Difference between revisions of "Tamoxifen"

From SDMRC
Jump to: navigation, search
 
(3 intermediate revisions by the same user not shown)
Line 8: Line 8:
  
 
==== Tamoxifen ====
 
==== Tamoxifen ====
* 100mg/10ml tamoxifen (final concentration)
+
* 10mg/ml tamoxifen (final concentration)
  
 
Tamoxifen from Sigma, catalogue number: [http://www.sigmaaldrich.com/catalog/product/sigma/t5648?lang=en&region=US T5648].
 
Tamoxifen from Sigma, catalogue number: [http://www.sigmaaldrich.com/catalog/product/sigma/t5648?lang=en&region=US T5648].
Line 34: Line 34:
 
----
 
----
  
=== CAUTION ===
+
=== Caution ===
tamoxifen injection and cre expression may be linked to a transient hypertrophy reaction in mice.<br />
+
Tamoxifen injection and cre expression may be linked to a transient cardiac hypertrophy reaction in mice. Consider using cre-positive wildtype mice as controls.<br />
References:
+
'''References:'''
 
# Cre-loxP DNA recombination is possible with only minimal unspecific transcriptional changes and without cardiomyopathy in [http://www.ncbi.nlm.nih.gov/pubmed/20802136 Tg(alphaMHC-MerCreMer)] mice.
 
# Cre-loxP DNA recombination is possible with only minimal unspecific transcriptional changes and without cardiomyopathy in [http://www.ncbi.nlm.nih.gov/pubmed/20802136 Tg(alphaMHC-MerCreMer)] mice.
 
# Dilated cardiomyopathy resulting from high-level myocardial expression of [http://www.ncbi.nlm.nih.gov/pubmed/16762803?access_num=16762803&link_type=MED&dopt=Abstract Cre recombinase].
 
# Dilated cardiomyopathy resulting from high-level myocardial expression of [http://www.ncbi.nlm.nih.gov/pubmed/16762803?access_num=16762803&link_type=MED&dopt=Abstract Cre recombinase].
 +
# Avoidance of transient cardiomyopathy in cardiomyocyte-targeted [http://www.ncbi.nlm.nih.gov/pubmed/19520971 tamoxifen-induced MerCreMer gene deletion models.]

Latest revision as of 00:11, 11 November 2014

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

  1. Weigh in 100mg of tamoxifen
  2. 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)
  3. After tamoxifen is dissolved, add 9ml of sesame or peanut oil
  4. Mix on shaker
  5. Aliquot and store until use at -20ºC

Injection protocol

  • Inject intra-peritoneal (IP) using either of the following dosages:
    1. 20mg/kg/day over 6 days,
    2. 30mg/kg/day over 5 days, or
    3. 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:

  1. Cre-loxP DNA recombination is possible with only minimal unspecific transcriptional changes and without cardiomyopathy in Tg(alphaMHC-MerCreMer) mice.
  2. Dilated cardiomyopathy resulting from high-level myocardial expression of Cre recombinase.
  3. Avoidance of transient cardiomyopathy in cardiomyocyte-targeted tamoxifen-induced MerCreMer gene deletion models.