Bio-Rad Bio-Plex Pro Human Angiogenesis Reagent and Diluent Kits - Manual (Page 10)

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Section 7
Standard Preparation
Two vials of Iyophilized angiogenesis standards are provided in each Bio-Plex
Pro™ angiogenesis assay. However, only one vial is required per 96-well plate.
The product insert provided with the assay lists the concentration of the
reconstituted standard. This procedure will prepare enough standard to run
each dilution in duplicate.
Reconstitute Standards
1. Gently tap the glass vial containing the lyophilized angiogenesis
standard on a solid surface to ensure the pellet is at the bottom.
2. Reconstitute 1 vial of lyophilized standard with 500 µl of the
standard diluent. Do not use assay buffer to dilute standards.
3. Gently vortex 1–3 sec and incubate on ice for 30 min.
4. Be consistent with the incubation time for optimal assay performance.
Prepare Standard Dilution Series
The angiogenesis concentrations specified for the 8-point standard
dilution set have been selected for optimized curve fitting using the 5-
parameter logistic (5PL) or 4-parameter logistic (4PL) regression in
Bio-Plex Manager™ software. Results generated using dilution points
other than those listed in this manual have not been optimized.
1. Label a set of 1.5 ml Eppendorf tubes as shown in the diagram on
the next page.
2. Pipet the appropriate volume of standard diluent into the tubes (see
diagram on next page). Use serum standard diluent for serum and
plasma samples and culture medium for culture samples.