RADIM Ihr Spezialist für In-vitro-Diagnostik (IVD)

ALLERgen Total-IgE
ELISA kit

Enzymimmunoassay for the quantitative determination of total immunoglobulin E (IgE) in human serum or plasma

The ALLERgen Total-IgE assay is an immunoassay based on the sandwich principle. The patient sample is pipetted into the wells coated with a monoclonal anti-IgE antibody. During the first incubation, this antibody binds the IgE of the sample. Upon addition and subsequent incubation with an anti-IgE-biotin conjugate, a “sandwiched” anti-IgE: IgE: anti-IgE-biotin complex is formed. The further incubation steps with a streptavidin-peroxidase-conjugate and the substrate lead to the development of a blue coloration. The enzymatic reaction is then blocked with a stopper liquid and yellowing occurs. The resulting color is measured photometrically at 450 nm. The color intensity is directly proportional to the IgE concentration in the sample.

Contents and components: Total-IgE Kit for 96 Tests

IgE microtiter plate:
The kit contains a microtiter plate with 12 strips x 8 wells. Each Well is coated with monoclonal anti-Human IgE (mouse). Store the unused strips together with the desiccant at 2-8°C in the closeable plastic bag.
Anti-IgE-Biotin-Conjugat:
The vial contains 13 ml anti-IgE biotin conjugate in TRIS buffer, pH 8.1, with stabilisers and preservatives. Ready to use.
Total-IgE-calibration:
6 vials (0.5 ml each) of human IgE in concentrations 0 – 10 – 50 – 250 – 500 – 1000 kIU/l (2nd IRP 75/502 WHO), in horse serum with 0.01% ProClin 300 as a preservative. Ready to use.
Incubation buffer:
The vial contains 18 ml TRIS buffer, pH 8.4, with 0.1% TWEEN® 20 and 0.005% ProClin 300 as preservative. Ready to use.
Sample dilution liquid:
The vial contains 10.0 ml of horse serum with 0.01% ProClin 300 as a preservative. Ready to use.
HRP-Streptavidin-Conjugate:
The vial contains 13 ml of streptavidin-peroxidase-conjugate in red colored buffer, pH 5.5, with 0.001% ProClin 300 as a preservative. Ready to use.
Substrate HS:
The vial contains 13 ml of a stabilized mixture of 3.3′, 5.5′ Tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2). Ready to use.
Stopping Liquid:
The vial contains 13 ml of 0.3 M sulphuric acid. Ready to use.
Washing buffer concentrate 10x:
The vial contains 100 ml of a 10x concentrated washing liquid, with 2.5 μg of amphotericin B. To prepare the working liquid, mix the contents of the vial with 900 ml of distilled water.