You are running an cation exchange column at pH 7.0. Your protein has a pI of...
You are running an cation exchange column at pH 7.0. Your protein has a pI of about 8.3 and the contaminating protein has a pI of about 7.5. Which of the following statements is most true?
a. Both proteins will bind but your protein will bind more tightly
b. both proteins will bind and the contaminating protein will bind more tightly
c. neither protein will bind
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You are running an cation exchange column at pH 7.0. Your
protein has a pI of...
You have a mixture of three proteins: Protein X, MW 15 kDa, pI 4.5 Protein Y, MW 25 kDa, pI 8.5 Protein Z, MW 75 kDa,...
You have a mixture of three proteins: Protein X, MW 15 kDa, pI 4.5 Protein Y, MW 25 kDa, pI 8.5 Protein Z, MW 75 kDa, pl 5.0 2) You have a cation exchange (acidic) column, low salt (100 mM NaCl) and high salt (1 M NaCI) buffer at pH 7.0. If you loaded the above mixture of proteins on the column in low salt buffer, which proteins would most likely be retained on the column? If you loaded the...
The five proteins listed below were run on a cation exchange column at pH = 7...
The five proteins listed below were run on a cation exchange column at pH = 7 Cytochrome C pl = 10.7 Ribonuclease A pl = 9.5 Myoglobin pl = 7.4 Carbonic anhydrase pl = 6.0 Glucose oxidase pl = 4.2 Which of the following statements is TRUE? A) Glucose oxidase and carbonic anhydrase will be separated in the low salt wash b) Cytochrome C binds and then elutes first when the salt concentration is increased c) decreasing the pH will...
Let's say that an unknown protein has an isoelectric point of 7.0. a) Will this protein...
Let's say that an unknown protein has an isoelectric point of 7.0. a) Will this protein be positively or negatively charged at pH 4.0? b) Will this protein be an anion or a cation at pH 10.0? c) If you had at your disposal a cation exchange column, would you separate the protein from others in the mix on the basis of cation exchange or anion exchange? Explain. d) Would phosphate buffer (pKa 7.20), ethanolamine buffer (pKa 9.50), or citric...
You have a mixture of the proteins listed below. Protein pI Molecular Weight (kDa) A 3.1...
You have a mixture of the proteins listed below. Protein pI Molecular Weight (kDa) A 3.1 265 B 6.9 93 C 10.3 96 D 7.1 43 E 8.6 189 1. You load the protein mix onto a cation exchange column at pH 5. Next, you apply a "washing" step by passing through buffer at pH 5. Finally, for your elution step, you apply a pH gradient starting from pH 2.0 to pH 13.0. (A gradient buffer system allows you to...
NOTES: To separate on a size-exclusion column, differences in MW need to be greater than 10%....
NOTES: To separate on a size-exclusion column, differences in MW need to be greater than 10%. To separate proteins by charge, the difference in pI must be at least 0.5 pH unit. Consider the table of four proteins below: Protein Size pI Size exclusion Charge at pH 7.4 DEAE-elution 1 49 kDa 6.8 2 51 kDa 8.1 3 53 kDa 6.4 4 99 kDa 8.3 5 106 kDa 3.8 6 143 kDa 7.9 (a) In what order would the proteins...
Peptides can be separated using an ion-exchange column based on their isoelectric (pl) values. At which...
Peptides can be separated using an ion-exchange column based on their isoelectric (pl) values. At which pH values would two different peptides, one with a pl of 5.0 and the other with a pl of 9.2, bind to a cation- and anion-exchange column? Each peptide may be capable of binding to each column at more than one pH value. anion-exchange column at pH = 3.0 cation-exchange column at pH = 3.0 Answer Bank peptide B pI = 9.2 peptide A...
(3 points) A Student uses cation exchange chromatography to separate Protein 1 pi9.5) and Protein 2...
(3 points) A Student uses cation exchange chromatography to separate Protein 1 pi9.5) and Protein 2 pl 45). The column is equilibrated using a 20m phosphate buffer, pH 6.0 and then the sample containing Protein 1 and Protein 2 is applied to the column. After the sample is applied to the column, the column is first washed with a low salt buffer. Subsequently, the column is washed with a high salt buffer. The elution profile is shown below. Clearly indicate...
Please EXPLAIN your answer If protein A has a pl of 3.1, protein B has a...
Please EXPLAIN your answer
If protein A has a pl of 3.1, protein B has a pl of 6.8, and protein C has a pl of 8.9, which protein would elute FIRST from a cation exchange column at pH 7? н H R-C-COOH Z R R -C-C00“ -C-000 +NH + NH3 NH2 Net charge: +1 Cationic form Zwitterion (pl) Anionic form protein C pl = 8.9 pH <pl cationic protein B pl = 6.8 pH = pl neutral protein A...
______ 2. Using the accompanying table, determine which protein would stick to a carboxymethyl cellulose ion...
______ 2. Using the accompanying table, determine which protein would stick to a carboxymethyl cellulose ion exchange column (the column has stationary carboxyl groups) at pH 7.0. a) glucose oxidase b) horse myoglobin c) ribonuclease A d) cytochrome C Protein Isoelectric Point Amyloglucosidase Glucose oxidase Trypsin inhibitor β-Lactoglobulin Carboanhydrase Myoglobin horse Lentil lectin Ribonuclease A Cytochrome C pI 3.5 pI 4.2 pI 4.5 pI 5.15/5.3 pI 6.0 pI 7.35 pI 7.75/8.0/8.3 pI 9.45 pI 10.65
At a pH of 7.0 predict what will happen to vasopressin (in its uncapped/free terminal amine,...
At a pH of 7.0 predict what will happen to vasopressin (in its uncapped/free terminal amine, but oxidized form) if... a) you applied a cell lysate containing vasopressin to an anion exchange column (i.e. would it stick)? How might you exploit this column (e.g. by adjusting pH, e.g. pH 4, pH 11, and running specimen over the column again) to increase its purity? b) Predict what would happen to vasopressin if you attempted to purify it from your lysate using...
You have a mixture of three proteins: Protein X, MW 15 kDa, pI 4.5 Protein Y, MW 25 kDa, pI 8.5 Protein Z, MW 75 kDa, pl 5.0 2) You have a cation exchange (acidic) column, low salt (100 mM NaCl) and high salt (1 M NaCI) buffer at pH 7.0. If you loaded the above mixture of proteins on the column in low salt buffer, which proteins would most likely be retained on the column? If you loaded the...
Peptides can be separated using an ion-exchange column based on their isoelectric (pl) values. At which pH values would two different peptides, one with a pl of 5.0 and the other with a pl of 9.2, bind to a cation- and anion-exchange column? Each peptide may be capable of binding to each column at more than one pH value. anion-exchange column at pH = 3.0 cation-exchange column at pH = 3.0 Answer Bank peptide B pI = 9.2 peptide A...
(3 points) A Student uses cation exchange chromatography to separate Protein 1 pi9.5) and Protein 2 pl 45). The column is equilibrated using a 20m phosphate buffer, pH 6.0 and then the sample containing Protein 1 and Protein 2 is applied to the column. After the sample is applied to the column, the column is first washed with a low salt buffer. Subsequently, the column is washed with a high salt buffer. The elution profile is shown below. Clearly indicate...
Please EXPLAIN your answer
If protein A has a pl of 3.1, protein B has a pl of 6.8, and protein C has a pl of 8.9, which protein would elute FIRST from a cation exchange column at pH 7? н H R-C-COOH Z R R -C-C00“ -C-000 +NH + NH3 NH2 Net charge: +1 Cationic form Zwitterion (pl) Anionic form protein C pl = 8.9 pH <pl cationic protein B pl = 6.8 pH = pl neutral protein A...
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