Bio-Rad Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays - Manual (Page 27)

25
20x Detection Detection Antibody
# of Wells Antibodies, µl Diluent, µl Total Volume, µl
96 150 2,850 3,000
48 75 1,425 1,500
20x Detection 20x Detection Detection
Antibodies, µl Antibodies, µl Antibody
# of Wells Singleplex #1 Singleplex #2 Diluent, µl Total Volume, µl
96 150 150 2,700 3,000
48 75 75 1,350 1,500
Table 19. Mixing two singleplex assays or a premixed panel + singleplex assay.
Preparing 1x detection antibodies from 20x stock (includes 25% excess volume)
Table 20. Premixed panel or one singleplex assay.
Table 21. Mixing two singleplex assays or a premixed panel + singleplex assay.
Table 22. Preparing 1x detection antibodies from two stocks at different concentrations.
Mixing human insulin (20x) with human IL-6 (10x) is one example.*
5. After incubating the beads, samples, standards, and blank, slowly
remove and discard the sealing tape.
6. Wash the plate three times with 100 µl wash buffer.
7. Vortex the diluted (1x) detection antibodies gently for 5 sec. Pour
into a reagent reservoir and transfer 25 μl to each well using a
multichannel pipet.
10x Detection 10x Detection Detection
Antibodies, µl Antibodies, µl Antibody
# of Wells Singleplex #1 Singleplex #2 Diluent, µl Total Volume, µl
96 300 300 2,400 3,000
48 150 150 1,200 1,500
* Due to differences in dilution factors, it is not possible to multiplex adiponectin, adipsin,
VCAM-1, or ICAM-1 with other diabetes or cytokine assays.
20x Detection 10x Detection Detection
Antibodies, µl Antibodies, µl Antibody
# of Wells Diabetes Cytokines Diluent, µl Total Volume, µl
96 150 300 2,550 3,000
48 75 150 1,275 1,500