Human Herceptin ELISA Kit

Sample

Serum, EDTA plasma

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Human Herceptin/trastuzumab (Humanized Anti-HER-2/neu ) ELISA Kit is an enzyme linked immunosorbent assay intended for the quantitative measurement of Herceptin levels in human serum or EDTA plasma.

Contents of Kit

1. HER-2 Coated Microplate
One microplate with twelve by eight strips (96 wells total) coated with recombinant HER-2 protein. The plate is framed and sealed in a foil zipper bag with a desiccant.
2. Herceptin Tracer Antibody
One vial containing 12 mL concentrated Tracer Antibody in a stabilized protein matrix. This reagent should be stored at 2-8°C and is stable until the expiration date stated over the kit box.
3. ELISA Wash Concentrate
One bottle containing 30 mL of 30-fold concentrate. Before use the contents must be diluted with 870 mL of demineralized water and mixed well. Upon dilution, this yields a working wash solution containing a surfactant in phosphate buffered saline with a non-azide, non-mercury preservative. The diluted wash solution may be stored at room temperature and is stable until the expiration date stated over the kit box.
4. ELISA HRP Substrate
One bottle containing 12 mL of tetramethylbenzidine (TMB) with hydrogen peroxide. This reagent should be stored at 2-8°C and is stable until the expiration date stated over the kit box.
5. ELISA Stop Solution
One bottle containing 12 mL of 1N sulfuric acid. This reagent may be stored at 2 – 8 °C or at room temperature and is stable until the expiration date stated over the kit box.
6. Herceptin Calibrators
Six vials containing ready-to-use Herceptin in a serum based matrix with a non-azide preservative. Refer to the vial for exact concentration of the calibrator. These calibrators should be stored at 2-8°C, (-20°C for longer storage).
7. Herceptin Controls
Two vials containing ready-to-use Herceptin in a serum based matrix with non-azide preservative. Refer to the vial for the acceptable concentration ranges for the controls. These controls should be stored at 2 – 8 °C, (-20°C for longer storage).
8. Herceptin Concentrated Assay Buffer
One bottle containing 30 mL of 4-fold concentrated buffer matrix with protein stabilizers and preservative. This reagent should be stored at 2 – 8 °C and is stable until the expiration date on the kit box. Before use the concentrated buffer must be diluted with 90 mL of demineralized water and mixed well. Upon dilution, this yields as 1× Assay Buffer, which serves as the patient sample diluent containing a surfactant in phosphate-buffered saline with a non-azide preservative. This reagent should be stored at 2-8°C and is stable until the expiration date stated over the kit box.

Storage

The kit must be stored at 2 – 8°C upon receipt. All components are stable until expiration date stated over the label on kit box. Reagents from different kit lot numbers should not be combined or interchanged. Prior to use, allow all reagents to come to room temperature.

Precision

The intra-assay precision was validated by measuring three spiked samples with 16 replicate determinations.

The inter-assay reproducibility was validated by measuring two control levels in duplicate in 16 individual assays.

Sensitivity

The sensitivity (lowest limit of detection) of this Herceptin ELISA as determined by the corresponding absorbance value of 2-fold calibrator deviation above the mean on 16 determinations of zero calibrator is 0.245 ng/mL.

General Description

Herceptin (Trastuzumab) is a humanized monoclonal antibody used in cancer treatment for Human Epidermal Growth Factor Receptor-2 (HER-2) over-expressing tumors. In the case of these tumors, Herceptin binds to and inhibits tumor growth. Herceptin can also be labeled with various toxins to enhance tumor suppression. Studies have also found that Herceptin targets gastric cancer stem cells characterized by CD90 phenotype.

Standard Curve

EXAMPLE DATA AND CALIBRATOR CURVE
A typical absorbance data and the resulting calibrator curve from this Herceptin ELISA are represented. This curve should not be used in lieu of calibrator curve generated with each assay.