Human hepatitis B virus surface antigen,HBsAg ELISA Kit , 96T
is used for the determination the existence of Human Hepatitis B Virus Surface
Antigen (HBsAg) in human serum, plasma, cell culture supernatants, and other
Principle of the assay
assays human HBsAg
the samples by the method of purified HBs antibody pre-coated enzyme-labeled
ELISA. The solid-phase antigen is prepared by purified human HBs antibody which
is pre-coated onto a microplate. After loading of samples, the HBsAg in the
controls or samples bind with the coated antibody. Then the
HBs antibody-enzyme labeled conjugates has been added into wells. After washing
to remove any unbound reagent, the tetramethylbenzidine (TMB) substrate is
added. TMB substrate produces blue color when catalyzing by the HRP enzyme.
After reaching the desired color intensity, the reaction is terminated by
addition of an acidic Stop solution which changes the solution color from blue
to yellow. Determining the absorbance (OD Value) at wave length 450 nm by the
microwell reader, and the existence of HBsAg
can be calculated according to OD from positive or negative controls.
Average the duplicate readings for each control and sample and subtract the
average of blank well.
Determine P/N value: P/N= ODsample / (0.1 + ODnegative).
If ODpositive is more than 2.5, please use 2.5 as the value. ODpositive
or ODnegative is the OD450 from positive or negative
P/N ≥ 1.0, the testing sample is HBsAg positive; if P/N< 1.0, the testing sample
is HBsAg negative.
normal range of ODnegative should be less than 0.1, and ODpositive
should be more than 1.0, otherwise the experiment may have some errors and
maybe need repeating.
the ODsample is more than 2.5 or less than 0.1, it is out of the
test range. Dilution or concentration of sample may be necessary for accuracy.
1. Sample from healthy people should be negative, e.g. P/N < 1.0.
Specificity: simultaneous detection of recombinant or natural human HBsAg
without cross-reaction with other cytokines.
inter-assay and intra-assay were less than 10 %.
*Lead time of this kit is 1-2 weeks. Shipped by
*Storage: 2 - 8 C. Shelf Time: 6 months.
*Kit Experimental Time:< 2 hs
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manufacture use and are not to be used for diagnosis or any other purpose, which
include but are not limited to, unauthorized commercial uses, ex vivo or in
vivo therapeutic uses. Not for Human or Animal Consumption.