Methods in Enzymology Vol.93 Immunochemical Techniques, Part F: Conventional Antibodies

This chapter discusses the intra-lymph node technique used to inoculate rabbits with small quantities of antigen. In the experiment, 0.5 ml of an isotonic saline solution containing 2% (w/v) Evans blue dye between two toes of each hind foot of unanesthetized, 2–3kg female rabbits was injected. This can be done for at least 1 hr or up to 18 hr before inoculation. By this process, the dye becomes concentrated in the lymph nodes and makes them much easier to find. Sufficient antigen–Freund's adjuvant was made so that the popliteal lymph node of each hind leg can receive 0.1 ml of emulsion containing between 2 and 100 μg of antigen. An antigen dose producing a favorable response can be determined only by trial and error. The bleeding and boosting program depends on the use of antigen, the similarity of structure of the antigen with other peptides that affect specificity, and the availability of antigen. A variety of antigens in the 20,000–22,000 molecular weight range, as well as a 15 amino acid peptide coupled to bovine serum albumin, have been inoculated successfully by this procedure.

• Series: Methods in Enzymology (Book 93)
• Hardcover: 480 pages
• Publisher: Academic Press;
• Language: English
• ISBN-10: 0121819930
• ISBN-13: 978-0121819934
• Product Dimensions: 6 x 1 x 9 inches

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