Bradford - Protein Determination
Dr. Mario Lebendiker

Protein Purification Unit
 
 

Reagents & Solution
 
 

Bovine Serum Albumine (BSA) 10X: 1mg/ml H2O. Keep -20C in aliquots.

BSA 1X (0.1mg/ml): dilute BSA 10X in the buffer of your sample.

Bradford solution from Bio Rad Protein Assay. Kep in dark at 4C. (Do not use directly from the bottle. Transfer the volume you are going to use to another recipient).
 
 

Mini:for 500ul determination in Eppendorf tubes.

 
 
BSA 100ug/ml (ul)
H2O (ul)
Bradford (ul)
 total ug BSA
0
400
100
0
10
390
100
1
20
380
100
2
35
375
100
3.5
50
350
100
5
X ul sample
complete 400
100
X ug

Mix and read between 2 and 60 min at OD590nm. Plot OD590nm vs. ug BSA.

Try to use a sample volume with OD590nm near the middle of the curve.

Prepare triplicates of your sample, or use different points.
 
 

Elisa plates:for 125ul determination in 96 microwell plates.

 
BSA 100ug/ml (ul)
H2O (ul)
Bradford (ul)
 total ug BSA
0
100
25
0
3
97
25
0.3
6
94
25
0.6
9
91
25
0.9
12
88
25
1.2
X ul sample
complete 100
25
X ug

Mix and read immediately at OD590nm. Plot OD590nm vs. ug BSA.

Try to use a sample volume with OD590nm near the middle of the curve.

Prepare triplicates of your sample, or use different points.