| Antigen-antibody interaction can result in immune complex (IC) formation and it is body’s normal and continuous immunological function. The antigens may be tissue fixed or free in serum and other body fluids. ICs may form locally in the extravascular fluids or systematically in the circulation. The ICs are removed harmlessly by reticuloendothelial system. However, large deposition of ICs in vascular structures with subsequent activation of inflammatory mediator pathways such as the complement (C) sequence leads to an “ICs” disease state accompanied by tissue damage. The fate of ICs depends upon many variables, including the nature of antigen and antibodies (class of antibodies), the degree of lattice formation (ICs size), the rate of production of ICs and the functional integrity of phagocytic system. |
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| Catalog # | C5090 |
| Circulating immune complexes (CIC) related abnormalities include anaphylatoxin production, cell lysis, leukocyte stimulation, and activation of macrophages and various other cells. Fixation of CICs to cell membranes, vessel walls can induce tissue damage as in some cases of glomerulitis. Elevated levels of CIC have been implicated in many diseased including autoimmune diseases (SLE and RA), neoplastic diseases (Hodgkin’s, leukemia), infectious disease due to bacteria (subacute bacterial endocarditis, SBE; leprosy), viruses (hepatitis, mononucleosis), parasites (malaria, schistomiasis), and many other unclassified disorders. |
| Because of pathogenic role of CIC in many diseases, considerable research has been directed towards the development of specific, sensitive and reliable techniques for measuring CIC. Currently, a large number of methods are available to detect and quantitate CIC. These techniques takes advantage of the specific physicochemical properties of CIC and/or of their ability to bind to the surface of cells (platelets, Raji cells, and macrophages) or molecules (C1q, monoclonal proteins with RF activity, polyclonal RF, or the agglutinating factor found in mouse serum). However, none of these assays may be completely satisfactory in all clinical situations. Because of this WHO recommended in 1978 that at least two different methods be used to detect CIC adequately. |
| Animals models of disease (natural or induced; spontaneous lupus like disease) provide valuable insights into human disease and allow well defined controlled experiments that are not possible in humans. |
| CIC ELISA kit has been specially developed to measure CIC in mouse serum. |
| ELISA Kit Features |
| Precoated, ready to use 96-well strip plate for multiple uses over 12–18 months. Direct sample 100ul analysis. No sample dilution/processing allows rapid analysis. 75 min. assay time, three convenient room temperature incubation (30+30+15 min). Time saving ready-to-use substrate solution and other assay components. High sensitivity, excellent precesion. Up to 1 year shelf life. |
| This kit is for in vitro research use only. |
| Principle of the Test | The CIC ELISA test is based on the binding of CIC (complement fixing type) to C1q coated on microtiter plates. Diluted serum samples are incubated with the coated plates. After a washing step, anti-mouse IgG(HRP) conjugate is added. After another washing step, chromogenic substrate (TMB) is added and color developed. The enzymatic reaction (color) is directly proportional to the amount of CIC present in the sample. The reaction is terminated by adding stopping solution. Absorbance is read at 450nm. Concentration of CIC in samples can be determined by comparing with positive controls or standards. To confirm the presence of CIC, serum samples can be diluted in regular sample diluent and in “Confirmation Solution” (provided in the kit) that is known to disrupt binding of CIC to C1q. Both samples are then measured at the same time. A decrease of CIC by at least 30% in “Confirmation Solution” confirms the presence of CIC in samples. |
| Kit Components | C5090A: Microtiter Plate, 1x96 wells (12 strips). Coated with purified C1q. |
| C5090B: Negative Control (10X), 1x150ul. |
| C5090C: Positive Control (10X), 1x150ul. |
| C5090D: Sample Diluent (10X), 1x10ml. Supplied as a red-colored liquid. Dilute 1:10 with distilled water. |
| C5090E. IgG (HRP) (100X), 1x200ul. Dilute 1:100 with 1X Sample Diluent before use. |
| C5090F: Confirmatory Solution, 1x10ml. Supplied as a green-colored liquid. |
| C5090G. Wash Buffer (100X), 1x10ml. Dilute 1:100 with distilled water before use. |
| C5090H: TMB Substrate, 1x12ml. |
| C5090J: Stop Solution, 1x12ml. |
| Storage and Stability | All kit components are stable at 4°C. The whole kit stability is 6 months. The unused strips should be stored tightly covered with adhesive film and with the desiccant in the bag. |
| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
