Human 6-Acetylmorphine (6-MAM) ELISA Kit

Sample

Urine, whole blood, serum, oral fluid

Species Reactivity

Human

Detection Method

cELISA

Intended Use

For the determination of trace quantities of 6-MAM in human urine, whole blood, serum and oral fluid.

Contents of Kit

1. 6-MAM Dilution Buffer, 30 mL (ready-to-use). Phosphate buffer solution provided to dilute samples. Note: Specific for 6-MAM assay.
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. 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, 14 mL (ready-to-use). Drug-horseradish peroxidase conjugate. Do not dilute.
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. Acid Stop Solution, 14 mL (ready-to-use). 1N H₂SO₄ used to stop the enzyme reaction.
7. Qualitative QC Positive Control, 750 μL provided (diluted in synthetic matrix). Do not dilute.
8. Qualitative QC Negative Control, 750 μL provided (diluted in synthetic matrix). Do not dilute.

Storage

This kit can be used until the expiration date on the label when stored refrigerated at 2-8°C. Note: Some kits require controls to be stored frozen immediately upon receipt. Reference kit label for details.

Sensitivity

For the I-50 in 6-AM Dilution Buffer, the value is 3 ng/mL. When testing Whole Blood with a 1:10 dilution (1 part sample to 9 parts 6-AM Dilution Buffer), the I-50 is 40 ng/mL. In Serum, using a 1:5 dilution (1 part sample to 4 parts 6-AM Dilution Buffer), the I-50 is 10 ng/mL. For Urine with a 1:10 dilution (1 part sample to 9 parts 6-AM Dilution Buffer), the I-50 is 40 ng/mL. Finally, in Oral Fluid with a 1:4 dilution (1 part sample to 3 parts 6-AM Dilution Buffer), the I-50 is 10 ng/mL.
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 6-AM Dilution Buffer. The drug concentration that shows 50% less color activity than the zero standard is considered to be the I-50.

General Description

The 6-MAM ELISA (Enzyme-Linked ImmunoSorbent Assay) test 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).

Standard Curve

The typical standard curve is generated with the drug in 6-AM Dilution Buffer, plotting % B/B0 (y-axis) against 6-Acetylmorphine concentration in ng/mL (x-axis, approximately 0.1-100 ng/mL). The extent of color development is inversely proportional to the amount of drug in the sample or control. The drug concentration that shows 50% less color activity than the zero standard is considered to be the I-50. (Source manual provides the curve as a graph only, without an OD data table.)