User`s Manual
Contents
1. 9 gt Biochain www biochain com Tel 1 888 762 2568 Fax 1 510 783 5386 Email info biochain com User s Manual and Instructions ELISA Kit for Hepatitis Be Antigen Catalog No KO31006096 INTENDED USE ELISA Kit for Hepatitis B e Antigen is an in vitro enzyme immunoassay for the detection of HBeAg in human serum or plasma PRINCIPLE The purified Anti HBe is coated on the solid phase of multi wells Serum sample and Horseradish peroxidase labeled with Anti HBe conjugated are added to coated wells After incubation if HBeAg is present in the sample a complex of Anti HBe HBeAg Anti HBe labeled with HRP will form Wash wells to remove other unbounded serum componenis incubate with substrates TMB to form a colored product and measure the absorbance at 450nm to indicate the presence or absence of HBeAg in the sample The test is specific sensitive reproducible and easy to operate STORAGE AND STABILITY Store the kit at 2 8 C The kit is stable unopened for 12 months upon receiving it MATERIALS PROVIDED 1 Anti HBe Coated Microwell Plate 2 Enzyme Conjugant 1 bottle 6 5 ml 3 Positive Control Serum 1 vial 1 ml 4 Negative Control Serum 1 vial 1 ml 5 Wash Buffer 1 20 dilution prior to use 1 bottle 50 ml 6 Substrate A 1 bottle 8 5 ml 7 Substrate B 1 bottle 8 5 ml 8 Stop Solution 1 bottle 8 5 ml 9 Seal Paper 2 pieces PRECAUTIONS 1 The samples should be fresh and avoid hemolysis2. This Kit is for Research Use Only Active Date 05202015
3. bacterial growth and repetitive freeze thaw cycle 2 Do not interchange reagents between different lots 3 The seal paper CANNOT be used repeatedly 4 Mix reagents well before use If crystal form in certain reagents such as wash buffer warm bottle vials to redissolve Mix well 5 Follow the instruction provided in the manual especially for incubation temperature and reaction time All pipetting devices should be used with care and calibrated regularly following the manufacturer s instructions 6 Put the remaining reagents in the sealed pouch and return them to 2 8 C for short period of storage 7 To prevent cross contamination wear gloves and lab coats throughout the procedure and disinfection spills immediately Dispose of all samples and materials used to perform the test To disinfect used samples and materials before disposal one should autoclave at 121 C or use 5 0 g L liquid sodium hypochlorite solution The positive control serum in the kit has been inactivated already F 753 3UMRevA 1 block 96wells KO31006096UB ASSAY PROCEDURE 1 For each test set one blank two positive and two negative controls add 0 05ml serum sample positive and negative control serum into the coated wells then add one drop approximately 0 05ml of enzyme conjugant into the same coated wells The blank well is omitted mix thoroughly cover wells with seal paper and incubate for 30 60 minutes at 37 C 2 Manual W
4. ash Procedure Discard the liquid in the coated wells and bring them to dry Fill the wells with 300ul wash buffer discard the liquid Repeat 5 times and then bring them to dry Automatic Wash Procedure Select the automatic operations of washing 5 times and bring them to dry after the operation 3 Add one drop approximately 0 05ml of substrate A and B respectively to each well seal the plate with seal paper mix thoroughly and incubate for 10 15 minutes at 37 C Avoid exposure to light 4 Add one drop approximately 0 05ml of stop solution into each well mix thoroughly to terminate the reaction Measure the absorbance at 450nm against the blank or measure the absorbance at 450nm 630 690 nm INTERPRETATION OF RESULTS Colorimetric Method Cut Off Value calculation COV the average OD of negative controls x 2 1 Positive OD s50 of sample gt COV Negative OD 5 0 of sample lt COV Invalid If the average OD of negative controls 20 1 the result is invalid In this event repeat the test If problem persists contact the local distributor Notes If the absorbance of negative controls is below 0 05 calculate it as 0 05 If the absorbance of negative controls is above 0 05 calculate it as its original value PERFORMANCE CHARACTERISTICS Sensitivity 2NCU ml the Biological Reference Reagents of Chinese Clinical Test Center OD 21 500 Specificity the average OD of 20 normal negative samples lt 0 030 Precision CV lt 15
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