Lumigen APS-5 utilizes a unique technology for the chemiluminescent detection of alkaline phosphatase conjugates and provides superior sensitivity and ease of use. Reaction of the acridan substrate with an AP label produces sustained high-intensity chemiluminescence. Lumigen APS-5 is ideally suited for solution assays of phosphatase activity and for phosphatase-linked immunoassays.
Lumigen APS-5 in Homogeneous Alkaline Phosphatase Assays
- Excellent sensitivity -- less than 10-19 moles AP can be measured.
- Rapid develpment of peak signal reduces assay time and increases throughput.
- Linear calibration curves with slopes of log-log plots equal to 1.0 -- one order of magnitude more enzyme yields one order of magnitude more luminescence.
- Sustained luminescence -- timing is less critical. Light intensities can be read at any time to produce linear calibration curves.
- Analytical results are insensitive to temperature from 22°C to 35°C reducing the need for precise temperature control.
Rapid Chemiluminescent Immunoassay
A sandwich immunoassay for thyroid stimulating hormone (TSH) performed with an Immulite TSH Third Generation TSH Assay kit yielded a linear plot from 0.003 mIU/L to 75 mIU/L with excellent assay precision. Lumigen APS-5 was substituted for the normal dioxetane substrate, Lumigen PPD.
The image below from a western blot analysis of human transferrin was made after a 10 minute incubation of the PVDF membrane with Lumigen APS-5 with a 1 minute exposure to X-OMAT film. Equivalent results were obtained using nitrocellulose membranes. No other chemiluminescent reagent allows complete interchangeability of blotting membrane.