Bradford Protein Determination
1.) Use DNAse free Bovine serum albumine (1 mg/ ml in H2O from Pharmacia cat. 27-8915-01) and dilute 125 ul in 4.875 ml cold PBS. Mix well
2.) Prepare little 5-7 ml glass tubes and label 0 ug, 2.5 ug, 5 ug, 10 ug, 15 ug, 20 ug.
Then add in order 0.8 ml, 0.7 ml, 0.6 ml , 0.4 ml, 0.2 ml and 0 ml of cold PBS.
3.) Add in order from stock 0 ml, 0.1 ml, 0.2 ml, 0.4 ml, 0.6 ml, 0.8 ml of BSA. These are the standard tubes for the curve.
4.) Prepare three more tubes all with 0.8 ml PBS and add 1 ul, 5 ul and 10 ul of protein extract.
5.) Add to all tubes 200 ul Bradford Reagent mix (BIO Rad cat. 500-0006) and vortex well. Let it sit at RT for 5min.
6.) Read all tubes at visible light at 595 wave length.
7.) Plot numbers on a curve and protein concentration of sample.
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