A spherical cell with the diameter of 10uMhas a protein concentration of 20 mg/ml. Determine the number of protein molecules within the cell if the molecular weight of an average protein is 50,000 daltons (g/mol). Recall that Avogadro's number is NA 6.0221367×1023 molecules/mol. =
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- Suppose the major solutes in intact lysosomes are KCl (~0.1 M) and NaCl (~0.03 M). When isolating lysosomes, what concentration of sucrose is required in the extracting solution at room temperature (25oC) to prevent swelling and lysis? Calculate how much sucrose (342.3 g/mol) would you need to make 500 ml of this extracting solution.How many copies of a protein need to be present in a cell in order for it to be visible as a band on an SDS gel? Assume that you can load 100 µg of cell extract onto a gel and that you can detect 10 ng in a single band by sil ver staining the gel. The concentration of protein in cells is about 200 mg/mL, and a typical mammalian cell has a volume of about 1000 μm³ and a typical bacterium a vol ume of about 1 µm³. Given these parameters, calculate the number of copies of a 120-kd protein that would need to be present in a mammalian cell and in a bacterium in order to give a detectable band on a gel. You might try an order-of-magnitude guess before you make the calcula tions.Student A says that he can insert any value of optical density in the equation of standard curve to determine the corresponding value of protein content. Does the student say it right or wrong? Explain why?
- Please answer the question below and show all your work. You are given a pure protein sample to characterize and provided the following information: Its molar extinction coefficient, ε280, is 0.25 liters micromole^-1 cm^-1 Using a 0.5 cm pathlength cell, you measure the absorbance at 280 nm of a 20- fold dilution of your pure protein in solution (by this, we mean that 50 ul of the protein sample was diluted to a final volume of 1 ml) and find A280 = 0.40. What is the original concentration of the protein before dilution?Give only typing answer with explanation and conclusion You want to make 94 µL of the diluted Cell-Free extract for estimation of total protein. To do this, how much water will you add to the correct volume of the undiluted cell-free extract? Note: Cell-free extracts will need to be diluted 1:25 in water.Calculate θ for a certain protein-ligand pair when the ligand concentration = 1 M and the Kd = 1 X 10-15 M.
- You are given a pure protein sample to characterize and provided the following information: Its molar extinction coefficient, ε280, is 0.25 liters micromole-1 cm-1 in both the folded and unfolded form Its ΔGo for unfolding is 1.5 kcal/mol at 37o (where RT = 0.59 kcal/mole) A) Using a 0.5 cm pathlength cell, you measure the absorbance at 280 nm of a 20-fold dilution of your pure protein in solution (by this, we mean that 50 ul of the protein sample was diluted to a final volume of 1 ml) and find A280 = 0.40. What is the original concentration of the protein before dilution? B) What is the concentration of the unfolded form of the protein in your sample?Predict the number of bands and apparent mol. wt. of the following proteins on SDS gels. 1. A trimeric protein containing three chains, each with a molecular weight of 60,000 Da (60 kDa).The graph below is a standard curve generated by plotting the distance travelled by the size standards on an SDS-PAGE gel. If a protein band moved to a distance of 2 cm, the approximate Mol. Wt. of that protein is 17 KDa 70 KDa Not enough information 100 KDa
- Protein concentration can readily be determined using the Beer-Lambert law: A = e l c where A = absorbance e = molar absorption coefficient (M-1cm-1) l = light path length (cm) c = concentration (M) If the molar absorption coefficient at 280 nm for yeast ADH is 48860 M-1cm-1 and a 10 mL solution of the protein has an absorbance at 280 nm of 0.4 (as measured by a spectrometer with pathlength 1 cm), then what is the concentration of the protein solution (in μM)? i.e. concentration = ______ μM If the molecular weight of the protein is 36849, what is its concentration in mg/mL? i.e. concentration = _______ mg/mL For each part of the question, show your calculations to arrive at your answers.To get an idea of the different sizes of various cellular components, do the following calculations: Assume that the cell, its nucleus, and a globular protein – for example, and enzyme – are spherical. In addition, assume the diameter of the protein is 5 nm (nanometers), the diameter of the cell is 100 μm (micrometers), and the diameter of the nucleus is 40 μm. You may have to review your metric conversions! If you draw the globular protein as a sphere with a diameter of 2 cm (approximately the diameter of a U.S. penny), what size would each of the following measurements of the cell be if drawn to the same scale (5 nm real length = 2 cm)? a. The radius of a microtubule, 15 nm b. The diameter of the nucleus c. The diameter of the cell d. The volume (V = 4/3 ∏r3) of the globular protein, in decimal form to the hundredths place. e. The volume of the nucleus f. The volume of the cell0.0 M sucrose = _______ 0.008_____________ g/min 0.4 M sucrose = _______ -0.013_____________ g/min 0.6 M sucrose = ________ -0.023g____________ g/min 0.8 M sucrose = ________-0.026____________ g/min 1.0 M sucrose = ________ -0.033____________ g/min Can i please have the step by step calculations Using the osmosis data in the question above, determine the percent weight change for each sucrose solution . Write your answer in standard notation and use 1 digit past the decimal point - e.g. 8.6. (Note that the last zero in the answer is considered to be a digit) 0.0 M sucrose = _________ ___________% 0.4 M sucrose = _______ ______________% 0.6 M sucrose = _____________________% 0.8 M sucrose = __________________% 1.0 M sucrose = ____________________%