Intended Use:This kit is designed for in vitro quantification of histamine in various biological fluids by competitive direct enzyme-linked immunosorbent assay (CD-ELISA). This kit is intended for use in investigative biomedical research only. It is not for human clinical diagnostic use. This Histamine BioAssay™ ELISA Kit should not be used for determining histamine levels in scombroid fish.
Principle of the Assay:This Histamine BioAssay™ ELISA Kit is a competitive direct ELISA in a microwell format that allows users to obtain exact concentrations of histamine in nanograms per milliliter. The microwells in this assay kit are pre-coated with a monoclonal antibody to histamine. The sample or standard solution is first added to the antibody coated microplate. Next, the enzyme conjugate is added and the mixture is shaken and incubated at RT for 45 minutes. During the incubation, unbound (free) histamine in the samples or standards is allowed to compete with enzyme HRP-labeled histamine (conjugate) for antibody binding sites. The plate is then washed, removing all the unbound material. The bound enzyme conjugate is detected by the addition of a one-component substrate which generates color by horseradish peroxidase. An optimal color is generated after 30 minutes. A microplate reader is then used to take an absorbance reading at 650nm.
Quantitative test results may be obtained by measuring and comparing the absorbance reading of the sample wells against the standard curve using a log-logit curve fitting model. The extent of color development is inversely proportional to the amount of histamine in the sample or standard. For example, the absence of histamine in the sample will result in a bright blue color, whereas the presence of histamine will result in decreased or no color development.
Range: 2.5-50.0ng/ml
Intra-assay Precision: ≤10%
Inter-assay Precision: ≤10%
Kit Components:H5085-01: Wash Buffer (25X), 1x30mlH5085-02: TMB Substrate, 1x20mlH5085-03: PBS Sample Diluent, 1x1LiterH5085-04: Histamine Enzyme (HRP), 1x6mlH5085-05: Standard (0ng/ml), 1x500ulH5085-06: Standard (2.5ng/ml), 1x500ulH5085-07: Standard (5ng/ml), 1x500ulH5085-08: Standard (10ng/ml), 1x500ulH5085-09: Standard (20ng/ml), 1x500ulH5085-10: Standard (50ng/ml), 1x500ulH5085-11: Microtiter Plate, 1x96 wells
Storage and Stability:Store all components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
