Bio-Rad Human Metabolic and Hormone Assays - Manual (Page 13)

11
Setting Up a Vacuum Manifold
Calibrate the vacuum manifold by placing a standard 96-well flat bottom
plate on the unit and adjusting the pressure to –1 to –3” Hg. In general,
100 µl liquid should take 3–4 sec to clear the well. For more detailed
instructions, refer to bulletin #10005042.
4. Prepare Reagents
1. Reconstitute the following lyophilized reagents in dH
2
0 before use
according to the table below.
a. Allow vial to sit at room temperature for a minimum of 5 min, not to
exceed 30 min.
b. Mix by vortexing at a medium setting.
2. Bring the 10x assay buffer to room temperature (RT).
a. Mix by inversion to ensure all salts are in solution.
b. Prepare 1x assay buffer — dilute 1 part 10x assay buffer (60 ml) with
9 parts of dH
2
0 (540 ml).
Reagent dH
2
O Volume
Standards mix 150 µl
Control 1 100 µl
Control 2 100 µl
Blocking buffer 1.5 ml
Standard diluent 1.0 ml
Detection antibodies 4.8 ml
Table 4. Reagent volume.