Purpose
This kit is used for the determination the existence of Human Tetanus Antibody
(TAB) in human serum, plasma, cell culture supernatants, and other biological
fluids.
Principle of the assay
The kit assays human TAB
in the samples by the method of purified Tetanus antigen pre-coated
enzyme-labeled SPA ELISA. The solid-phase antigen is prepared by purified human
Tetanus antigen which is pre-coated onto a microplate. After loading of samples,
the TAB in the controls or samples bind with the coated antigen. Then the
Staphylococcal protein A (SPA)-enzyme immunoglobulin 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 TAB
can be calculated according to OD from positive or negative controls.
Calculation
1.
Average the duplicate
readings for each control and sample and subtract the
average of blank well (1 x sample buffer only).
2.
Determine P/N value: P/N= ODsample
/ODnegative. If ODnegative is less than 0.10, please
use 0.10 as the value. ODpositive or ODnegative is the OD450
from positive or negative control.
3.
If P/N ≥ 2.1, the testing
sample is TAB positive; if P/N < 1.5, the testing sample is TAB negative. If 1.5
≤P/N<2.1, samples are suspected, may need repeating test or verified by other
methods.
4.
The normal range of ODnegative
should be less than 0.21, and ODpositive should be more than 0.40,
otherwise the experiment may have some errors and maybe need repeating.
5.
If the ODsample is
more than 2.5 or less than 0.1, it is out of the
optimal
test range. Dilution or concentration of sample, or adjust incubating time may
be necessary for accuracy.
Kit Performance
1.
Sample from healthy people should be negative.
2.
Specificity: simultaneous detection of recombinant or natural human TAB without
cross-reaction with other cytokines.
3.
Repeatability: both
inter-assay and intra-assay were less than 10 %.
*Lead time of this kit is 1-2 weeks. Shipped by
FedEx air.
*Storage: 2 - 8 C. Shelf Time: 6 months.
*Kit Experimental Time:< 2 hs
>>Trial pricing is available for first time use for certain customers,
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Important note:
Products are intended for laboratory or
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.