ELISA (Double Antibody Sandwich Method) - Detection of HBsAg

The carrier is coated with a known antibody (anti-HBs), and then the sample to be tested containing the antigen is added, and after the incubation, the antigen is bound to the coated antibody, and then the enzyme-labeled specific antibody (enzyme-labeled anti-HBs) is added, and the enzyme-labeled antibody is added. The antigen is attached to the antigen, and finally the substrate solution is added, and the antigen content is determined based on the color reaction.

[Material] HBsAg kit, this kit contains:
【method】
1. Add 0.05 ml per well to the specimen to be tested, and set 2 wells of HBsAg positive control, 2 wells of HBsAg negative control, 1 well of blank control, then add 1 drop per well of enzyme conjugate (0.05 ml, blank control well). Mix well and incubate at 37 ° C for 30 minutes.
2, wash the plate: discard the liquid in the reaction strip hole, pat dry, fill each hole with washing liquid, discard the patted dry, patted dry after repeated five times.
3. Color developing agent: first add 1 drop (0.05 ml) per color of the developer A, then add 1 drop (0.05 ml) per hole of the developer B, mix, and incubate at 37 ° C for 10 minutes.
4. Add 1 drop (0.05 ml) of stop solution to each well and mix.
【result】
Colorimetry: Wavelength 450nm, first use the blank hole to zero point, then read the optical density value of each hole.
The OD value of the sample/the average OD value of the negative control ≥ 2.1 was judged to be positive, otherwise it was negative.
Remarks: The average OD value of the negative control is less than 0.05 for 0.05, and the higher than 0.05 is calculated based on the actual OD value. The positive control OD value ≥ 0.8, the experimental results are valid.
【Precautions】
1. Store the kit at 2 °C ~ 8 °C.
2. The reagent should be shaken before use, and discarded 1~2 drops and then added vertically.
3. Take out all the bottled reagents in the kit and the microporous strips required for the specimen to be tested from the refrigerated environment, and equilibrate for 30 minutes at room temperature before testing. The rest should be sealed in the refrigerator for later use.
4. The specimen to be tested is not allowed to be preserved by NaN3.
5, do not use the same batch of reagents.
6. The result judgment must be completed within 10 minutes.
7. If the concentrated washing solution crystallizes, please set it at 37 °C until it dissolves.

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