Bio-Rad Apoptosis Assays - Manual (Page 22)

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7. Cover and incubate at 850 ± 50 rpm, as in Step 4, for 1 hr at RT. Do
not aspirate after incubation.
8. Prepare the required dilution of streptavidin-PE (SA-PE) as outlined in
Table 8.
Note: Volumes in the table are for an entire 96-well plate. Smaller
volumes can be prepared, provided that dilution ratios are maintained.
9. Add 20 µl of diluted SA-PE to the required plate wells.
10. Cover and incubate at 850 ± 50 rpm, as in Step 4, for 30 min at RT.
11. Wash the plate three times with 100 µl 1x assay buffer.
12. After the final wash, resuspend the beads in 100 µl assay buffer. Cover
plate as in Step 4 and shake the plate at 850 ± 50 rpm for 30 sec.
13. Remove the plate seal and read plate at low PMT (Bio-Plex ® 200),
standard PMT (Bio-Plex 3D), or default settings (Bio-Plex ® MAGPIX ™ ).
7. Read Plate
Bio-Plex Manager ™ software is recommended for all Bio-Plex Pro ™ assay
data acquisition and analysis. Instructions for Luminex xPONENT software
are also included. For instructions on using other xMAP system software
packages, contact Bio-Rad Technical Support or your regional Bio-Rad
field applications specialist.
Volume of Volume of
SA-PE Dilution SA-PE, µl 1x Assay Buffer, µl Total Volume, µl
1:10 225 2,025 2,250
Table 8. SA-PE dilution.