Hemolytic assay for goat (caprine) and swine (porcine) complement |
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Authors: | Oyekan P P Barta O |
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Institution: | Department of Veterinary Microbiology and Parasitology, Clinical Immunology Research Unit, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana 70803, USA. |
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Abstract: | Optimal conditions for assaying hemolytic complement of goat (caprine) and swine (porcine) sera were determined. Effects of the following were tested: pH, ionic strength, calcium and magnesium ion concentrations, time and temperature of incubation, and ethylenediamine tetracetate concentration. Guinea pig erythrocytes sensitized with goat or cattle antibodies were the most sensitive target cells for goat complement. Sheep and cattle erythrocytes sensitized with rabbit hemolysin were the best target cells for swine complement. Barbital buffer, pH 7.3, ionic strength of 90 nmM relative salt concentration, containing 0.5 mM CaCl2 and 1 mM MgCl2 was the best for swine complement assay. Goat complement lysed best in a barbital buffer, pH 8, ionic strength of 90 to 120 mM of relative salt concentration, in presence of 0.5 mM CaCl2 and 1 mM MgCl2. The optimal incubation temperature was 37 degrees C for both complements. The complement dependent lysis required 75 minutes to reach its maximum. Ethylenediamine tetracetate in 4 mM concentration completely inhibited lysis by both species complements. The CH50 for goat sera varied between 18 and 75 per ml, in swine sera between 75 and 210 per ml. |
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Keywords: | A antibody BB barbital - NaCl buffer C hemolytic complement [The abbreviations for complement (C) and complement components (C1 through C9) used in this paper conform to the recommendations outlined in “Nomenclature of Complement” Bull World Health Org 39 (1968): 935–938] CH50 E erythrocytes EA erythrocytes sensitized with an optimal dose of antibody GlBB glucose-barbital-NaCl buffer mS millisiemens = millimho = a unit of conductivity RSC relative salt concentration i e molarity of NaCl solution equivalent to the given conductivity |
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