Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 20)

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Panel Sample
Dilution
Volume of
Sample, µl
Volume of
Extraction
Buffer
Volume of
Neutralization
Buffer
Volume of Sample
Dilution Buffer 2
IGF 1:30 20 180 µl Mix 50 µl extracted
sample with
50 µl neutralization
buffer Mix the neutralized
sample with
50 µl sample dilution
buffer 2
IGFBP 1:20 10 N/A N/A 190 µl
Note: Sample dilution factor for culture media samples must be optimized
by the end user.
6. Run the Assay
Considerations
n
Bring all assay components and samples to room temperature before use
n
Use calibrated pipets and pipet carefully, avoiding bubbles. Use new
pipet tips for every volume transfer
n
Pay close attention to vortexing, shaking, and incubation instructions.
Deviation from protocol may result in low assay signal and assay variability
n
Assay incubations are carried out in the dark on a shaker at 850 ± 50 rpm.
Cover the plate with a plate seal and protect from light with aluminum foil
Table 8. Summary of wash options and protocols. After each assay step, select the
appropriate Bio-Plex Pro wash station program or perform the appropriate manual wash
step as summarized below.
Table 7. Serum and plasma sample dilution guidelines.
Bio-Plex Pro or Bio-Plex Pro II Handheld Magnet or
Pro II Wash Station Wash Station Vacuum Manifold
Assay Step Magnetic Program Vacuum Program Manual Wash Steps
After SA-PE incubation MAG x3 VAC x3 3 x 100 μl