Model Number: HBsAg ELISA01 Brand Name: Egens Key Specifications/Special Features: This HBsAg ELISA is a qualitative enzyme immunoassay for the in vitro detection of Hepatitis B SurfaceAntigen(HBsAg) in human serum or plasma. It is intended for screening of blood donors and diagnosing patientsrelated to infection with HBsAg.Assay procedure:1. Prepare Reagents: Dilute 1 volume of Concentrated Washing Buffer (20×)with 19 volumes of distilled water,mix well.2. Add Samples and Conjugate: Open the foil pouch and remove the Microplate. Set up 1 well as Blank, 2 wellsas negative control, 2 wells as positive control. After dispensing 50L Samples and 50L Negative Control orPositive Control to the respective wells, add 50μl of Conjugate into each well except the blank. Gentlyvibrating the plate. 3. Incubate: Cover the Microplate with Plate Cover and incubate the Microplate in a thermostat-controlled
water-bath or microplate incubator at 37℃ for 30 minutes.
4. Wash the Plate: Remove the plate cover. Aspirate the contents of all wells. Fill the wells with the diluted
washing buffer ( 10~20 seconds to soak) then aspirate again. Repeat the procedure for 5 times. Make sure
that the rest volume is minimal, by tapping plate onto absorbent paper.
5. Add Substrate: Add 50L of Substrate Solution A and 50L of Substrate Solution B to each well, mix well.
Cover and incubate at 37℃ for 10 minutes.
6. Stop reaction: Add 50L Stop Solution to each well, mix well.
7. Read the absorbance at 450 nm. If a dual wavelength measurement is used, the reference wavelength should
be selected from 620nm to 690nm.