ELISA HBsAb test

Model Number: ELISA HBsAb Key Specifications/Special Features: Instructions for use HBsAb ELISA QuantitativeAssay procedure1. 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, 12 wellsas Calibration Curve Standards (run the standards in duplicates), 0 mIU/ml, 10 mIU/ml, 20 mIU/ml, 40mIU/ml, 80 mIU/ml, 160 mIU/ml. After dispensing 50L Samples and 50L Calibration Curve Standards tothe respective wells, add 50μl of Conjugate into each well except the blank . Gently vibrating the plate.3. Incubate: Cover the Microplate with Plate Cover and incubate the Microplate in a thermostat-controlledwater-bath or microplate incubator at 37℃ for 60 minutes.4. Wash the Plate: Remove the plate cover. Aspirate the contents of all wells. Fill the wells with the dilutedwashing buffer ( 10~20 seconds to soak) then aspirate again. Repeat the procedure for 5 times. Make surethat 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.
Results
1. Plot the absorbance (log-OD) for each duplicate calibration standard on the Y (logarithmic ordinate) versus
the corresponding anti-HBs concentration (log-mIU/ml) on the X (logarithmic abscissa) on double-logarithmic
paper (do not average the duplicates of the calibration standards before plotting).
2. Draw the standard curve through the plotted points (best-fit).
3. To determine the concentration of anti-HBs for an unknown, locate the absorbance (OD) for each unknown on
the Y-axis of the graph, find the intersecting point on the standard curve, and read the concentration
(log-mIU/ml) from the X-axis of the graph. Calculate the concentration of the unknown in mIU/ml.
Precautions
1. Allow all kit components to reach room temperature before use.
2. Follow the direction insert to control the reaction temperature and time strictly.
3. Do not mix components of different lot numbers to use.
4. The kit should be store at 2-8℃. Do not use kit components beyond their expiration date.