Bio-Plex Pro Human Immunotherapy Panel Quick Guide
6.
Prepare a fourfold standard dilution series and blank as shown.
Vortex
at medium speed for
5 sec
between liquid transfers.
Note:
Standards are at S1 concentration after reconsititution and the
controls are ready to use after reconstitution. Controls are included with
the fixed panel only.
50
250
50
50
50
50
50
50
Reconstituted
Standard
150
0
150
150
150
150
150
150
150
S1
S2
S3
S4
S5
S6
S7
S8
Blank
Standard Diluent, µl
Transfer Volume, µl
Standard Serial Dilution
7. Vortex
the coupled beads at medium speed for
30 sec
and
dilute to 1x
in Bio-Plex Assay Buffer as shown. Protect from light.
Premixed Panels
Number of Wells
10x Beads, µl
Assay Buffer, µl
Total Volume, µl
96
570
5,130
5,700
Singleplex Assays
Number of Wells
Singleplex #1
20x Beads, µl
Singleplex #2
20x Beads, µl
Assay Buffer, µl Total Volume, µl
96
285
285
5,130
5,700
Note:
20x singleplex beads allow multiplexing up to 20 analytes.
Running the Assay
1. Vortex
the diluted (1x) beads.
Add 50 µl
to each well of the assay plate.
2. Wash the plate two times
with
100 µl
Bio-Plex Wash Buffer.
3. Vortex
the samples, standards, blank, and control.
Add 50 µl
to each well.
4.
Cover the plate with sealing tape. Incubate on shaker at
850 ± 50 rpm
at
RT for
30 min
.
5.
With 10 min left in the incubation,
vortex
the detection antibodies for
5 sec
and quick-spin to collect liquid.
Dilute to 1x
as shown.
