Bio-Rad Human Metabolic and Hormone 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.
Reagent Volume, µl Reagent Volume, ml
Standards Mix 150 Blocking Buffer 1.5
Control 1 100 Standard Diluent 1.0
Control 2 100 Detection antibiotics 4.8
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 ambient/room temperature (RT).
a. Mix by inversion to ensure all salts are in solution.
b. Prepare 1x assay buffer — dilute 1 part 10x assay buffer (60 ml) with
9 parts of dH
2
0 (540 ml).
B. Dilution of Standard (1:3 Serial Dilution)
1. Label 9 polypropylene tubes S1 through S8 and Blank.
2. Transfer the reconstituted standard into the tube labeled S1.
Bio-Plex Pro ™ RBM Metabolic and
Hormone 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.
9. Add 20 µl of diluted SA-PE to the required plate wells.
SA-PE Dilution Volume of SA-PE, µl Volume of 1x Assay
Buffer, µl
Total Volume, µl
1:10 225 2,025 2,250
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 1x 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 RBM Metabolic and Hormone Assays Quick Guide
For Use with
Instruction Manual #
Bio-Plex Pro ™ RBM Metabolic and Hormone Assays 10041818
The Bio-Plex suspension array system includes fluorescently labeled microspheres and
instrumentation licensed to Bio-Rad Laboratories, Inc. by Luminex Corporation. MAGPIX is a
trademark of Luminex Corporation.
Bio-Plex Pro RBM kits are manufactured by Myriad RBM. Myriad RBM is a trademark of Myriad
RBM, Inc.
Life Science
Group
Sig 1213 10041817 Rev B US/EG
Bio-Rad
Laboratories, Inc.
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