A Non - Interfering™ ( NI ) Protein Assay ( high throughput 96 - well )
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 .
Add 0.5ml UPPA™ II to the tubes and vortex .
Incubate for 2 - 3 minutes at room temperature .
Centrifuge the titer plate at ~ 5,000xg for 7 minutes to pellet the precipitate .
Invert the titer plate to remove the supernatant and shake to remove all excess supernatant .
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 .
After incubation , transfer 200µl assay reaction to a flat bottom 96 well micro titer plate and measure the absorbances at 480nm against DI water .
Plot absorbance against protein concentration and determine protein concentrations of unknowns .
