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- r^2=6Dt , where D is the diffusion coefficient of thediffusing object and t is the time that the object is allowed to diffuse. If the diffusion coefficient, D, of a small protein is 5 x 10^-10 m2 s-1, how long (on average) does ittake for the protein to diffuse across a parasitic wasp that is 189 um long ?After blood collection, the red cells are separated from the serum to be used for the preparation of the stock solution. How is it done? For the serial dilution, your stock solution must have a concentration of 3.5 mg/mL. How much diluent must be added to the 5.3 mg/mL red cell to prepare the stock solution? Show pertinent solution/s. What is the appropriate diluent used for the preparation of the red cell suspension?In step 3 of today's protocol, it mentions that the incubation step with the P2 buffer should not be allowed to proceed for more than 5 minutes. a) What is the chemical composition and concentration (not mass) of each component of the P2 buffer?b) c) What is the purpose of adding the P2 buffer? What would be the major problem if this step is allowed to proceed for a long period of time?
- Compare the lysis for bacteria to that for CHO (Chinese Hamster Ovary) cells. Note, CHO cells are commonly used for production of therapeutics. How resistant are CHO cells compared to the bacterial cells? If the same level of disruption is expected for CHO cells and the homogenizer is operated at the same pressure for both types of cells, would you expect CHO cells to require more or less number of passes compared to E.coli or Bacillis brevis cells?In 1925, E. Gorter and F. Grendel used an apparatus like that described in Problem 1 to determine the surface area of a lipid monolayer formed bylipids extracted from erythrocytes of several animal species. They used a microscope to measure the dimensions of individual cells, from which they calculated the average surface area of one erythrocyte. They obtained the data shown in the table below. Were these investigators justified in concluding that “chromocytes [erythrocytes] are covered by a layer of fatty substances that is two molecules thick” (i.e., a lipid bilayer)?2. What are the substances added in the culture media utilized by the organism producing hydrogen sulfide? Give 2 other media used for the production of hydrogen sulfide. 3. Explain the mechanism taking place in the hydrolysis of urea which leads to the formation of bluish red color. 4. Discuss why human blood plasma will not always yield reliable results.
- Here is a chloride cell in the gill epithelium of a fish. For reference, NKA = Na+/K+ ATPaseNKCC = Na+/K+/Cl- cotransporter. Given that the chloride cells of salt water fish have the NKCC transporter on the basal (bottom) side of the epithelium, what do you predict regarding the NKCC transporter in chloride cells of freshwater fish? A. The transporter must move chloride in the opposite direction, also from the bottom side of the cell. B. The NKCC transporter, as localized in salt water fish, could also work in freshwater fish. C. The transporter must move chloride in the opposite direction, but from the apical (top) side of the cell. D. A transporter is not needed to move Cl- ions into cells from fresh water.The following observations are made on an unknownmembrane protein, X. It can be extracted from disrupted erythrocyte membranes into aconcentrated salt solution, and it can be cleaved into fragments by proteolytic enzymes. Treatment of erythrocytes with proteolytic enzymes followed by disruption and extraction of membrane components yields intact X. However, treatment of erythrocyte “ghosts” (which consist of just plasma membranes, produced by disrupting the cells and washing outthe hemoglobin) with proteolytic enzymes, followed by disruption and extraction, yields extensively fragmented X. What do these observations indicate about the location of X in the plasma membrane? Do the properties of X resemble those of an integral or peripheral membrane protein?1. What 2 controls do your laboratory instructions ask you to use when using the refractometer? 2. On a Urinalysis strip what is the difference between a speckled pattern and a homogenous pattern for a positive reaction on the blood reaction pad? 3. If the following analytes are positive on the reagent strip, what should we expect to see microscopically? Blood (speckled pattern):________________________________ Blood (homogenous pattern):________________________________ Leukocyte esterase:__________________________ Glucose:________________________ Bilirubin with positive icotest:______________________________
- Here is a chloride cell in the gill epithelium of a fish. For reference, NKA = Na+/K+ ATPaseNKCC = Na+/K+/Cl- cotransporter. (image 1) The same proteins have been identified in shark rectal gland, marine birds and reptiles (salt glands in nostrils), marine fishes (chloride cells in their gills) and mammals that transport salt in their kidneys. (image 2) When biologists were testing the mechanism of salt excretion in sharks, they used a chemical called ouabain to inhibit the Na+/K+ ATPase to see if there was an effect. Which result would you expect to see with ouabain treatment? A. A decrease in Cl- in the epithelial cells. B. An increase in ADP in the epithelial cells. C. An increase in K+ in the epithelial cells. D. A decrease in Na+ in the epithelial cells.From this standard curve and chart below, does the separation of molecules in the mixture appear successful from the gel filtration? Is there a clearlydefined separation between molecules? Explain your conclusions. Parameters required for calculation of coefficient (Kd) for unknown protein Volume eluted (mL) Which variable does this volume represent in the equation for Kd? Fraction with maximal DNP-Aspartate detected 36 Vt Fraction with maximal Protein detected 24 Ve Fraction with maximal Blue dextran detected 6 VoPropose a serial dilution that would give total dilution of 1/158000000 (six zeros) Each tube in your dilution series must have a volume of 50 ml or less.