Biotechniques 373 Pre Mid term Notes Review

Study Gradually

In A Sephadex g-75 Column, It has an Exclusion limit of 3 and 80 KdA If a molecule is 90kd and 100 Kda what will happen when it runs through the column

Both Molecules that are over the exclusion limit of 80 KDa will elute together at the beginning of separation

Which Buffer choice could you use while preforming an ion exchange chromatography with a cation exchanger, Such as Carboxymethyl (PKA:3.5)?

At pH 5 would be the Ideal Buffer pH since the PH would be larger than the pH/pka giving us a Negative charge that would be opposite, charged and attracted to the cation exchanger.

What is the Total Volume in the GFC?

(VT)- is the total Volume of the fraction where the Separation ends.

Which if the Following is a Cation exchanger ?

Which of the following proteins would cause a higher absorbance with Coomassie blue?

How does ninhydrin work?

It binds to the Amino group of Amino Acids

You have isolated some GFP from Bacteria. Which of the following tells you about the purest sample?

Known as the Highest ratio between the Total Amount of GFP over the Total Proteins.

What is Responsible For the Florescence of GFP?

A Sequence of 3 Amino Acids.

How does Coomassie Brilliant Blue G-250 make proteins quantifiable after binding them??

it switches the Protein sample absorbance from 465 nm to 595 nm

When making a buffer by titration, you can start only from the weak base

False

Using BSA as Bradford standard is an issue with pure Proteins, not with protein mixes.

True. Since BSA is a purified protein and has a high concentration of basic amino acids can change the Outcome of adding A Bradford assay to the protein solution.

The Kd Value of a Molecule is Always the Same.

True As the resin carries a Certain Exclusion limit the KD( Distribution coefficient will be the same).