Lmportant considerations – Bio-Rad Bio-Plex Pro™ Magnetic Cell Signaling Assays User Manual
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lmportant Considerations
Assay Procedures
n
Please pay close attention to vortexing, shaking, and incubation
times and to Bio-Plex
®
reader PMT (RP1) setting, as these have been
optimized specifically for the cell signaling assays
n
For optimal performance, use only reagents specific for Bio-Plex Pro
™
cell signaling assays. Reagents in other Bio-Plex assay panels have not
been validated for use in the cell signaling assays
n
Do not reuse diluted (1x) coupled beads, detection antibodies, or
streptavidin-PE
n
Wash as outlined in Table 5. Incomplete washes may cause
assay variation
n
If the data are not acquired immediately, the assay plate
may be stored
at 4°C for up to 24 hr protected from light
Assay Quick Guide
Each assay kit comes complete with a printed Bio-Plex Pro Assay Quick Guide
(bulletin 10024930), which can be used to prepare and run a full 1 x 96-well
assay plate. Users can also download a copy at www.bio-rad.com/bio-plex.
Bead Regions and Multiplexing Compatibility
n
Bead regions for all analytes are listed in Table 12
n
Compatible singleplex assays may be mixed to create a multiplex assay
n
Do not mix phosphoprotein assays with corresponding total target
assays (for example, phospho-Akt and total Akt)
Instruments and Software
The Bio-Plex Pro cell signaling assays described in this manual are
compatible with all currently available Luminex-based life science
research instruments. Assays can be read and analyzed with either
Bio-Plex Manager
™
software or Luminex xPONENT software (Section 5,
Read the Plate, under Detailed Instructions).