A Bradford assay was carried out to measure protein concentration. The absorbance of five standard samples...
A Bradford assay was carried out to measure protein concentration. The absorbance of five standard samples and two unknown samples are listed in the table below. The Bradford reaction mixture (1 ml) was prepared as follows: 100 μl sample A or sample B + 200 μl Bradford reagent + 700μl 0.9% NaCl solution
| sample ID | concentration (mM) | absorbance (O.D.) |
| std 1 | 100 | 0.78 |
|
std 2 |
50 | 0.4 |
| std 3 | 25 | 0.18 |
| std 4 | 12.5 | 0.08 |
| std 5 | 0 | 0 |
| unknown sample A | unknown | 0.56 |
| unknown sample B | unknown | 0.11 |
Make a standard curve using the absorbance (O.D.) as the x axis and the concentration (mM) as the y axis. Find the equation of the standard curve.
hint: (1) follow the instruction regarding which number should be on the x or y axis; (2) unknown sample A and B were diluted in the process of preparing the Bradford reaction. To figure out the original concentration in Q3 and Q4, you need to figure out the concentration and the volume of the diluted solution first ( = bradford reaction).
This question was already on chegg but the mm were wrong for a and b. please read the hint.
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A Bradford assay was carried out to measure protein
concentration. The absorbance of five standard samples...
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs...
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs samples of known BSA concentrations. Now you need to determine the amount of protein in two mitochondria samples taken from fish, one taken from fish under normal conditions and the other taken from fish under oxidative stress. The absorbance reading for the normal fish fraction was A595=175, and the absorbance reading for the stressed fish fraction was A595= 275 Bradford Assay Standard Curve y...
In biochemistry, we use a 'Bradford Assay' to calculate the concentration of protein in a sample....
In biochemistry, we use a 'Bradford Assay' to calculate the concentration of protein in a sample. This method uses the same principles of the standard curve from this experiment. In a Bradford Assay, a standard curve is prepared using a known protein, typically Bovine Serum Albumin (BSA), but rather than measuring the density, we can measure a different property - its 'absorbance'. The graph below is representative of a standard curve from a Bradford Assay (with the line equation displayed...
I forgot to put down that the observance (=OD) at 280nm is 0.679A Experiment 8: Bradford...
I forgot to put down that the observance (=OD) at 280nm is
0.679A
Experiment 8: Bradford assay and UV-method for determination of protein concentrations Materials - An ice bucket with ice, your purified sample, three visible plastic cuvettes for Bradford assay, one UV plastic cuvette for UV-method, a piece of parafilm, a Vis-spec, a Lab Quest, Bradford reagent, white tape (share), a sharpie. Bradford assay (reference 21) 1. Add 990 uk Bradford assay reagent to a plastic cuvette 2. Blank...
A Bill' external standard curve built from absorbance signal vs. analyte concentration (in uM) has the...
A Bill' external standard curve built from absorbance signal vs. analyte concentration (in uM) has the following calibration curve equation y = 0.0500 x + 0.003. If a sample produced an absorbance of 0.300, what was the Bi concentration in the solution in the cuvette? A. 0.015 mM B. 6.00 mm c. 5.94 mm D. 0.600 mm Reset Selection estion 4 of 5 2 Points If the concentration of Bi in a diluted sample (after dilution of the original sample)...
In lab, you diluted your unknown protein solution by combining 14 ?L of unknown protein solution...
In lab, you diluted your unknown protein solution by combining
14 ?L of unknown protein solution with enough
PBS buffer to obtain a total volume of 100 ?L, then added 1000 ?L Bradford reagent. After waiting ten
minutes, the absorbance at 595 nm was 0.206. Using the standard
curve from the previous question, calculate the total concentration
of protein (in ?gmL) in the original sample.
Slope = 0.02371 Intercept = 0.01132 r2
= 0.973
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the...
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the Part II Data Table and the equation on page 2 for the absorbance assay at 280 nm. Hint: Convert the molar concentration (M or moles/liter) of BSA from absorbance assay at 280 nm method to mg/mL, assume that the molecular weight of BSA = 66.5 kDa [7 points 6500 g/mol Post-lab 10 Report Form: Determination the Concentration of a Protein You must show your...
Question 14 4 pts A protein assay was performed using the Bradford Reagent to determine the...
Question 14 4 pts A protein assay was performed using the Bradford Reagent to determine the protein concentrations of a series of pet food extract samples. The BSA Standard Curve had a slope of 0.0173 Data obtained for the pet food are shown in the table below. Tubes 1-2 contained different volumes of the pet food extract. Tube # Volume Sample (UL) A(595) Volume Volume Water Reagent (3.0 (L) mL) 803 503 1 2 20 50 0.293 0.657 What is...
Calibration Curve Standard Number [Fe mg/ml Absorbance 0.001 0.228 2 0.002 0412 0.003 s. 622 0.004...
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Answer 2) To measure the concentration of phosphorous in a water sample, Phosphorous was first converted...
Answer
2) To measure the concentration of phosphorous in a water sample, Phosphorous was first converted into a color complex in the presence of ammonium molybdate and SnClo. The absorbance of a series of standard phosphorous solutions was measured by a spectrophotometer and gave the calibration curve y 1.150 x, where y is the absorbance of phosphorous complex at 690 nm while x is the concentration of P in ppm. An unknown natural water sample was collected. 10.00 mL of...
Data Results (show all work on following pages) Standard Dilutions A-E Solution Absorbance А B Concentration(mg/L)...
Data Results (show all work on following pages) Standard Dilutions A-E Solution Absorbance А B Concentration(mg/L) 0.005 0.001 0.002 0-003 0-004 C 1. 188 0.232 0.47 0.702 0.953 D E Graph of Absorbance versus Concentration (mg/L) 81 Show work for concentration values Winai is in equation for mine samaru curve from the repuri dain? Extraction of B-carotene Mass or carrot 5.49 Absorbance of carrot solution (diluted if 0.455 necessary) Concentration of carrot solution (diluted 0.00194 if necessary)(mg/L) Concentration of original...
1. You have generated the following standard curve after a Bradford Assay to measure Absorbance vs samples of known BSA concentrations. Now you need to determine the amount of protein in two mitochondria samples taken from fish, one taken from fish under normal conditions and the other taken from fish under oxidative stress. The absorbance reading for the normal fish fraction was A595=175, and the absorbance reading for the stressed fish fraction was A595= 275 Bradford Assay Standard Curve y...
In biochemistry, we use a 'Bradford Assay' to calculate the concentration of protein in a sample. This method uses the same principles of the standard curve from this experiment. In a Bradford Assay, a standard curve is prepared using a known protein, typically Bovine Serum Albumin (BSA), but rather than measuring the density, we can measure a different property - its 'absorbance'. The graph below is representative of a standard curve from a Bradford Assay (with the line equation displayed...
I forgot to put down that the observance (=OD) at 280nm is
0.679A
Experiment 8: Bradford assay and UV-method for determination of protein concentrations Materials - An ice bucket with ice, your purified sample, three visible plastic cuvettes for Bradford assay, one UV plastic cuvette for UV-method, a piece of parafilm, a Vis-spec, a Lab Quest, Bradford reagent, white tape (share), a sharpie. Bradford assay (reference 21) 1. Add 990 uk Bradford assay reagent to a plastic cuvette 2. Blank...
A Bill' external standard curve built from absorbance signal vs. analyte concentration (in uM) has the following calibration curve equation y = 0.0500 x + 0.003. If a sample produced an absorbance of 0.300, what was the Bi concentration in the solution in the cuvette? A. 0.015 mM B. 6.00 mm c. 5.94 mm D. 0.600 mm Reset Selection estion 4 of 5 2 Points If the concentration of Bi in a diluted sample (after dilution of the original sample)...
In lab, you diluted your unknown protein solution by combining
14 ?L of unknown protein solution with enough
PBS buffer to obtain a total volume of 100 ?L, then added 1000 ?L Bradford reagent. After waiting ten
minutes, the absorbance at 595 nm was 0.206. Using the standard
curve from the previous question, calculate the total concentration
of protein (in ?gmL) in the original sample.
Slope = 0.02371 Intercept = 0.01132 r2
= 0.973
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the Part II Data Table and the equation on page 2 for the absorbance assay at 280 nm. Hint: Convert the molar concentration (M or moles/liter) of BSA from absorbance assay at 280 nm method to mg/mL, assume that the molecular weight of BSA = 66.5 kDa [7 points 6500 g/mol Post-lab 10 Report Form: Determination the Concentration of a Protein You must show your...
Question 14 4 pts A protein assay was performed using the Bradford Reagent to determine the protein concentrations of a series of pet food extract samples. The BSA Standard Curve had a slope of 0.0173 Data obtained for the pet food are shown in the table below. Tubes 1-2 contained different volumes of the pet food extract. Tube # Volume Sample (UL) A(595) Volume Volume Water Reagent (3.0 (L) mL) 803 503 1 2 20 50 0.293 0.657 What is...
Calibration Curve Standard Number [Fe mg/ml Absorbance 0.001 0.228 2 0.002 0412 0.003 s. 622 0.004 0802 esos 0.990 Show example calculation for Fe concentration of standard number 1: 5 L Use Excel to construct the calibration curve. Include the regression line and equation. Submit with your report Mass of unknown Fe sample_0.1026 Sample unknown mass diluted to 50 mL Trial Sample solution Absorbance % Fein sample per trial (mL) Dilution Volume ( 50 50 50 21 31 0,532 6...
Answer
2) To measure the concentration of phosphorous in a water sample, Phosphorous was first converted into a color complex in the presence of ammonium molybdate and SnClo. The absorbance of a series of standard phosphorous solutions was measured by a spectrophotometer and gave the calibration curve y 1.150 x, where y is the absorbance of phosphorous complex at 690 nm while x is the concentration of P in ppm. An unknown natural water sample was collected. 10.00 mL of...
Data Results (show all work on following pages) Standard Dilutions A-E Solution Absorbance А B Concentration(mg/L) 0.005 0.001 0.002 0-003 0-004 C 1. 188 0.232 0.47 0.702 0.953 D E Graph of Absorbance versus Concentration (mg/L) 81 Show work for concentration values Winai is in equation for mine samaru curve from the repuri dain? Extraction of B-carotene Mass or carrot 5.49 Absorbance of carrot solution (diluted if 0.455 necessary) Concentration of carrot solution (diluted 0.00194 if necessary)(mg/L) Concentration of original...
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