Experiment 3

Question 1

Bradford Assay

Answer 1

• determine very small quantities of protein
• very rapid
• assay based on binding of CBB(coomassie brilliant blue)
• acidic condition when red (+1)=
• basic conditions when Blue (-1)-> can interact with basic residues of side chains

Question 2

Interactions between Dye and Protein

Answer 2

1. Ionic w/ side chains of basic residues
2. Hydrophobic interactions with aromatic rings
   Note* turns blue when binds to protein
   - intensity depends on amount of protein

Question 3

Ligands

Answer 3

Molecules reversible bind to protein

• ex hormones, drugs
• can determine y and kf
• experiment: CBB (ligand) binding to BSA

Question 4

Concentration of which protein is determined ?

Answer 4

Bovine serum albumin (BSA)