Owner's manual

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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

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 ambient temperature (RT).

a. Mix by inversion to ensure all salts are into solution.

b. Prepare 1x assay buffer — dilute 1 part 10x assay buffer (60 ml) with

9 parts of dH

0 (540 ml).

Bio-Plex Pro

™

RBM Apoptosis 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.

For Use With Instruction Manual #

Bio-Plex Pro

™

RBM Apoptosis Assays 10033631

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

Summary of content (4 pages)

PAGE 1
Bio-Plex Pro™ RBM Apoptosis Assays Quick Guide For Use With Instruction Manual # Bio-Plex Pro™ RBM Apoptosis Assays 10033631 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.
PAGE 2
Bio-Plex Pro Assay Quick Guide B. Dilution of Standard (1:3 Serial Dilution) 1. Label 8 polypropylene tubes S1 through S8. 2. Transfer the reconstituted standard into the tube labeled “S1.” 3. Add the appropriate amount of the standard diluent into the labeled tubes according to the table below (this will be sufficient for duplicate standard curves and blanks).
PAGE 3
Bio-Plex Pro Assay Quick Guide 3. For sample analysis, bring samples to a final protein concentration of 500 µg/ml by diluting with lysis dilution buffer (LDB). Further dilution may be necessary for targets with high expression levels. 4. Centrifuge samples at 500 x g for 5 min to remove particulates prior to use. Note: Controls are ready to use after reconstitution. No further dilution is needed. D. Dispensing of Reagents 1. Add 10 µl of blocker to all wells of the plate. 2.
PAGE 4
Bio-Plex Pro Assay Quick Guide 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™).