User`s Manual
Contents
1. N BioChain TET A PE A PEEN Tel 1 888 762 2568 Fax 1 510 783 5386 Email info biochain com User s Manual and Instructions ELISA Kit for IgM Antibody to Hepatitis A Virus Catalog No KO31008096 NAME AND INTENDED USE ELISA Kit for IgM Antibody to Hepatitis A Virus is an in vitro enzyme immunoassay for the detection of HAV IgM in human serum or plasma PRINCIPLE This kit uses capture ELISA method to detect anti HAV IgM The purified rabbit ant human IgM monoconal antibody Anti u chain is coated on the solid phase of multi wells Serum sample Horseradish peroxidase labeled HAVAg are added to coated wells After incubation if HAV IgM is presentin the sample a complex of Anti p chain HAV IgM HAVAg labeled wth HRP will form Wash wells to remove other unbounded serum components incubate with substrate TMB to fom a colored product and measure the absorbance at 450 nm to indicate the presence or absence of HAV IgM in the sample The test is special sensitive reproducible and easy to operate MATERIALS PROVIDED 1 Anti u chain Coated Microwell Plate 1 block 96wells 2 Enzyme Conjugant HAVAg HRP 1 bottle 12ml 3 Negative Control Serum 1 vial 1ml 4 Positive Control Serum 1 vial 1ml 5 Wash Buffer 1 20 dilution prior to use 1 botte 60m 6 Substrate A 1 botte 6ml 7 Substrate B 1 botte 6ml 8 Stop Solution 1 botte 6ml 9 Plastic Bag 1 bag 10 Seal Paper 3 pieces 11 Manual 1 each2. SAMPLE COLLECTION AND PRESERVATION Blood serum samples are routinely prepared form vein Blood plasma sample are routinely prepared with routine amount of anticoagulant such as heparin or sodium ctrate Sample can be stored at 4 C if tested within five days Sample can be stored at 20 C atleast for 3 months Avoid hemolysis and repetitive freeze and thaw of samples Samples with coud or precipitation should be centnfugated or filtered before test Prevent serum from bacteria contamination during collection and storage TES T PROCEDURE 1 Bring ELISA Kit for IgM Antibody to Hepatitis A Virus all reagents and samples to room temperature before use approximately 30 minutes 2 Dilute concentrated wash buffer 1 20 wth ddH2O 3 For each test set one blank two positive and three negative controls Add 50 ul positive and negative control serum into positive and negative control wells respectively without sample diluent 4 Add 50 ul test serum into test wells 5 Cover wells with seal paper incubate for 30 minutes at 37 C 6 Discard the liquid in all wells and fill the wells with wash soluton F 753 3UM RevA KO31008096UA 800ul per well Lay aside for 15 seconds discard the liquid in all wells and fill the wells with wash solution Repeat 5 times and dry wells after last wash 7 Add 100 ul Enzyme Conjugant in each well except the blank well 8 Cover wells with seal paper incubate for 30 minutes at 37 C
3. 9 Wash 5 times the same as step 6 10 Add 50 ul substrate A and B respectively to each well mix gently protected from light and lay aside for 15 minutes at 37 C 11 Add one drop of stop solution 50 ul into each well to stop the reaction induding blank well 12 Measure the absorbance at 450 nm against the blank or measure the absorbance at 450 nm 630 690 nm INTERPRE TATION OF RESULTS Colorimetric Method Cut Off Value calculation COV 0 07 the average OD of negative controls Positive ODssoof sample COV Negative OD oof sample COV 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 PRECAUTIONS 1 The samples should be fresh avoid hemolysis bacteria growing and repetitive freeze and thaw 2 Do notinterchange reagents between Kit lots The seal paper can t be used repeatedly 3 Mix reagents well before use If crystal form in certain reagents such as wash buffer it can be used without problems after warm up and mix well 4 Follow instruction exactly during assay especially in temperature and time for reactions All pipetting devices should be used with care and calibrated regulary following the manufacturers instructions 5 Put the remained reagents to the sealed pouch and return to 2 8 Cin time 6 To prevent cross contamination wear gloves and working suits throughout the proc
4. edure and execute the disinfection and isolation regulations strictly Dispose of all samples and matenals used to perfom the test The 5 0g L liquid sodium hypochlorite solution or 121 C high pressure steam may be used to disinfect samples and materials before disposal The positive control serum in the kit has been inactivated already PACKAGE SIZE 96 tests Kit STORAGE AND STABILITY Store the kitat 2 8 CC EXPIRATION The shelf life is 12 months from the receiving date This Kit is for Research Use Only Active Date 09172012
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