Bio-Rad Rat Diabetes Assays - Manual (Page 26)

24
Prepare and Add Detection Antibodies
Instructions are provided for diluting the detection antibodies to a 1x
concentration. When mixing diabetes and cytokine assays, keep in mind
the stock concentrations of detection antibodies as shown below.
Table 14. Stock concentration of detection antibodies.
1. While the samples are incubating, use Tables 15–17 to calculate the
volume of detection antibodies and detection antibody diluent needed.
Detection antibodies should be prepared 10 min before use.
2. Add the required volume of Bio-Plex detection antibody diluent to a
15 ml polypropylene tube.
3. Vortex the stock detection antibodies for 15–20 sec at medium
speed, then perform a 30 sec spin to collect the entire volume at the
bottom of the tube.
4. Dilute detection antibodies to 1x by pipetting the required volume into
the 15 ml tube.
Each well of the assay requires either 1.25 μl (20x stock) or 2.5 µl
(10x stock) adjusted to a final volume of 25 μl in detection
antibody diluent.
Stock Concentration of
Assay Detection Antibodies
Human/NHP, mouse, and rat diabetes 20x
Human and mouse cytokines (groups I, II) 10x
Rat cytokines (group I) 20x
Mouse cytokines (group III) 20X