Show the order of elution from a cation-exchange resin using a buffer pH of 4.5: aspartic acid, valine, arginine, and alanine. Provide a brief explanation.
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Show the order of elution from a cation-exchange resin using a buffer pH of 4.5: aspartic acid, valine, arginine, and alanine. Provide a brief explanation.
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- 4. A solution containing egg albumin (pl-4.6), B-lactoglobulin (pl-5.2), and chymotrypsinogen (pl=9.5) was loaded onto a column of diethylaminoethyl cellulose (DEAE-cellulose) at pH 5.4.The column was then eluted with a pH 5.4 buffer, with an increasing salt concentration. Predict the elution pattern.Choose among proteins A, B, C, and D. 1. Protein that will most strongly bind to an anion exchange colum. 2. Protein that will elute last in gel filtration chromatography. 3. Protein that will elute first in hydrophobic interaction chromatography. 4. Protein that will elute last in a carbohydrate containing colum.1.2. Using DEAE-cellulose as ion exchange resin, indicate the starting and ending pH for the narrowest experimental pH range used to separate an amino acid mixture consisting of Cys, His and Leu Starting pH: _____ Ending pH: _____
- Explain the Rf value obtained on the basis of the structure of amino acids use. Explain in 1-3 sentence only.Consider a crude extract with a mixture of the 5 proteins listed below. Protein A 4.5 pl 160 molecular weight, kDa Protein B 12.5 pl and 65 molecular weight Protein C 5.0 pl, 15 molecular weight Protein D 6.8 pl, 150 molecular weight Protein E 9.5 pl, 45 molecular weight You load this protein mixture onto an anion exchange column at pH 11. Next, you apply a "washing" step by passing through buffer at pH 11. Finally, for your elution step, you apply a pH gradient starting from pH 11 to pH 2.0 (A gradient buffer system allows you to gradually and continuously change the pH of your mobile phase starting from pH 11 up to pH 2). You load this same protein mixture onto a Size Exclusion column. Please indicate the order in which these proteins will elute for both. Group of answer choices Yes or No, please explain your answer. If your protein of interest is protein A, would using anion exchange column be completely successful at separating it from all the other proteins? Group of answer…In a mixture of five proteins listed, draw an elution profile (Absorbance vs. mL eluted, arbitrary) for the purification of the listed proteins on a gel filtration chromatography resin: cytochrome c (pI = 5.4; Mr = 13,000), immunoglobulin G (pI = 7.3; Mr = 145,000), ribonuclease A (pI = 9.6; Mr = 13,700), RNA polymerase (pI = 6.3; Mr = 450,000), human serum albumin (pI = 5.4; Mr = 68,500). Label your elution peaks. Draw a sketch of an SDS-PAGE, reflecting the mobility of the above mixture as they elute from the column. Label you protein bands.
- Define a chaotropic agent. What effect would a chaotropic agent have on protein structure?You are given 0.5 M solution of the amino acid Arginine. pK1 (α-carboxyl group) = 2.17 pK2 (α- amino group) = 9.03 and pK3 (guanidine group) = 12.48 Determine the pH of the solution if you add 15 mL 0.25 M HCl to 10 mL of the 0.5 M Arginine. Assume that Arginine is in isotonic state. Show all calculations b. Draw the structure of the amino acid at the pH determined in question 1You are given 0.5 M solution of the amino acid Arginine. pK1 (α-carboxyl group) = 2.17 pK2 (α- amino group) = 9.03 and pK3 (guanidine group) = 12.48 Determine the pH of the solution if you add 15 mL 0.25 M HCl to 10 mL of the 0.5 M Arginine. Assume that Arginine is in isotonic state. Show all calculations
- Calculate the concentration of 0.2ml amino acid when path length is 1cm and has an absorbance value of 0.50, if a 0.5ml solution of same amino acid has a concentration of 0.72mg/ml with an absorbance of 0.9.In a 0.1000 M acetic acid solution at 25 degrees celsius , the acid ionizes to the extent of about 1.34 %. Since each molecule of acetic acid which ionizes produces 1 H+ ion and 1 C2H3O2- ion, the concentration in the solution are: HC2H3O2 < -----------> H+ + C2H3O2-Given the amino acids below, what amino acid will elute first in a cation-exchange column. 1. Aspartate or Lysine using a buffer at pH 7 2. Arginine or Methionine using a buffer at pH 7 3. Lysine or Valine using a buffer at pH 7 4. Glycine or Valine using a buffer at pH 7 5. Serine or Alanine using a buffer at pH 7