You have obtained 150 µL of a protein sample of which you want to perform a...
You have obtained 150 µL of a protein sample of which you want to perform a Bradford Assay. You perform a 10-fold dilution of your sample and use 10 µl of this dilution to quantify the amount of protein using a Bradford assay. You obtain an absorbance value of 0.63. Using the standard curve and equation you obtained in class, determine the concentration of protein (µg/µL) of your original sample. Show your work.
equation: y=0.0139x-0.0153
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You have obtained 150 µL of a protein sample of which you want
to perform a...
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...
1. You want to prepare a 1:50 dilution of your protein extract in a total volume...
1. You want to prepare a 1:50 dilution of your protein extract in a total volume of 1000 uL. You will need ___ uL of protein extract and ___ uL of water. 2. You are provided with a solution of BSA that is 100 ug/mL and make a 10-2 dilution? What is the resulting concentration of the DILUTED BSA? 3. Which of the following statements about enzymes is true? Select one: a. Enzymes increase the rate of a chemical reaction...
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 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
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 ...
You have purified a sample of your protein of interest and have measured the A280 of a 1 in 3 dilution of the protein sample (1 cm pathlength). The value you have obtained for A280 was 0.576. Given th...
You have purified a sample of your protein of interest and have measured the A280 of a 1 in 3 dilution of the protein sample (1 cm pathlength). The value you have obtained for A280 was 0.576. Given that you do not know the sequence of your protein, calculate the approximate concentration of your original protein sample in mg/mL. Give your answer to 2 s.f.
You only have 150 pl stock solution that contains 100 mgml of protein. You want to...
You only have 150 pl stock solution that contains 100 mgml of protein. You want to make a set of standards with the concentrations shown below. You need a minimum of 500 l of each standard left after you have finished all your dilutions. Using the technique of serial dilution, explain how you would make your standards by filling in the chart below include the final dilution of the standard as compared to the original stock concentration and the final...
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...
One of the common methods to quantify DNA is to measure A260 (UV absorbance at 260...
One of the common methods to quantify DNA is to measure A260 (UV absorbance at 260 nm) of DNA solution using a spectrophotometer. You took 10 µl of your plasmid DNA preparation and dilute with 990 µl of H2O. The A260of this diluted solution was 0.30. Calculate the concentration of your plasmid DNA preparation (before the dilution). It has been established that 50 µg/ml of double-stranded DNA gives A260 = 1.0. My plasmid DNA preparation is _____________________ mg/ml. From this...
how do you properly come up with the answers to the highlighted questions? rebnuary 25,2019IO 9)...
how do you properly come up with the answers to the
highlighted questions?
rebnuary 25,2019IO 9) (10 points) You are trying to determine if the amount of protein the manufacturer says it supplies in baby formula is actually correct. Using the Bradford assay, you have constructed the standard curve shown below. You make a 1:100 dilution of baby formula in phosphate buffer. You then measure the absorbance of 10, 20, or 40 μ1 of the dilution solution. ao)xcio) xo) 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...
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 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
You only have 150 pl stock solution that contains 100 mgml of protein. You want to make a set of standards with the concentrations shown below. You need a minimum of 500 l of each standard left after you have finished all your dilutions. Using the technique of serial dilution, explain how you would make your standards by filling in the chart below include the final dilution of the standard as compared to the original stock concentration and the final...
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...
how do you properly come up with the answers to the
highlighted questions?
rebnuary 25,2019IO 9) (10 points) You are trying to determine if the amount of protein the manufacturer says it supplies in baby formula is actually correct. Using the Bradford assay, you have constructed the standard curve shown below. You make a 1:100 dilution of baby formula in phosphate buffer. You then measure the absorbance of 10, 20, or 40 μ1 of the dilution solution. ao)xcio) xo) Sample...
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