Radioactive antiglobulin testing with 125I anti-C4 and anti-C3. 1978

D E Jenkins, and R M Johnson, and W H Moore

The report describes methods for quantitating C4 and C3 coating of red blood cells using 125I anti-C4 and anti-C3. Two methods of preparing antisera were used; one involving absorption and elution from complement coated cells, and the other employing IgG fractions of antisera prepared on DEAE cellulose. White-cell-free red blood cells were used in the test procedure. Both the labeled antisera and the test cells were diluted in cold carrier proteins, initially bovine serum albumin, but more recently normal rabbit serum. We have found that normal rabbit serum is particularly effective in reducing the high uptake of nonspecific counts on normal cells, which had previously been a major technical problem for the radioactive antiglobulin technique. These methods are highly reproducible and relatively easy to perform once the labeled antisera are prepared. Furthermore, the use of labeled antisera permits analysis of specificity by the highly sensitive technique of radioimmunoelectrophoresis. One problem with the technique is that results indicate uptake of labeled antisera, rather than amounts of coating substances. Nonetheless, the ease of performance and the reproducibility of the method, make the procedure very useful in comparative studies of uptake of C4 and C3 by red blood cells.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D007457 Iodine Radioisotopes Unstable isotopes of iodine that decay or disintegrate emitting radiation. I atoms with atomic weights 117-139, except I 127, are radioactive iodine isotopes. Radioisotopes, Iodine
D002352 Carrier Proteins Proteins that bind or transport specific substances in the blood, within the cell, or across cell membranes. Binding Proteins,Carrier Protein,Transport Protein,Transport Proteins,Binding Protein,Protein, Carrier,Proteins, Carrier
D003176 Complement C3 A glycoprotein that is central in both the classical and the alternative pathway of COMPLEMENT ACTIVATION. C3 can be cleaved into COMPLEMENT C3A and COMPLEMENT C3B, spontaneously at low level or by C3 CONVERTASE at high level. The smaller fragment C3a is an ANAPHYLATOXIN and mediator of local inflammatory process. The larger fragment C3b binds with C3 convertase to form C5 convertase. C3 Complement,C3 Precursor,Complement 3,Complement C3 Precursor,Complement Component 3,Precursor-Complement 3,Pro-C3,Pro-Complement 3,C3 Precursor, Complement,C3, Complement,Complement, C3,Component 3, Complement,Precursor Complement 3,Precursor, C3,Precursor, Complement C3,Pro C3,Pro Complement 3
D003181 Complement C4 A glycoprotein that is important in the activation of CLASSICAL COMPLEMENT PATHWAY. C4 is cleaved by the activated COMPLEMENT C1S into COMPLEMENT C4A and COMPLEMENT C4B. C4 Complement,C4 Complement Component,Complement 4,Complement C4, Precursor,Complement Component 4,Pro-C4,Pro-complement 4,C4, Complement,Complement Component, C4,Complement, C4,Component 4, Complement,Component, C4 Complement,Pro C4,Pro complement 4
D003298 Coombs Test A test to detect non-agglutinating ANTIBODIES against ERYTHROCYTES by use of anti-antibodies (the Coombs' reagent.) The direct test is applied to freshly drawn blood to detect antibody bound to circulating red cells. The indirect test is applied to serum to detect the presence of antibodies that can bind to red blood cells. Anti-Human Globulin Consumption Test,Antiglobulin Consumption Test,Antiglobulin Test,Coombs' Test,Antihuman Globulin Consumption Test,Direct Antiglobulin Test,Direct Coombs Test,Indirect Antiglobulin Test,Indirect Coombs Test,Anti Human Globulin Consumption Test,Antiglobulin Consumption Tests,Antiglobulin Test, Direct,Antiglobulin Test, Indirect,Antiglobulin Tests,Antiglobulin Tests, Direct,Antiglobulin Tests, Indirect,Consumption Test, Antiglobulin,Consumption Tests, Antiglobulin,Coomb Test,Coomb's Test,Coombs Test, Direct,Coombs Test, Indirect,Direct Antiglobulin Tests,Indirect Antiglobulin Tests,Test, Antiglobulin,Test, Antiglobulin Consumption,Test, Coombs,Test, Coombs',Test, Direct Antiglobulin,Test, Direct Coombs,Test, Indirect Antiglobulin,Test, Indirect Coombs,Tests, Antiglobulin,Tests, Antiglobulin Consumption,Tests, Direct Antiglobulin,Tests, Indirect Antiglobulin
D004912 Erythrocytes Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN. Blood Cells, Red,Blood Corpuscles, Red,Red Blood Cells,Red Blood Corpuscles,Blood Cell, Red,Blood Corpuscle, Red,Erythrocyte,Red Blood Cell,Red Blood Corpuscle
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000906 Antibodies Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody

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