Enzyme immunoassay for the determination of IgG antibodies to nuclear and cytoplasmic antigensin human serum and plasma
品牌
ANAscreen
样品类型
Serum, Plasma
Analytical Method
Semi-Quantitative
特异性
Frequency distribution of ANA in ANAscreen: 97unselected human sera from healthy donors weretested. All sera were found negative. Thiscorresponds to a diagnostic specificity of 100%.
产品特性
ANAscreenallows the detection of total autoantibodiesto nuclear and cytoplasmic antigens in one sample as asummary parameter in the diagnosis of systemicautoimmunedisorders.Antigeniccombinationofcomplete HeLa nuclei with recombinant proteins andpurified native antigens guarantees a maximum ofsensitivy and specificity for the ANA detection.
组件
Microtiter plate Concentrated wash buffer Sample diluent Conjugate Substrate Stop solution Positive Control Cut-off Control Negative Control
试剂未包括
micropipettes multi-channel pipette or multi-pipettetrough for multi-channel pipette 8-channel wash comb with vacuum pump and wastebottle or microplate washer distilled or de-ionized water glassware
Optimal working dilution should be determined by the investigator.
实验流程
The ANAscreen ELISA Assay Kit is an enzymeimmunoassay for the semi-quantitative determination of IgG antibodies to nuclear and cytoplasmic antigens. Antibodies of the controls and diluted patient samples react with nuclear and cytoplasmic antigens immobilized on the solid phase of microtiter plates. Use of complete HeLa nuclei enriched with recombinant and native antigens guarantees the specific binding of autoimmune antibodies of the specimen under investigation. Following an incubation period of 60 min at room temperature (RT,18...25°C),unbound sample components are removed by a wash step. The bound IgG antibodies react specifically with anti-human-IgG conjugated to horseradish peroxidase (HRP). Within the incubation period of 30 min at RT, excessive conjugate is separated from the solid-phase immunecomplexes by the following wash step. HRP converts the colorless substrate solution of 3,3’,5,5’-tetramethylbenzidine (TMB) added into a blue product. The enzyme reaction is stopped by dispensing an acidic solution into the wells after 15 min at room temperature turning the solution from blue to yellow. The optical density (OD) of the solution at 450 nm is directly proportional to the amount of specific antibodies bound. Patient ratios are calculated by dividing the respective OD of the specimen with the calculated cut-off OD.
限制
仅限研究用
储存条件
4 °C
储存方法
Upon receipt, all components of the ANAscreen have tobe kept at 2 - 8 °C, preferably in the original kit box.
Systemic autoimmune diseases such as systemic lupuserythematosus,scleroderma,rheumatoidarthritis,Sjögren’s syndrome, dermatomyositis, mixed connectivetissue disease are characterized by the appearance of avariety of autoantibodies directed against componentsof the cell nucleus.Although significance and pathological relevance ofsome auto-antibodies are not completely revealed yet,the detection of auto-antibodies is widely establishedand plays an important role in the diagnosis of systemicautoimmune diseases.