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

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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