A Non - Interfering™ ( NI ) Protein Assay
Perform assays at room temperature .
Use 2ml tubes for assay .
Prepare a set of protein standards using the supplied BSA or Non - Animal Protein Standard as indicated in the table below :
Tube # 123456Protein Standard [ 2mg / ml ] ( µl ) 048122025Protein ( µg ) 0816244050 .
Add 1 - 50µl of the protein samples to be assayed to 2ml tubes .
Add 0.5ml UPPA™ I to each tube and vortex .
Incubate for 2 - 3 minutes at room temperature .
Add 0.5ml UPPA™ II to the tubes and vortex .
Centrifuge the tubes at maximum speed ( ~ 10,000xg ) for 5 minutes to pellet the precipitated protein .
Decant off the supernatant , return the tubes to the centrifuge as before , quickly pulse to spin down residual liquid and remove with a pipette .
Add 100µl Copper Solution ( Reagent I ) and 400µl deionized water to the tubes and vortex until the protein precipitate pellet dissolves .
Using 1ml pipette , rapidly shoot 1ml Reagent II directly into each tube containing Reagent I plus DI Water and immediately mix it by inverting the tubes .
Incubate at room temperature for 15 - 20 minutes and then immediately read absorbances at 480nm against DI water .
Plot absorbance against protein concentration and determine protein concentrations of unknowns .
