Run the assay, Considerations – Bio-Rad Human Metabolic and Hormone Assays User Manual
Page 20
18
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