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Cellular and Molecular Biology

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Immunoprecipitation of Specific Proteins

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You are measuring the amount of Protein AAA, a protein involved in neurodegenerative diseases, in the cytoplasm of a neuronal culture by immunoprecipitation and gel electrophoresis.

Following lysis of the neurons and incubation with the antisera, you immunoprecipitate the protein and analyze it by separation on an SDS page gel followed by western blotting. Your western blot reveals that there are not one but three different protein bands.

Which of the following could account for this finding? Select ALL that apply.

A

The neurons contained several different isoforms of the protein, all with the same antigenic site.

B

Your primary antibody cross-reacted with other proteins in the neurons.

C

The protein is interacting with itself to form aggregates of several sizes.

D

The primary antibody was contaminated with other antibodies.

E

The protein was degraded in the procedure and the antibody is recognizing multiple fragments.