E11 Lab Report-Aastha Ramolia

docx

School

University of Texas, Dallas *

*We aren’t endorsed by this school

Course

2281

Subject

Biology

Date

Dec 6, 2023

Type

docx

Pages

Uploaded by aastharamolia2003

Experiment 11: Protein Agarose Gel Electrophoresis Total Points: 20pts Your Name:_Aastha Ramolia_ Section: _304_NetID:__axr200124__ Note: You do not need a title sheet for this report. Type your answers and upload the completed report file (with questions) in eLearning by the end of your lab session. E11: Protein Agarose Gel 1) Present the photos (or illustration) of two protein gels after electrophoresis (Part A and B). Label each lane by typing the name of the sample loaded. Label the cathode and the anode along with the positions of the wells on the correct end. Illustration of Gel electrophoresis with pH 8.6 buffer (2 points) Cathode Anode Wells 4.Serum Albumin 3.Hemoglobin 2.Myoglobin 1.Cytochrome 1

Illustration of Gel electrophoresis with pH 9.2 buffer (2 points) 2) The pH of the Tris-Glycine buffer used in Part A is 8.6, what is the net charge on the cathode Anode wells 2

protein molecules in the samples? Which electrode should each of the four protein samples migrate toward during the electrophoretic separation? (4 points) The net charge for Serum albumin is negative (-3.8) and it migrates toward the anode. Cytochrome C has a net charge is positive (1.6) and it migrates toward the cathode. Hemoglobin has a net charge is negative (-1.8) and it migrates toward the anode. Myoglobin has a net charge is negative (-1.4) and it migrates toward the anode. 3) Explain why did Serum Albumin migrate faster than myoglobin and Hemoglobin in Part A? (1 point) Serum Albumin migrated faster because its pI of 4.8 is lower than Hemoglobin’s pI of 6.8 and Myoglobin’s pI of 7.2. The net charge of Serum Albumin is more negative than the net charges of Hemoglobin and myoglobin in pH 8.6 buffer, which makes Serum albumin more negatively charged compared to the other samples. 3

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) How would the separation of the four proteins in Part A change if the students mistakenly used the pH 9.2 buffer instead of the pH 8.6 buffer? Would the samples migrate to different electrodes? How would the migration rates change? (3 points) The samples would The samples would not migrate towards different electrodes; however, their migration rates would change. The Cytochrome would migrate slower, it will have a smaller positive net charge in a 9.2 pH buffer than in an 8.6 pH buffer. The Myoglobin, Hemoglobin, and Serum Albumin samples would all migrate faster because they would have larger negative net charges in the 9.2 pH buffer than the 8.6 pH buffer. 5) In Part B, the pH of the Tris-Glycine buffer used is 9.2 and the pI of normal human hemoglobin (HbAHbA) is 6.9. W hat is the net charge on normal human hemoglobin (HbAHbA), heterozygous (HbAHbS) and sickle cell hemoglobin (HbSHbS)? Which electrode should each sample migrate toward during the electrophoretic separation? (2 points)) The net charge for normal human hemoglobin (HbAHbA), is negative (-2.3) and it would migrate towards the anode. The net charge for heterozygous (HbAHbS) is negative and it will migrate towards the anode. The net charge for sickle cell hemoglobin (HbSHbS) is negative, therefore it will move towards the anode. 4

6) Which amino acid residue replaces glutamic acid in sickle cell hemoglobin (HbSHbS)? What is the pI and the migration rate of HbSHbS relative to HbAHbA at pH 9.2 in Part B? (2 points) A single glutamic acid is replaced by valine. The pI of valine is higher than glutamic acid, making the overall pI of sickle cell hemoglobin higher than that of normal human hemoglobin (6.9). Relative to normal human hemoglobin, the sickle cell hemoglobin will have a slower migration rate in 9.2 pH buffer, because the net charge is less negative than the net charge of normal human hemoglobin. 7) How would the separation of the three hemoglobin proteins change If Part B of the experiment was carried out at pH 8.6 instead of pH 9.2? Would the samples migrate to different electrodes? How would the migration rates change? (2 points) The three hemoglobin proteins will still migrate towards the anode, but their migration rates would be slower. Because the net charge for each of the samples would be less negative therefore the samples would migrate at a slower rate in 8.6 pH buffer. 5

8) Use your biology textbook as a guide, hand-draw one sample structure of the following types of amino acids: (2points, structures copied and pasted are accepted in Fall 2023 ) acidic amino acid basic amino acid non-polar amino acid polar, uncharged amino acid 6

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

MEGA Tetrapods-Danielle Clements - Tagged.pdf

Danielle Clements- Ecological Modeling.pdf

Dichotomous Key Cells & Viruses- Post Lab questions and photos.docx

EVR1001L_Fall_2023_Lab_4_(1).docx

Lab 13. DNA Biology and Technology.docx

BIOL 1108L Field Survey Make Up Assignment 2.docx

Analysis & Visualization Project (written).docx

Lab5Assignment_BIO120F2023.docx

Lab 2 Manual.docx

Renal Case Study.docx

Lung Volumes and Capacities.docx

Ans_ProbSet8_Ch9,10.docx

Recommended textbooks for you

Biochemistry

ISBN:9781305577206

Author:Reginald H. Garrett, Charles M. Grisham

Publisher:Cengage Learning

Comprehensive Medical Terminology

Nursing

ISBN:9781133478850

Author:Jones

Publisher:Cengage

Principles Of Pharmacology Med Assist

Biology

ISBN:9781337512442

Author:RICE

Publisher:Cengage

Basic Clinical Laboratory Techniques 6E

Biology

ISBN:9781133893943

Author:ESTRIDGE

Publisher:Cengage

Biomedical Instrumentation Systems

Chemistry

ISBN:9781133478294

Author:Chatterjee

Publisher:Cengage

SEE MORE TEXTBOOKS

Recommended textbooks for you

Biochemistry
Biochemistry
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Cengage Learning
Comprehensive Medical Terminology
Nursing
ISBN:9781133478850
Author:Jones
Publisher:Cengage
Principles Of Pharmacology Med Assist
Biology
ISBN:9781337512442
Author:RICE
Publisher:Cengage
Basic Clinical Laboratory Techniques 6E
Biology
ISBN:9781133893943
Author:ESTRIDGE
Publisher:Cengage
Biomedical Instrumentation Systems
Chemistry
ISBN:9781133478294
Author:Chatterjee
Publisher:Cengage