B. dilution of standard (1:3 serial dilution), C. sample preparation – Bio-Rad Apoptosis Assays User Manual

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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).

4. Prepare working standards (S2–S8) by serial dilution. Transfer the

appropriate volume of standard into each of the labeled tubes with
standard diluent as outlined above.

5. Vortex each standard at a medium setting before proceeding with the next

serial dilution. Change pipet tip at each dilution step.

C. Sample Preparation

1. Refer to instruction manual #10033631 for detailed sample preparation.

Note: Pay close attention to cell lysis, homogenization, and fractionation

protocols in the manual. Analytes of interest may be localized to the
cytosolic fraction or to the nuclear and mitochondrial fraction.

2. Prepare sample dilutions in 0.5 ml or 1.0 ml polypropylene tubes as

required for the assay.

Bio-Plex Pro Assay Quick Guide

Standard

Volume of Standard Diluent, µl

Volume of Standard, µl

S2

100

50 of S1

S3

100

50 of S2

S4

100

50 of S3

S5

100

50 of S4

S6

100

50 of S5

S7

100

50 of S6

S8

100

50 of S7

Blank

100

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