To prepare a gel sample, you want to load 50 ng total of protein/well. You have added200 μL of protein in 800 µL reagent and measured your sample by Bradford A595 to be 0.7 mg/mL- your dilution is unaccounted for at this point. Assuming a total final volume of 20 μL, whatvolume of protein sample, buffer and 4X SDS PAGE Loading Dye needs to be mixed to createa final sample of 50 ng protein in 20 μL?

Below answer given Explanation:Step 1: Step 2: Step 3: Step 4:Step 5:

Answered: To prepare a gel sample, you want to…

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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Using ultrafiltration, you concentrate a solution of Protein X as follows: Initial volume protein X = 300 mL Initial concentration protein X = 12 mg/mL Final volume protein X = 15 mL What's the final concentration of protein X in mg/mL?
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The SDS-PAGE protocol requires 0.5L of running buffer for the gel apparatus. The stock running buffer comes as a 20x concentrate. How should you prepare your running buffer? 50ml 20x stock + 450ml deionized water 450ml 20x stock + 50ml deionized water O 250ml 20x stock + 250ml deionized water 25ml 20x stock + 500ml deionized water 25ml 20x stock + 475ml deionized water 2ml 20x stock + 500ml deionized water O 100ml 20x stock + 400ml deionized water
A gel filtration column was calibrated by measuring elution volumes for two proteins of known mass: glutamate dehydrogenase, 290 kDa with elution volume 20.8 mL; serum albumin, 37 kDa with elution volume 42.5 mL Use these values to determine the equation of the straight line graph that relates log molar mass to elution volume. The elution volume of an unknown protein was 32.2 mL. Use the equation just determined above to estimate the molar mass of the protein in kDa units. Don't round your answer until the end to ensure your answer is within the accepted range. Round your answer to the nearest whole number (e.g. If you calculated 27.52, input your answer as 28).
what is the dna concentration suspended in PO4 buffer at ph 7.0 that has an absorbance at 260 nm equal to 1.0?
To make 100 ml 0.3% NP-40 solution for FISH post washing, how many ul 100% NP-40 solution is needed?
A protein has a molecular mass of 400 kDa when measured by size-exclusion chromatography. When subjected to gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), the protein gives three bands with molecular masses of 180, 160, and 60 kDa. When electrophoresis is carried out in the presence of SDS and dithiothreitol, three bands are again formed, this time with molecular masses of 160, 90, and 60 kDa. Determine the subunit composition of the protein and clearly explain how you came to this determination.
Figure 3: ● ● ● ● ● ● KDa ● 97.4 66.2 45.0 ● 31.0- 21.5 14.4 S-1 p-1 S-2 2-0 This figure was generated by centrifuging a pura sample of protein, removing the supernatant, and resuspending the pellet in the same volume as the supernatant to allow direct comparison. The supernatant and pellet samples were then prepared for SDS-PAGE identically and run via normal SDS-PAGE procedures. In the figure, "s" means supernatant and "p" means pellet. The text or number after the dash represents a different condition. For example, s-1 and p-1 are the supernatant and pellet samples under condition 1. It is not shown, but under wild-type conditions, essentially all of the protein is found in the supernatant. S-3 ● What does the intensity of each band represent? ● Would you find soluble protein in the supernatant or pellet? Why? Would you find aggregated protein in the supernatant or pellet? Why? For each condition (there are 5 different conditions), is there a higher percentage of the total protein…
How would you make two-fold serial dilutions such that the last tube is a 1:32 dilution of the original, concentrated material? Assume that you need to have at least 500 µl of each dilution, and you should be able to perform the dilutions in microfuge tubes with a maximum capacity of 1.5 ml.
A student performed an invertase activity assay on samples from a purification. All reactions occurred in a reaction volume of 100 µL and lasted 3.1 minutes. For each reaction, 5.00 µL enzyme was mixed with 50.0 mM sucrose in 10.0 mM acetate pH 4.8. Then 400 µL of DNS was added, the samples were heated to 90 °C for 5 minutes, and 500 µL of 50.0 mM acetate pH 4.8 was added before loading a 96 well plate in triplicate. Samples were measured at 540 nm using a path length of 0.680 cm. The fraction with the highest activity had A540 of 0.904. The standard of 14.00 mM hydrolyzed sucrose had A540 of 1.047 (the concentration given is of the 100 µL sample). The no-enzyme control had A540 of 0.174. Calculate the activity of the fraction (in nmol min-1).
A flat (not spherical) tissue engineered skin (thickness 2.5 mm) is attached to the bottom of a dish and cultured with cell media atop it (which has an oxygen concentration of 0.15 mol/m3). The diffusivity of oxygen in the skin is 3.4 E-9 m2/s, and according to experiments that you’ve performed, you want to make sure that the concentration of oxygen in the device doesn’t fall below 0.06 mol/m3. (Below that concentration harms the cells.) The rate of oxygen consumption is -5.9 E-17 mol/[cell◦s]. What is the maximum cellularity (in cells/mL, three significant digits) that this tissue engineered skin can support?
Calculate the appropriate volume (in µL) of 3X loading buffer that should be added to 24.0 µL of a sample prior to loading the sample on an agarose gel.

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