1.4: 1x400ulM3986-01E: Positive Control, Low 1.0-2.1: 1x400ulM3986-01F: Serm Diluent: 1x30mlM3986-01G: IgG, IgA, IgM (HRP) Goat x human: 1x15mlM3986-01H: Wash Buffer, 20x: 1x50mlM3986-01J: Tetramethylbenzidine (TMB): 1x15mlM3986-01K: Stop Solution: 1x15mlStorage and Stability:Store all components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.">
Systematic autoimmune disease is characterized by the presence of circulating autoantibodies directed to a wide variety of cellular antigens. Systemic lupus erythematosis (SLE), commonly referred to as Lupus is the best known of these diseases. Other possible connective tissue diseases include mixed connective tissue disease (MCTD), Sjogren syndrome, sclerodema, and polymyositis/dermatomyositis.Mitochondria (M2) antigen is part of the pyruvate dehydrogenase complex. Approximately 90-95% of patients with primary biliary cirrhosis (PBC) have antibodies to M2. Antimitochondria antibodies occur occasionally in other liver conditions and scleroderma. The antibody is rarely seen in other conditions. Classically, antibodies to autoimmune antigens are detected by double immunodiffusion. However, the test is lengthy and suffers weak sensitivity. ELISAs combine greater sensitivity with ease of use. Many ELISAs have been developed and validated for detecting autoantibodies to various antigens.Intended Use:Mitochondria, BioAssay ELISA Kit is intended for the detection and semi-quantitation of antibodies to mitochondria in patient serum. The assay is to be used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of primary biliary cirrhosis. Principle of the Assay:The United States Biological Mitochondria, BioAssay ELISA Kit is used to detect IgG, IgA, and IgM antibodies to Mitochondria antigens. Purified Mitochondria antigens are attached to a solid phase microassay well. Enzyme-Linked Immunosorbent Assays (ELISA) rely on the ability of biological materials (i.e., antigens) to adsorb to plastic surfaces such as polystyrene (solid phase). When antigens bound to the solid phase are brought into contact with a patient's serum, antigen specific antibody, if present, will bind to the antigen on the solid phase forming antigen-antibody complexes. Excess antibody is removed by washing. This is followed by he addition of an IgG, IgA, IgM (HRP) goat anti-human which then binds to the antibody-antigen complexes. The excess conjugate is removed by washing, followed by the addition of Chromogen/Substrate, tetramethylbenzidine (TMB). If specific antibody to the antigen is present in the sample, a blue color develops. When the enzymatic reaction is stopped with 1N H2SO4, the contents of the wells turn yellow. The color, which is indicative of the concentration of antibody in the serum, can be read on a suitable spectrophotometer or ELISA microwell plate reader. Kit Components:M3986-01A: Microtiter Plate: 1x96 wellsM3986-01B: Calibrator: ≥0.3 1x400ulM3986-01C: Negative Control 0.0-0.8: 1x400ulM3986-01D: Positive Control, High >1.4: 1x400ulM3986-01E: Positive Control, Low 1.0-2.1: 1x400ulM3986-01F: Serm Diluent: 1x30mlM3986-01G: IgG, IgA, IgM (HRP) Goat x human: 1x15mlM3986-01H: Wash Buffer, 20x: 1x50mlM3986-01J: Tetramethylbenzidine (TMB): 1x15mlM3986-01K: Stop Solution: 1x15mlStorage and Stability:Store all components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.