Burden, Sierra, Immunology, Tuesday C, Pre Lab 1

Burden, Sierra, Immunology, Tuesday C, Pre Lab 1 -...

Info iconThis preview shows pages 1–2. Sign up to view the full content.

View Full Document Right Arrow Icon
1 PROBLEM SET - KEY VERSION A | SPRING 2010-2011 Student Name: __Sierra Burden ____________________ _____________________ Section Day and Letter: ____Tuesday C ___________________ E-mail Address: ___scburden @stanford.edu ____________________ Part 1: Understanding the Experiment (5 points) The title should succinctly state the goal of this week’s experiment, the specific techniques performed, and the model organism used. (1 pt) Title: Using a Western Blot to identify four mouse strains based on their immune response to experimental autoimmune encephalomyelitis The objective should be written using complete sentences and be approximately ~200 words. It should provide some background, explain the purpose and techniques, discuss the variables tested (if any), and provide a hypothesis. (4 pts) Objective: The purpose of the experiment is to identify four mouse strains using a Western Blot. The Western Blot will determine the cytokines produced in an immune response to experimental autoimmune encephalomyelitis, the mouse version of multiple sclerosis. The onset of EAE depends on two factors: antigen presentation and the type T cells produced. In the experiment, two of the mouse strains B10. PL and BALB/k have MHCs that present viral peptides similar to myelin basic proteins. These strains will recognize MBP in the CNS as foreign and trigger autoimmunity. The other two strains B10.BR and BALB/c have random peptides on their MHCs. The BALB/k and BALB/c strains, however, have a mutation in the pathway that induces TH1 cytokine response. These strains produce a TH2 response as a default, unlike B10.PL and B10.PR. The positive control is a solution with cytokines and buffer, while the negative control is a solution of just buffer. The strains will be introduced to Theiler’s virus to trigger EAE. The Western Blot will then be used to identify the cytokines produced by each sample. The samples with MHC viral peptides similar to MBP and a TH1 immune response will develop EAE. The strain B10. PL, therefore, will be more likely to develop EAE. Since the strains BALB/k and BALB/c will produce TH2 cytokines instead of TH1 cytokines, EAE will not develop. Part 2: Understanding the Concepts (15 points) Lab Procedure Check-In 1. You add the gel components (gel buffer, dH2O, acrylamide mix, and the ammonium persulfate) into the flask with the correct volumes and pour the mixture into your gel pouring apparatus. Your gel never polymerizes. What happened and why did it cause that result? (1/2 pt) TEMED was not added to the gel components before pouring. TEMED would have catalyzed ammonium persulfate’s production of oxygen-free radicals. The oxygen
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Image of page 2
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 09/19/2011 for the course BIOLOGY 44Y taught by Professor Smith during the Winter '11 term at Stanford.

Page1 / 6

Burden, Sierra, Immunology, Tuesday C, Pre Lab 1 -...

This preview shows document pages 1 - 2. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online