In the preparation of the standard curve for protein analysis, 50 mg BSA (bovine serum albumin) dissolved in water to a final volume of 5 mL with a stock solution. Based upon the dilutions of each standard a standard curve was created as demonstrated by Figure 1. Two common proteins used for standard curves are bovine serum albumin (BSA) and an immunoglobin (IgG). Use the obtained value of slope (a) to calculate protein concentration in samples. Prepare 50 ml of 1 mg/ml stock solution of a standard protein (the most commonly used protein is bovine serum albumin or BSA) in distilled water. To create the standard curve, I have measured the absorbance of 8 standards (25, 125, 250, 500, 750, 1000, 1500 and 2000 μg/mL total protein) and a blank sample (0 μg/mL total protein) for background corrections. To calculate the concentration of the undiluted, unknown sample, simply multiply by the dilution factor. So, 0.5 x 10= 5mg/ml. Protein Standard Curve or Linear Plot. What is the weight of BSA in (a) 0.1 of Blank (0 Standard, #8) from all readings. Prepare bovine serum albumin (BSA) standard: Dissolve 0.05 g BSA in 5 mL PBS to obtain 10 mg/mL BSA standard. You need to make standard curve drawing with Bradford Method. Using the measurement of 595nm, the standard curve was plotted. Perform the assay and calculate the standard (see below). The result should be around 0.5mg/ml. Prepare Coomassie Brilliant Blue G-250 staining solution: Dissolve 50 mg Coomassie Brilliant Blue G-250 in 25 mL 90% ethanol, add 50 mL phosphoric acid (85%) , and dilute with pure … Here is the data: What we have here is the average absorbances of each standard next to the corresponding known concentrations. From the working solution, prepare 10 different solutions with the concentration ranging from 10 µg to 100 µg and maintain the total volume of 100 µl with distilled water. Plot the Standard curve, OD 562 (on Y-axis) vs Standard BSA concentration (on X-axis). Based on calculations a y value of 0.000x + 0.153 was obtained and a regression line (R 2) value of 0.902. BSA standard calibration curve. Here are the details. Obtain the equation from the plot Y = aX + b. These two proteins have different amino acid compositions, which lead to a different standard curve and a slight difference in the final determination of the unknown protein concentration.