Tamoxifen
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Contents
Tamoxifen injection protocol
Method Summary
This method describes how to induce expression of cre-recombinase in tamoxifen-inducible transgenic Cre-mouse lines.
Materials
Tamoxifen
- 10mg/ml tamoxifen (final concentration)
Tamoxifen from Sigma, catalogue number: T5648.
Sesame or Peanut Oil
use either sesame or peanut oil
- Sesame Oil from Sigma, catalogue number: S3547
- Peanut Oil missing link
Method
- Weigh in 100mg of tamoxifen
- Resuspend the tamoxifen in 1ml of ethanol (100%) and mix; (this step will take some time; if necessary warm to 37ºC for 20-30 minutes)
- After tamoxifen is dissolved, add 9ml of sesame or peanut oil
- Mix on shaker
- Aliquot and store until use at -20ºC
Injection protocol
- Inject intra-peritoneal (IP) using either of the following dosages:
- 20mg/kg/day over 6 days,
- 30mg/kg/day over 5 days, or
- 80mg/kg/day over 2 (to 3) days.
Caution
Tamoxifen injection and cre expression may be linked to a transient cardiac hypertrophy reaction in mice. Consider using cre-positive wildtype mice as controls.
References:
- Cre-loxP DNA recombination is possible with only minimal unspecific transcriptional changes and without cardiomyopathy in Tg(alphaMHC-MerCreMer) mice.
- Dilated cardiomyopathy resulting from high-level myocardial expression of Cre recombinase.
- Avoidance of transient cardiomyopathy in cardiomyocyte-targeted tamoxifen-induced MerCreMer gene deletion models.