ATTAC mouse model dimerizer administration
Reagents
- AP20187, the dimerizer from Ariad Pharmaceuticals. When I receive the dimerizer, I usually prepare solution by adding 100% ethanol to a final concentration of 12.5 mg/ml, aliquot to 50 ul each tube and store at -20C.
- PEG 400 from Sigma #P3265-1KG.
- 2% Tween 20, prepared in H2O.
- Prepare the working solution within 30 minutes of use.
ATTAC mouse model was first described by Scherer and colleagues. This strategy takes advantage of inducible ablation system to study function of particular cell types in vivo. ATTAC has been applied in several cell types inculding adipocytes, pancreatic beta cells and cardiac myocytes.
The dosage of dimerizer to obtain efficient ablation should be titrated for different animal models. Typically, concentration of o 0.2 - 1 ug/g body weight can be tried in a preliminary test.
Ref: Pajvani et al Nat Med, 2005. Wang et al Diabetes, 2008.
To obtain 0.2 ug/g body weight dosage, the working solution should be 0.05 ug/ul dimerizer. For a mouse of 30 g, inject 120 ul working solution traperitonealy.
For 1 ml working solution:
1.Add 100ul PEG-400 (100%) into a tube. Note: PEG is viscous. Make sure you transfer the exact amount.
2.Add 4 ul dimerizer to the tube with PEG and vortex.
3.Add 896ul 2% Tween and vortex. Keep on ice.
4.Inject within 30min.