Absorbance at 480nm was recorded using a SpectraMax 250 microplate reader. We next investigated whether the various lysis buffers and luciferase reagents were compatible with the NanoOrange protein quantitation assay. For this assay system, fluorescence increases with increasing protein concentration, as seen in Figure 2 for the water control. At 1X concentrations, all lysis buffers and luciferase reagents did not appear to be compatible with this protein kit. Recommended Buffer Concentration for Performing Protein Quantitation.
Recommendations are for Promega lysis buffers and luciferase reagents as indicated.
For accurate protein quantitation using these or any other assay kits, the standard curve must be generated in the same dilution of lysis buffer or luciferase reagent. Then luciferase reagent is added to an equal volume of 1X PBS, which is added in turn to the cells. Do not add the reagent to standard culture media as sera will interfere with the protein assay. Alternatively, cells can be lysed using a lysis buffer, and an aliquot of the lysate is then used for protein determination as described above. Products may be covered by pending or issued patents.