The detection method in microarray chips is based on microscopy analysis via the use of fluorophore or chemoluminescence
labels that determine abundance of nucleic acid sequences.
In Abstacts of the 11th International Symposium on Bioluminescence and Chemoluminescence
After washing three times (10 minutes each) with decreasing concentrations of SDS (0.5%, 0.1%, and 0%) in 2x SSPE, the membranes were incubated for 2 minutes with the enhanced chemoluminescence
(ECL) detection reagents (Amersham Life Science, s`Hertogenbosch, the Netherlands), and then were exposed to an ECL hyperfilm (Amersham Life Science) for 30 minutes and overnight to visualize the bound probe.
Alkaline phosphatase-conjugated goat antimouse IgG (Dianova) was used at a dilution of 1:20 000 for the detection of specific antibody binding with a chemoluminescence
kit (Tropix) and an exposure time of 10 min.
produced by DNA-RNA hybrids is measured in the Gen-Probe luminometer.
A horseradish peroxidase-conjugated second antibody was added and the membrane was incubated 1 h, followed by ECL chemoluminescence
Antibody binding was detected with a horseradish peroxidase-linked goat anti-rabbit or horse anti-mouse IgG antibody (Cell Signaling #7074 and #7076, resp.; 1:1000-1: 2000 dilution in TBS-Tween/5% milk) incubated for 1 h at room temperature and enhanced chemoluminescence
(ECL Western blotting analysis system, GE Healthcare/Amersham-Biosciences, Freiburg, Germany) was detected by film autography.
We observed that the chemoluminescence
induced by the peroxyl radical generation, initiated by AAPH in the ORAC assay, decreased following addition of lichen extracts; ORAC values were 0.77 [micro]mol TE/mg sample for Vc and 3.06 [micro] mol TE/mg sample for Ci, indicating the lichens' different capacities for scavenging peroxyl radicals.
Anti-diphtheria and anti-tetanus, anti-PT and anti-FHA antibody titres were also measured by ELISA; anti-HBsAg titres were measured by VITROS chemoluminescence
(ECi)/ECiQ Immunodiagnostic System; and anti-poliovirus type 1, 2 and 3 antibody titres (1/dil) were measured by microneutralisation.
The antigen-antibody complexes were visualized using the chemoluminescence
substrate SuperSignal West Pico (Pierce, Rockford, IL) as recommended by the manufacturer.
Although two cumbersome methods for measuring urinary TGF-[beta]1 include a bioassay (26) and an ELISA method using an enhanced chemoluminescence
system with a detection range of 0.1-2.5 [micro]g/L (27), the current method is simpler and more reliable.