Sample
Urine, Blood, Oral Fluid, Forensic Matrices
Species Reactivity
Human, Research Animal
Intended Use
For the determination of trace quantities of Flunitrazepam and/or other metabolites in human urine, blood, oral fluid.
Contents of Kit
1. EIA Buffer: 40 mL (ready-to-use). Phosphate buffered saline solution with bovine serum and a preservative. Provided for dilution of drug-enzyme conjugate concentrate and samples.
2. Wash Buffer Concentrate (10X): 20 mL. Phosphate buffered saline solution with a surfactant. Dilute 10 fold with deionized or ultrapure water before use. Diluted wash buffer is used to wash all unbound conjugate and samples from the plate after the conjugate incubation.
3. K-Blue Substrate: 20 mL (ready-to-use). Stabilized 3,3', 5,5' Tetramethylbenzidine (TMB) plus Hydrogen Peroxide (H2O2) in a single bottle. It is used to develop the color in the wells after washing. Light sensitive.
4. Drug-Enzyme Conjugate: 200 μL. Drug-horseradish peroxidase concentrate. Dilute 180X before use.
5. Antibody Coated Plate: A 96 well Costar plate, in strips of 8 break-away wells, coated with anti-drug antiserum. The plate is ready for use as is. Do not wash.
6. Red Stop Solution: 20 mL (ready-to-use). Non-acidic reagent used to stop the enzyme reaction.
7. Qualitative QC Positive Control: 750 μL provided (synthetic human urine). Do not dilute.
8. Qualitative QC Negative Control: 750 μL provided (synthetic human urine). Do not dilute.
Storage
This kit can be used until the expiration date on the label when stored refrigerated at 2-8°C. Store controls frozen if not used within 10 days. Note: Some kits require controls to be stored frozen immediately upon receipt. Reference kit label for details.
Sensitivity

The term I-50 is used to define the sensitivity of the test. This number is derived from a standard curve generated with the drug in EIA Buffer. The drug concentration that shows 50% less color activity than the zero standard is considered to be the I-50.
General Description
The Flunitrazepam ELISA Kit is a qualitative one-step kit designed for use as a screening device for the detection of drugs and/or their metabolites. The kit was designed for screening purposes and is intended for forensic use only. It is recommended that all suspect samples be confirmed by a quantitative method such as gas chromatography/mass spectrometry (GC/MS).
Citations
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PARK, DG; DEBLAS, AL; et al. PEPTIDE HETEROGENEITY OF GABA-A BENZODIAZEPINE RECEPTORS IN BOVINE CEREBRAL-CORTEX AND CEREBELLUM. BRAIN RESEARCH 550:279-286(1991).
Feigenspan, A; Gustincich, S; et al. Pharmacology of GABA(A) receptors of retinal dopaminergic neurons. JOURNAL OF NEUROPHYSIOLOGY 84:1697-1707(2000).