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Bio-plex pro assay quick guide 5 – Bio-Rad Bio-Plex Pro™ TGF-β Assays User Manual

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6. Reconstitute a single vial of standards in 500 µl of a diluent similar to the

final sample type or matrix as shown below. Vortex for 5 sec and incubate
on ice for 30 min.

7. Prepare a fourfold standard dilution series and blank as shown below.

Vortex for 5 sec between liquid transfers.

8. After thawing samples, activate by adding 1 volume of 1 N HCI to

5 volumes of sample. Vortex, incubate at RT for 10 min. Neutralize by
adding the same volume of base (1.2 N NaOH/0.5 M HEPES). After
treatment, dilute samples as shown below.

9. Vortex the 20x coupled beads for 30 sec and dilute to 1x in Bio-Plex

assay buffer as shown below. Protect from light.

# of Wells

20x Beads, µl

Assay Buffer, µl

Total Volume, µl

96

288

5,472

5,760

Bio-Plex Pro Assay Quick Guide 5

Reconstituted

Standard

72 150 150 150 150 150 150 150 150

S1 S2 S3 S4 S5 S6 S7 S8 Blank

Diluent, µl

128 50 50 50 50 50 50 50

Transfer Volume, µl

Sample Type

Diluent for Standard

Add BSA

Serum and plasma

Standard/sample diluent mix (1:3)

None

Culture media, with serum

Culture media

None

Culture media, serum-free

Culture media

To 0.5% final (w/v)

Lavage, lysate, other fluids

Sample diluent

To 0.5% final (w/v)

Sample Type

Diluent

Add BSA

Sample Dilution

Serum and plasma

Sample diluent

None

1:16 final*

Culture media, with serum

Culture media

None

User optimized

Culture media, serum-free

Culture media

To 0.5% final

User optimized

Lavage, lysate, other fluids

Sample diluent

To 0.5% final

User optimized

* For example, activate 25 μl sample, neutralize, and bring to a final volume of 400 μl.