For an assay using a standard curve, why must unknown samples and standards be assayed simultaneously?
In case of standard curve, the unknown samples and standards must be simultaneously assayed as this helps in making better comparison between the two samples. It helps in calculating the absorbance and by finding out the absorbance value, it helps in finding out the value of the unknown sample. As the unknown sample and standard values are assayed together it helps in making a comparison about the values. The standard curve is basically plotting the graph from the data values of the sample and hence it is important that one is aware of the amount present in the unknown sample. As the unknown sample and standard is assayed together simultaneously, it helps in drawing the graph and also extrapolating the data
For an assay using a standard curve, why must unknown samples and standards be assayed simultaneously?
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 ...
3. Why can we assume in the Biuret assay that BSA (used in the standards) and casein (our protein of unknown concentration) give about the same amount of A550 per mg? 4. To calibrate the Ninhydrin assay, glycine was used. Draw the structures of Glycine, Arginine, and Threonine. Instead of glycine, would it be acceptable to use Arginine or Threonine? Explain!
you assay an unknown sample and its concentration falls below the linear range of your standard curve. what can you do to solve this problem?
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...
They performed a BSA assay to determine the concentration of an unknown solution X. The graph below is the BSA standard curve. In the cuvette, you add 25 ul of the unknown solution X, 200 ul of the Bradford’s reagent and 775 ul dH2O. You obtain an absorbance of 0.58 at 595nm. Use all the information to calculate the concentration of the unknown solution X. Show work Y- 0.0457x R^2- 0.9901
using my standard curve I need to determine the concentration of glucose in the unknown the equation is y=-0.1741x+0.7254 the absorbance for the unknown is 0.201
What is the minimum number of samples that could make a standard line on the calibration curve and why do we use many samples
Why must the standard curve pass through the origin (the zero point on both axes)?
In this lab, you will quantitatively estimate the protein concentration in the following 3 unknown samples: 1. Milk 2. Diet drink 3. High protein health food drink. Before you do the lab, you are expected to know the following: 1. How will you make the dilutions of the provided standard protein BSA (Bovine Serum Albumin) in the microfuge (1.7 ml small) tubes? Draw the chart for the same. 3 marks 2. How will you make the dilutions for the above...
1. During the extraction process, the protein samples must be kept on ice. Why? 2. What is the role of the bovine serum albumin (BSA) in this experiment? 3. Calculate the volume of BSA stock that will be required to make the standard solutions needed to create the BSA standard curve. Be sure to show your work and include the volume of 0.02 M phosphate buffer required to reach a final volume of 1 mL.