Protein Determinations Using Bradford Assay

Question 1

Bradford assays are rapid, relatively inexpensive and quite sensitive. T or F?

Answer 1

True

Question 2

Two common reducing agents used:

Answer 2

dithiothreitol and 2-mercaptoethanol

Question 3

Why can detergent not be used in the assay?

Answer 3

The detergent interferes with the binding of the Bradford reagent to the protein

Question 4

What is the dye reagent composed of?

Answer 4

dye solubilized in phosphoric acid and methanol

Question 5

What are the protein resides that Coomassie Brilliant Blue G-250 dye binds to?

Answer 5

arginine, tryptophan, tyrosine, histidine, and phenylalanine

Question 6

How many times as well does the dye bind to arginine as the other aromatic residues?

Answer 6

8 times

Question 7

What is the most frequently employed protein used to prepare standards?

Answer 7

Bovine serum albumin (BSA)

Question 8

Instead of the standard BSA, what is used in the Bradford assay?

Answer 8

Ovalbumin

Question 9

Is the dye in the solution in cationic or anionic form?

Answer 9

Cationic

Question 10

Is the bound dye in cationic or anionic form?

Answer 10

Anionic

Question 11

What is the spectrophotometer set at for the absorption of bound dye?

Answer 11

595 nm