Interpreting an Affinity Chromatogram

The video tutorial explains the concept of chromatograms and their interpretation, focusing on affinity chromatography. It covers the principles of affinity chromatography, using examples with enzymes like hexokinase and lactate dehydrogenase. The tutorial details how proteins with specific affinities are separated using columns with analogs of substrates or coenzymes. It also discusses the analysis of chromatograms, identifying peaks, and the process of using wash and elution buffers to purify proteins. The video concludes with a discussion on the importance of affinity chromatography in protein purification.

What is a chromatogram and how is it used in affinity chromatography?

Explain the principle behind affinity chromatography.

How does the concept of affinity relate to the binding of proteins to substrates?

What factors can influence the elution order of proteins in affinity chromatography?

What is the difference between the non-binding region and the binding region in a chromatogram?

Explain how the structure of a protein affects its binding affinity in chromatography.

What role does the wash buffer play in the affinity chromatography process?

Describe the process of elution in affinity chromatography.

What is the significance of using a salting out technique in the elution buffer?

How do you determine which fractions contain the protein of interest after chromatography?