Advanced Protein Assay Methods

Advanced Protein Assay: 5x Stock Methods (Cat. # ADV01)

Method 1 for concentrated protein solutions (>3.0 mg/ml):

1. Pipette 5 ml of 1X Advanced Protein Assay Reagent (Cat. # ADV01) into a 10 ml disposable glass tube.

2. Pipette 5 µl of protein solution into the tube containing 5 ml ADV01 and vortex 5 s.

3. Blank the spectrophotometer on 1X ADV01 and read absorbance of your sample at between 570 and 615 nm (optimal 590 nm).

4. Calculate protein concentration based on 1.0 OD_{570 to 615 nm}= 30 µg protein per ml reagent per cm.

Method 2 for dilute protein solutions (<3.0 mg/ml):

1. Pipette 1 ml of 1X Advanced Protein Assay Reagent (Cat. # ADV01) into a 1.5 ml disposable microfuge tube.

2. Pipette 10 µl of protein solution into the tube containing 1.0 ml ADV01.

3. Blank the spectrophotometer on 1X ADV01 and read absorbance of your sample at between 570 and 615 nm (optimal 590 nm).

4. Calculate protein concentration based on 1.0 OD_{570 to 615 nm}= 30 µg protein per ml reagent per cm.

Method 3 for very dilute protein solutions (<0.1 mg/ml):

1. Pipette 0.2 ml of 5X ADV01 into two 1.5 ml disposable microfuge tubes and label A and B.

2. Pipette 0.8 ml of protein solution into the tube A.

3. Pipette 0.8 ml of protein solution buffer only into the tube B.

4. Blank the spectrophotometer on contents of tube B, and read absorbance of your sample at between 570 and 615 nm (optimal 590 nm).

5. Calculate protein concentration based on 1.0 OD_{570 to 615 nm}= 30 µg protein per ml reagent per cm.

Method 4 for concentrated protein solutions (>3 mg/ml) in 96-well format:

1. Pipette 3.0 µl of protein solution into two wells A1 and A2.

2. Pipette 0.3 ml of 1X ADV01 into wells A1, A2, B1 and B2.

3. Place in reader and measure absorbance at 570 to 615 nm, optimally 590 nm.

4. Calculate protein concentration based on 1.0 OD_{570 to 615 nm}= 37.5 µg protein per ml reagent per 0.8 cm.

Method 5 for dilute protein solutions (<3 mg/ml) in 96-well format:

1. Pipette 10 µl of protein solution into two wells A1 and A2.

2. Pipette 0.3 ml of 1X ADV01 into wells A1, A2, B1 and B2.

3. Place in reader and measure absorbance at 570 to 615 nm, optimally 590 nm.

4. Calculate protein concentration based on 1.0 OD_{570 to 615 nm}= 37.5 µg protein per ml reagent per 0.8 cm.