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Assay workflow – Bio-Rad Human Metabolic and Hormone Assays User Manual

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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