagram of immunoprecipitation (IP) using either pre-immobilized or free antibodies.

      Immunoprecipitation is a technique for the isolation of protein or a complex (protein-protein interactions) Sample is combined with a specific antibody for the epitope of interest. The antibody-protein complex is removed and analysed.

      1. Molecules from biological sample (lysed) +incubated with antibodies (free or mounted onto support (like agarose bead, magnetic bead))
      2. protein A or G coupled beads added.
      3. Centrifuged
      4. Results in Beads with protein A/G bound to antibody-POI complex.
      5. Well separated in this way, differentially based on sedimentation coefficient.

      Co-immunoprecipitation (can isolate one type of protein in its complex)

      Isolate POI(s) Good with low conc. of POI Protein interactions Unknown proteins Determine if protein is actually being expressed in a given tissue.

      Western blot is carried out to analyse the output.

      Vary salt and detergent levels to preserve or destroy protein interactions.