Bio-Rad Human MMP and TIMP Assays - Manual (Page 2)

7. Reconstitute the vial of standards in 10% serum-based standard diluent
(or a diluent similar to your sample matrix) by adding 781 µl of diluent.
Reconstitute the vial of control in 250 µl of the same diluent, as shown
below. Vortex at medium speed for 5 sec and incubate all vials on ice for
precisely 30 min.
8. Prepare a threefold standard dilution series and blank as shown below.
Vortex at medium speed for 5 sec between liquid transfers.
Note: The controls are ready to use after reconstitution. No dilution is
needed. Controls are included with the fixed panel only.
9. After thawing samples, prepare according to the guidelines shown below.
10. Vortex coupled beads at medium speed 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
150 150 150 150 150 150 150 150
S1 S2 S3 S4 S5 S6 S7 S8 Blank
Diluent, µl
75 75 75 75 75 75 75 Transfer Volume, µl
Reconstituted
Standard
Sample Type Diluent for Standards and Controls* Add BSA
Serum and plasma Sample diluent HB None
Culture media, with serum Culture media None
Culture media, serum-free Culture media To 0.5% final
Lavage, lysate, other fluids Sample diluent HB To 0.5% final
* If using diluents other than the standard diluent HB provided, then users must establish
their own control ranges.
Sample Type Dilution Factor Diluent
Serum and plasma MMP: 1:10 dilution Sample diluent HB
Fluids MMP: 1:4 and 1:40 Diluent + 0.5% BSA w/v
Bio-Plex Pro Human MMP Assays Quick Guide