Bio-Rad Apoptosis Assays - Manual (Page 15)

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Fig. 3. Preparing a threefold dilution series with a single reconstituted standard.
5. Prepare Samples
The kit has sufficient reagents to run 19 fractionated samples (cytosolic
and nuclear + mitochondrial fractions) in duplicate or 38 total homogenates
in duplicate.
The following reagents are recommended for the preparation of
fractionated samples and total homogenate:
n
Protease inhibitors — complete, Mini; Roche Diagnostics GmbH
(cat #11 836 153 001), or equivalent
n
Refrigerated centrifuge
n
Tubes
a. 1.5 ml conical microcentrifuge tubes, USA Scientific (cat #1415-2500)
b. 2 ml screw cap tubes, USA Scientific (cat #1420-8700)
n
Homogenizer
a. Pestle grinder — Kontes grinder (cat #KT749540-0000), pestle
(cat #KT749520-0090), and 1.5 ml Kontes grinder snap cap tube
(cat #KT749520-0090), or equivalent
Reconstituted
Standard
100 100 100 100 100 100 100 100
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Standard Diluent, µl
150 50 50 50 50 50 50 50 Transfer Volume, µl