Protein Concentration Determination

Question 1

Techniques for Protein Concentration Determination

Answer 1

1. Bradford Assay : 595 nm
2. Bicinchoninic Acid (BCA) Assay : 562 nm
3. A280 : 280 nm

Question 2

Bradford Assay

Answer 2

+
Not affected for reducing agents

• Not compatible w/ detergents or basic conditions
  Depened on sequence of protein

Question 3

Bicinchoninic Acid (BCA) Assay

Answer 3

+
Not sensitive detergents
Protein backbone

-
Not compatible with reducing agents or anything that chelates copper
Standard curve generated from a standard protein

Question 4

A280

Answer 4

+
Sample is recoverable
Simple
Can be perfomed in native or denaturing buffer

-
Require pure samples
Not compatible w/ nucleic acids
Depends on sequence of your protein