K-12 Programs

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Immunology Field Trip
Set-up

Note: Set up 6 students per bench.

I. IgY Purification

Reagents needed:

1. One egg per bench.
2. Precipitation buffer A (ppt A): 8 ml per tube; 2 tubes per bench.
3. Precipitation buffer B (ppt B): 300 µl per tube; 2 tubes per bench.
4. PBS (1X): 50 ml per tube; 1 tube per student.

Equipment needed:

1. Big weigh boats with paper towels: 1 per bench.
2. Big glass beaker (1 L): one per bench.
3. Small glass beaker (100 ml) with gauze pad: 2 per bench.
4. Pipetman P1000 and tips: 2 per bench.
5. Pipetman P200 and tips: 2 per bench.
6. Transfer pipettes: 2 per bench.
7. 1.5ml Eppendorf tube: 2 tubes per student.
8. Table microfuge: one per bench.
9. Eppendorf centrifuge: one per 18 students.
10. Vortex: two per lab
    

II. Using Purified IgY To Detect Proteins

Reagents needed:

1. Nitrocellulose membrane blotted with proteins: one membrane per student; it is in the small weigh boat.
2. Blocking solution (PBS [1X]+3%BSA): 5 ml per tube; 1 tube per student. (Store at -20° C between uses.)
3. AP conjugated goat anti-IgY in PBS (1:2000): 4 ml per tube; 1 tube per student; stored at 4^oC until they are ready to use.
4. Western blue: 750µl per tube; 1 tube per student; at RT no light.
5. dd H₂0: 100ml in a flask; 1 flask per bench.

Additional equipment needed:

1. Rocker: 1 per lab
2. Forceps: 1 per bench.

• Background
• General Immunology Information for Teachers
• Protocol
• Biotechnology Field Trips Home Page

Contact: Barbara Bielec, 608-273-9737