Question

In: Biology

You have a protein, which has high affinity to DNA. You prepared resin attached with a...

You have a protein, which has high affinity to DNA. You prepared resin attached with a small piece of DNA. After allowing the protein to bind to the resin, how would you elute the protein?

Solutions

Expert Solution

You have a protein, which has high affinity to DNA. You prepared resin attached with a small piece of DNA. After allowing the protein to bind to the resin, you can elute the protein by affinity column chromatography.

Affinity chromatography relies on the specific and reversible binding of a protein to an immobilized ligand. The ligand can bind directly to either the protein of interest or a tag that is covalently attached to the protein.

Affinity column chromatography involves the following steps:

  1. Incubate crude sample with the affinity support to allow the target molecule in the sample to bind to the immobilized ligand.
  2. Wash away non-bound sample components from the support using appropriate buffers that maintain the binding interaction between target and ligand.
  3. Dissociate and recover the target molecule by elution from the immobilized ligand by altering the buffer conditions so that the binding interaction no longer occurs.

Ps- An upvote would be highly appreciated


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