You obtained the following raw data when setting up a Bradford standard curve: BSA (mg/ml) Absorbancy 595nm 0 0.225 1 0.310 2 0.420 3 0.510 4 0.610 5 0.720 6 0.810 7 0.915 8 0.950 9 0.980
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You obtained the following raw data when setting up a Bradford standard curve:
BSA (mg/ml) | Absorbancy 595nm |
0 | 0.225 |
1 | 0.310 |
2 | 0.420 |
3 | 0.510 |
4 | 0.610 |
5 | 0.720 |
6 | 0.810 |
7 | 0.915 |
8 | 0.950 |
9 | 0.980 |
10 | 0.990 |
After blanking against a bradford-dH2O sample, the protein concentration of an unknown sample was determined using the same method and an absorbancy of 0.570 was obtained.
Set up a standard curve, excluding outliers (experimental and statistical) and determine the protein concentration in the unknown sample in mg / ml (up to 3 significant figures).
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- You obtained the following raw data when setting up a Biuret standard curve: BSA (mg/ml) Absorbancy 540nm 0 0.158 1 0.210 2 0.260 3 0.305 4 0.360 5 0.410 6 0.455 7 0.510 8 0.530 9 0.550 10 0.554 After blanking against a biuret-dH2O sample, the protein concentration of an unknown sample was determined using the same method and an absorbancy of 0.262 was obtained. Set up a standard curve, excluding outliers (experimental and statistical) and determine the protein concentration in the unknown sample in mg / ml (up to 3 significant figures).Using the equation for the best-fit straight line through your data, the average absorbance of your unknown samples U1 and U2, and any dilution factors (DON’T LEAVE OUT THE DILUTION FACTOR), calculate the concentration of protein in the original unknown protein sample. y = 1.6849x + 0.0414R² = 0.9904Calculate protein concentration in unknown samples 1, 2, 3: Absorbance of Unknown 1 = 0.541 Absorbance of Unknown 2 = 0.85 Absorbance of Unknown 3 = 1.02 Standard Curve: Y = 0.0073x
- Using the data from the standard curve done previously, you record the absorbance of your extracted milk protein sample at 0.499 at 540nm. What is the mass of the protein in mg assuming you collected 10mL worth of the protein? Your standard curve should have a line of best fit with the equation y=0.0001x+0.0038.You record the absorbance of your unknown BSA sample at 0.276. What is the concentration in ug/mL of the unknown assuming you have a standard curve with the line of best fit with y=0.0001x+0.0038 as its equationConstruct a standard curve (absorbance versus protein concentration in the standards) Conc. (mg/ml) 0 0.25 0.5 0.75 1.0 Absorbance 0.000 0.126 0.249 0.361 0.464
- If you had measured the absorbances of your unknown in test tubes that were half the diameter of the ones that you used to make the standard curve, how would the mg of protein in the samples be affected? (Higher, Lower, No Change)When you plot the protein concentration against absorbance you expect to have a ______________ linear correlation because the ____________ the protein concentration, the ______________ absorbance.Table 2. Volume of BSA, protein content, and absorbance readings of reference solutions Solution Volume of BSA standard solution (μL) Protein content(μg/mL) Absorbance value At 595 nm 1 0 0 0 2 10 1 0.022 3 30 3 0.065 4 50 5 0.106 5 70 7 0.178 6 100 10 0.299 7 120 12 0.380 Make a graph by plotting the absorbance values versus the BSA protein content (in μg) for theseven reference solutions. When constructing the graph, be…
- The pH probe/meter uses following equations: Ecell = L + 0.0592 log a1 = L - 0.0592 pH Where L = L1 + EAg/AgCI + Easy= constants L1 = - 0.0592 log a2 a1 = activity of analyte solution a2 = activity of internal solution How will measured pH value be affected vs “real” pH if the temperature of the sample is 30C when pH was measured ? a.measured pH is lower than real pH b.impact can not be determined c.measured pH is higher than real pH d.measured pH is same as real pHIf I had a standard with 2.0 mL of Bradford reagent then added 50μL of 250 μL/mL ; what would be the concentration of original protein?Viscosity measurements were done on native form of Protein XYZ and on 4 denaturants. The crude Protein XYZ was diluted to produce 5% w/v. Using the Ostwald viscometer, the following data were obtained: time, sec time, sec time, sec time, sec Denaturant Blank (Native) Native Blank (Denaturant) Denatured pH= 2.00 70 95 70 101 Temp (90 °C) 70 98 70 150 0.1M Urea 72 97 88 110 DTT 71 96 90 170 a. Calculate and tabulate the reduced viscosities (in mL/g) of the native and denatured Protein XYZ. b. Determine two most effective denaturants. c. What can be inferred with the two most effective denaturants? Dont reject if you dont know the answer!