Assay for quantification of protein by comparing measured Absorbance at 595 nm to bovine serum albumine standardBovine serum albumin (concentrated >200 \u00b5g\/ml)Roti-Quant 5x (ROTH)96 Well PlatePlate ReaderStandard measurementsDilute your BSA stock solution to 200\u00b5g\/ml in the same buffer used for your solution of interest.Dilute the Roti-Quant 5x reagent at a rate of 2:7,5. For 15 ml solution, this means 4 ml Roti-Quant added to 11 ml of Water.Create 200 \u00b5l each of standard concentrations according to the following table: ABC1Concentration [\u00b5g\/ml]Volume of stock neededVolume of solution buffer needed200200320201804404016056060140680801207100100100Work in Triplicates. Pipette 50\u00b5l of the standards into the 96-Well plateAdd 200 \u00b5l of the diluted Bradford reagent to each well. Incubate for 5 minutes at room temperatureMeasure the OD595 for each well. Create a regression line from the averages of each concentrations' measurementsProtein concentration measurementsDilute your solution of interest 1:20 or 1:40, depending on expected protein levels.Work in triplicates and use blanks. Pipette 50 \u00b5l of each dilution into a well on the 96-Well plate.When working with solutions containing pigments, prepare additional wells to which NO Bradford reagent will be addedAdd 200 \u00b5l of the diluted Bradford reagent to each well. For the pigment controls, add 200 \u00b5l of solution buffer instead. Optimally, use a multi-pipette to reduce the time needed for adding the reagent.Incubate for 5 minutes at room temperatureMeasure the OD595 for each well. When processing the data, correct the measurements for the blanks. When pigments are present in the solution, correct the OD595 by subtracting the OD595 of the pigment controls from the appropriate measurements.Determine the amount of protein by putting the achieved OD595 of your solution of interest into the regression formula and multiplying by the dilution factor.