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This finished, useful and basic guide is designed to be a realistic development of experimental protocols that an green investigator might persist with whilst embarking on a biochemical or biotechnology venture.
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Tissue-culture cell lines (European Collection of Cell cultures). 3. Minimal essential medium, Eagles’ (Cat. 32360-026), fetal bovine serum (Cat. 10084-069), L-glutamine 200 mM (Cat. 25030-024), Fungizone (Cat. 152-018), penicillin/streptomycin (Cat. 15140-114), trypsin-EDTA (x1, Cat. 45300-019), all from Gibco, Gaithersburg, MD. 4. MEM nonessential amino acids (100x; Cat. 11140-035, Gibco) and Insulin-transferrin-selenium-G. 5. Supplement (Cat. 41400-045; Gibco) are required for the CaCO-2 cell-line growth.
The inoculation mixture should be prepared aseptically to minimize contamination with endotoxins. In addition, the immunogen itself may have innate adjuvant activity and this should be checked before preparing complex adjuvant formulations. Conjugation of the immunogen to a carrier may impart innate adjuvant activity (see Note 5). Because of necessity, many of these tests require the use of living animals to obtain standard values for the various tests, however, some attempts could be made to extrapolate from data in published records of adjuvant research.
1988) Comparisons of two in vitro cytotoxicity assays—the neutral red (NR) and tetrazolium MTT tests. Toxicol. in Vitro 2, 1–6. 18. British Standards Institution (1983) Evaluation of medical devices for biological hazards. Part 7. Method of test for skin irritation of extracts from medical devices. British Standard 5736, 7. , 2, Park Street, London, W1A 2BS, England, pp. 1–3. 19. Kensil, C. , and Marciani, D. (1991) Structure and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex.