Direct Saliva Melatonin ELISA

Enzyme immunoassay for highly sensitive, quantitative determination of melatonin in human saliva.

INTENDED USE:The NovoLytiX Direct Saliva Melatonin ELISA (EK-DSM) is intended for highly sensitive, quantitative determination of melatonin in human saliva. For research use only. Not intended for use in diagnostic procedures.

GENERAL INFORMATION:The NovoLytiX Direct Saliva Melatonin ELISA is a competitive immunoassay using a capture antibody (Ab) technique. The polyclonal Kennaway G280 anti-melatonin antibody (5, 6) has been coated onto the microtiter plate, provided in the kit. During the 16-20-hours overnight incubation melatonin present in the pre-treated saliva and controls as well as in the calibrators compete with biotinylated melatonin for the binding sites of this highly specific antibody. After washing, the enzyme label, streptavidin conjugated to horseradish peroxidase (HRP), is added, which binds to the melatonin-biotin-antibody complexes captured on the coated wells during a 60-minutes incubation step. Unbound enzyme label is then removed by a second washing step and TMB substrate (tetramethylbenzidine) is added to the wells. In a further 30-minutes incubation step, a chromophore is formed in inverse proportion to the amount of melatonin present in the sample. The color turns from blue to yellow after the addition of an acidic stop solution and can be measured at 450 nm.

KIT CHARACTERISTICS:
- Method: ELISA
- Tests: 96
- Incubation Time / Conditions: 1 min (shaker), 18 h (2-8°C), 1 h, 30 min (RT, shaker)
- Standard Range: 0.5 - 50 pg/ml
- Sensitivity analytical: 0.47 pg/ml (LoD)
- Final Sample Volume: 100 µl
- Sample Type: saliva
- Sample Pretreatment: Pretreatment solution
- Isotope / Substrate: TMB 450 nm
- Internal Controls: 2
- Regulatory Status: RUO