Recitation worksheet #3

docx

School

Baylor University *

*We aren’t endorsed by this school

Course

4341

Subject

Biology

Date

Feb 20, 2024

Type

docx

Pages

6

Uploaded by BarristerFreedomApe35

Report
Agenda: Leh. Ch. 3 (VVP Ch. 5) Working with proteins Stretch (1) 5 min. Heavy weight lifting (4) 20 min. Cool Down (1) 5 min. RS TA Worksheet #3 Monday Sep 13 th to 17 th Stretch: 1) Work in a small group and complete the table (can be done at home) Technique Protein properties Conditions for binding and elusion Ion exchange chromatography SDS-PAGE Isoelectric focusing Size-exclusion chromatography Affinity chromatography
Heavy weightlifting 2) The process for protein A purification was tracked by SDS-PAGE and activity measurements. (Complete during recitation, about 10 min) A) Using the information provided, find the specific activity and purification factor for each step. B) Which purification step was the least successful? C) Which purification step was the most successful? D) Why is it important to run an SDS-PAGE in conjunction with specific activity analysis? Procedure Total protein (mg) Activity (units) Specific activity Purification factor Homogenate 2368 3482 Salting out 1017 3356 Ion exchange chromatography 272 3046 Size-exclusion chromatography 56 2890 Affinity chromatography 6.0 2490
3) You are working as an undergraduate laboratory assistant and the postdoc in charge of the lab hands you a table with details about a protein mixture. He instructs you to return three separate vials, each containing pure fractions of the proteins. Additionally, he instructs you to only use the solutions and equipment listed below. (Hint: imidazole can bind to nickel (Ni 2+ ) very tightly) (Complete during recitation, 15min) Table: Oligopeptide pI Average M.W. NYLVDCAITGWDESALIA HHHHHHGILVNQYWPCF G RGIKPCRGIKPWRGIKPD Laboratory materials: Cation exchange chromatography column Size-exclusion chromatography column Ni 2+ affinity chromatography column Combo one: Buffer pH=6 [NaCl]= 25mM Buffer pH=6 [NaCl]= 125mM Combo two: Buffer pH=8.5 [NaCl]= 25mM Buffer pH=8.5 [NaCl]= 125mM Combo three: Buffer pH= 8.5 [NaCl]= 125mM Buffer pH= 8.5 [NaCl)= 125mM [imidazole]= 500mM List in chronological order the steps you will take to achieve separation and list which solutions and equipment you use in each.
Your preview ends here
Eager to read complete document? Join bartleby learn and gain access to the full version
  • Access to all documents
  • Unlimited textbook solutions
  • 24/7 expert homework help
4) Using the experimental results shown below answer the following questions (complete during recitation, 10min) a) Name the two dimensions used to separate proteins in this experiment. b) Which proteins have the exact same pI? c) Which protein have the same molecular weight? d) If you take the original protein sample used to run this gel and load it in a size- exclusion column, what would be the order of elution? 5) You wish to determine the sequence of a short peptide. (Can be completed at home using video lecture or reference to page 92, proteases) Cleavage with chymotrypsin yields three peptides with the sequences: AGKQLEDGRTLSDY, IPPDQQRLIF NIQKESTLHLVLRLRGGMQIF Cleavage of the peptide using a different enzyme, trypsin yields seven oligopeptides with the sequences: LIFAGK GGMQIF QLEDGR LR ESTLHLVLR TLSDYNIQK IPPDQQR Find the sequence of the original peptide
Cool down 6)The mass of each amino acid residue is shown below. (Can be completed at home using video lecture as a reference) Residu e Mass (D) Residue Mass (D) A 71.0 L 113.1 R 156.1 K 128.1 N 114.0 M 131.0 D 115.0 F 147.1 C 103.0 P 97.1 Q 128.1 S 87.0 E 129 T 101.0 G 57.0 W 186.1 H 137.1 Y 163.1 I 113 V 99.1 A) Explain why mass spectrometry cannot distinguish Leu and Ile? B) In mass spectrometry sequencing, not all peptide bonds are broken, if cleavage between two Gly resides does not occur, which amino acid would be identified in place of the two glycine amino acids? C) What amino acid would be identified if a bond between Ser and Val did not break? 7) What protein information can you gain from tandem mass spectrometry?
Your preview ends here
Eager to read complete document? Join bartleby learn and gain access to the full version
  • Access to all documents
  • Unlimited textbook solutions
  • 24/7 expert homework help

Related Documents

Chapter_012-1.docx
Chapter_011-1.docx
2023 Week 4 Sordaria Prelab.pdf
Chapter_013.docx
pierce_genetics7e_solutionsmanual_ch04.docx
pierce_genetics7e_solutionsmanual_ch05.docx
Micro exam 5.pdf
Physicology.pdf
Lab Quiz 2.pdf
Lab 3 Final, T6 update.pdf
Chap12 Exam Review.docx
Ch 2 Self-Paced Study Medterm.docx
Recommended textbooks for you
Text book image
Principles Of Pharmacology Med Assist
Biology
ISBN:9781337512442
Author:RICE
Publisher:Cengage
Text book image
Anatomy & Physiology
Biology
ISBN:9781938168130
Author:Kelly A. Young, James A. Wise, Peter DeSaix, Dean H. Kruse, Brandon Poe, Eddie Johnson, Jody E. Johnson, Oksana Korol, J. Gordon Betts, Mark Womble
Publisher:OpenStax College
Text book image
Biology 2e
Biology
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:OpenStax
Text book image
Biology: The Unity and Diversity of Life (MindTap...
Biology
ISBN:9781337408332
Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Publisher:Cengage Learning
Text book image
Biology (MindTap Course List)
Biology
ISBN:9781337392938
Author:Eldra Solomon, Charles Martin, Diana W. Martin, Linda R. Berg
Publisher:Cengage Learning
Text book image
Curren'S Math For Meds: Dosages & Sol
Nursing
ISBN:9781305143531
Author:CURREN
Publisher:Cengage
SEE MORE TEXTBOOKS
Recommended textbooks for you
Text book image
Principles Of Pharmacology Med Assist
Biology
ISBN:9781337512442
Author:RICE
Publisher:Cengage
Text book image
Anatomy & Physiology
Biology
ISBN:9781938168130
Author:Kelly A. Young, James A. Wise, Peter DeSaix, Dean H. Kruse, Brandon Poe, Eddie Johnson, Jody E. Johnson, Oksana Korol, J. Gordon Betts, Mark Womble
Publisher:OpenStax College
Text book image
Biology 2e
Biology
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:OpenStax
Text book image
Biology: The Unity and Diversity of Life (MindTap...
Biology
ISBN:9781337408332
Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr
Publisher:Cengage Learning
Text book image
Biology (MindTap Course List)
Biology
ISBN:9781337392938
Author:Eldra Solomon, Charles Martin, Diana W. Martin, Linda R. Berg
Publisher:Cengage Learning
Text book image
Curren'S Math For Meds: Dosages & Sol
Nursing
ISBN:9781305143531
Author:CURREN
Publisher:Cengage
SEE MORE TEXTBOOKS