Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 9)

7
Prepare samples, reconstitute lyophilized reagents,
dilute assay buffer to 1x, prepare standards
Add 10 µl blocking buffer to all wells
Add 30 μl standards, blank, samples,
and controls to appropriate wells
Add 10 μl 1x capture beads per well.
Shake at 850 ± 50 rpm for 1 hr at RT
Do not aspirate after incubation
Add 40 μl reconstituted detection antibodies.
Shake at 850 ± 50 rpm for 1 hr at RT
Do not aspirate after incubation
Add 20 μl 1x streptavidin-PE.
Shake at 850 ± 50 rpm for 30 min at RT
Wash 3x with 100 µl assay buffer (1x)
Resuspend beads in 100 μl assay buffer (1x).
Shake at 850 ± 50 rpm for 30 sec
Read plate on Bio-Plex system
Assay Workflow