Bio-Rad Bio-Plex Pro™ RBM Canine Kidney Toxicity Assays - Manual

This guide can be used to prepare and run a full 1 x 96-well assay plate.
For more information on a given step, refer to the corresponding section
of the complete instruction manual. New users can download the manual,
which includes detailed instructions and a list of kit components, at
www.bio-rad.com/bio-plex.
A. Reagent Preparation
1. Reconstitute the following lyophilized reagents in dH
2
0 before use
according to the table below.
a. Allow vial to sit at room temperature for a minimum of 5 min, not to
exceed 30 min.
b. Mix by vortexing at a medium setting.
2. Bring the 10x assay buffer to room temperature (RT).
a. Mix by inversion to ensure all salts are into solution.
b. Prepare 1x assay buffer — dilute 1 part 10x assay buffer with
9 parts of dH
2
0.
Bio-Plex Pro ™ RBM Kidney Toxicity Assays
Quick Guide
IMPORTANT! Pay close attention to vortexing, shaking, and incubation instructions.
Deviation from the protocol may result in low assay signal and assay variability.
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 ™ ).
Bio-Plex Pro Assay Quick Guide
For Use With
Instruction Manual #
Bio-Plex Pro ™ RBM Human, Rat, and Canine Kidney Toxicity Assays 10028258
Volume of Volume of
SA-PE Dilution SA-PE, µl 1x Assay Buffer, µl Total Volume, µl
1:10 225 2,025 2,250
Reagent Volume, µl Reagent Volume, ml
Standards mix 150 Blocking buffer 1.5
Control 1 100 Standard diluent 1.0
Control 2 100 Detection antibodies 4.8
Life Science
Group
Sig 1212 10028259 Rev B
Bio-Rad
Laboratories, Inc.
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