In an experiment you perform the following- you attach a flag epitope to HDAC, you over express a mutated Mad1-pro gene within the experimental cell, you immunoprecipitate the protein complex using flag epitope and run that on a non-denaturing SDS PAGE and western blot. Now you add antibodies for HDAC, MAD1 and SIN3A stripping after each addition. What will you observe?(a) Bands at same position every time(b) 3 bands at different positions one at a time(c) Bands at same position on 2 occasions(d) No bandI have been asked this question during an interview.I need to ask this question from Transcription in Eukaryotes : Chromatin Remodelling topic in section Gene Expression : Transcription Prokaryotes of Cytogenetics

In an experiment you perform the following- you attach a flag epitope

1.	In an experiment you perform the following- you attach a flag epitope to HDAC, you over express a mutated Mad1-pro gene within the experimental cell, you immunoprecipitate the protein complex using flag epitope and run that on a non-denaturing SDS PAGE and western blot. Now you add antibodies for HDAC, MAD1 and SIN3A stripping after each addition. What will you observe?(a) Bands at same position every time(b) 3 bands at different positions one at a time(c) Bands at same position on 2 occasions(d) No bandI have been asked this question during an interview.I need to ask this question from Transcription in Eukaryotes : Chromatin Remodelling topic in section Gene Expression : Transcription Prokaryotes of Cytogenetics
Answer» The CORRECT ANSWER is (c) BANDS at same position on 2 occasions Easy explanation: As Mad1-pro can’t bind to SIN3A due to the mutation it will not precipitate with the HDAC-SIN3A complex. Due to non-denaturing nature of the gel both the proteins MIGRATE together to the same distance so the band appears at the same position.

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