HYBRIDOMA TECHNIQUES
Spring Quarter, 2007-2008
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Welcome to the homepage for this course. I hope that you will find this page to be useful as you as you prepare each week for Hybridoma Techniques lab. You can download and print each LAB procedure and any DISCUSSION NOTES from the links for each week's labs.
Each LAB procedure and set of DISCUSSION NOTES is also available as a PDF file. This format may be much easier to download and print. . Some links from PDF files do not work. ALL links from HTML files do work. The PDF files will require the Adobe Acrobat Reader to read PDF format. If you don't have a copy on your computer, click here for a free download. To go to links from PDF files, hold down the control key (Ctrl) while clicking on the link.
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Lab
Instructor
Kim Corbett
Offices: 08-1334
Phones: 475-6039
Email: kacsbi@rit.edu
You can
send an email by simply clicking on my name
or my email address above
to open an email
dialogue box already addressed
OR
creating your own email message addressed
to kacsbi@rit.edu
Suggested
Readings
Monoclonal
Antibodies
Monoclonal
Antibodies (different site)
Monoclonal
Antibody Production - The Basics
Uses
for monoclonal antibodies
The
Mouse in Science: Monoclonal Antibodies
Antibodies
and therapy
Monoclonal
Antibody Therapy
Some
monoclonal antibodies that have been introduced into human medicine
Ethical
Issues
Animal
Welfare Act (Animal
Welfare Act - Summary PDF version)
AVMA Guidelines on Euthanasia (American Veterinary Medical Association, 2007)
Acceptable Methods for Euthanasia
of Laboratory Animals at the University of Minnesota
Guide
for the Care and Use of Laboratory Animals
The
American Veterinary Association - Animal Welfare
Comparision
of Ascites vs. Cell Culture for Production of Monoclonal Antibodies
Alternatives
to Ascites Production of Monoclonal Antibodies
Tentative
Grading
Resume
Skills
Tentative Lab Schedule
Week #1 (March 10, 12)
LAB
- Preparation of Lymphocyte Cell Suspension (PDF
version)
LAB
- Myeloma Cell Line Characteristics and Preparation (PDF
version)
DISCUSSION
NOTES - Introduction (PDF
version)
DISCUSSION NOTES - Mouse Myeloma
Cell Line (PDF
version)
Week
#2 (March 17, 19)
LAB - Fusion of Lymphocytes and Myeloma Cells (practice) (PDF version)
DISCUSSION
NOTES - Fusion (PDF
version)
Week
#3 (March 24, 26)
LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)
Week #4 (March 31, April 2)
LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)
DISCUSSION
NOTES - HAT Medium (PDF
version)
DISCUSSION NOTES - 8-Azaguanine
Resistance (PDF
version)
DISCUSSION
NOTES - HT Medium (PDF
version)
Week #6 (April 14, 16)
Quiz #1 (will cover Weeks 1-5)
LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)
Week #7 (April 21, 23)
LAB
- Intraperitoneal Injection (PDF
version) KLH, pristane, LAB - cells (PDF
version)
DISCUSSION
NOTES - Immunization of Mice (PDF
version)
DISCUSSION NOTES - Intraperitoneal
Injection of Mice (PDF
version)
LAB - Intravenous Injection (PDF version) - NOT required, but worth trying
LAB - ELISA for the Detection of anti-KLH Antibodies (screening uncloned hybrids) (PDF version)
DISCUSSION
NOTES - ELISA (PDF
version)
LAB - Cloning by Limiting Dilution (PDF version)
DISCUSSION
NOTES - Cloning by Limiting Dilution (PDF
version)
DISCUSSION NOTES - Feeder Cells (PDF
version)
Week #9 (May 5, 7)
LAB - ELISA for the Detection of anti-KLH Antibodies (screening cloned hybrids) (PDF version)
DISCUSSION NOTES - Pristane-Priming of Mice (PDF version)
Week #10 (May 12, 14)
Quiz #2 (will cover Weeks 6-9)
LAB
- Transferring Cells to 24-well Plates and T-25 Flasks (PDF
version)
LAB -
Collection of ascites tumor cells from pristane-primed mice (demonstration
only) (PDF
version)
DISCUSSION NOTES - Expansion of cell culture volume (PDF version)