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- W2382024800 abstract "Objective To establish an indirect solid phase ELISA for the detection of IgG and IgM class au- toantibodies directed against prothrombin.Methods High activated polyvinylchloride plates were coated with 100μl of 10μg/ml purified human prothrombin in Tris buffered saline(TBS)overnight at 4℃.To avoid non specific bind- ing,wells were blocked with 150μl goat serum.After washing with TBS containing 0.1% Tween 20,100μl of ser- um samples diluted with TBS containing 1% bovine serum albumin(TBS 1% BSA)in 1:100 were added in dupli- cate.Plates were incubated for 1h at room temperature and after washing with TBS Tween,100μl of horse radish peroxidase(HRP)-conjugated goat antihuman IgG or IgM antibodies in TBS 1% BSA were added in the appropriate dilution.After half an hour incubation at room temperature followed by 3 washes,100μl tetramethylbenzidine (TMB)/H_2O_2 were added for 15 min,the reaction was terminated with 50μl 1 mmol/L HCl.Results were read at a wavelength of 450 nm against a background of 650 nm a plate reader.The sensitivity,specificity and reproducibility of ELISA,and its correlation to the imported kit from Germany were studied.Results The method for detecting IgG and IgM,aPT showed good sensitivity,specificity,reproducibility,and stabilization.Conclusion The ELISA kit is successfully developed,it is suitable to use in experimental and clinical application." @default.
- W2382024800 created "2016-06-24" @default.
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- W2382024800 date "2007-01-01" @default.
- W2382024800 modified "2023-09-23" @default.
- W2382024800 title "Detection of serum antiprothrombin antibody by indirect ELLSA" @default.
- W2382024800 hasPublicationYear "2007" @default.
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