Prepare stock solution
Prepare a B/B Homodimerizer stock solution by dissolving the compound in 100% ethanol. We recommend beginning with a stock of 62.5 mg/ml, which is concentrated enough to allow administration of doses of up to 10 mg/kg (in a dose volume of 4 ml/kg) while keeping the final ethanol concentration at 4%.
In the likely event that the initial dose-response study indicates that doses significantly less than 10 mg/kg can be used routinely, make a less-concentrated stock solution by further dilution in 100% ethanol.
To prepare a 62.5 mg/ml stock solution:
- Add 16 μl of 100% ethanol per 1 mg B/B Homodimerizer.
- Vortex until completely dissolved (let the solution sit for a few minutes before vortexing).
This stock solution can be stored, long-term, at –20°C.
Prepare dosing solution
Prepare the B/B Homodimerizer dosing solution within 30 min of use. Only prepare the volume needed for a given experiment in order to conserve your stock solution. To administer 10 mg/kg in a dose volume of 4 ml/kg, prepare a 2.5 mg/ml dosing solution as follows:
Per 1 ml of dosing solution:
- Pipette 40 μl of 62.5 mg/ml stock solution into a fresh vial.
- Add 100 μl PEG-400 (100%). Vortex.
- Add 860 μl Tween (2%) in water. Vortex.
- Inject within 30 min.
NOTE: To prepare a less-concentrated dosing solution, simply dilute the stock solution to the appropriate degree in 100% ethanol and follow steps 1–4 above. This will keep the final ethanol concentration constant at 4%.
Deliver the B/B Homodimerizer
Deliver B/B Homodimerizer to mice by IP or intravenous (IV) injection of the appropriate volume. For example, to deliver a 10 mg/kg dose, administer 4 ml/kg of the 2.5 mg/ml dosing solution (e.g., 100 μl/25 g).