View source for Cardiotoxin

== Cardiotoxin injection protocol ==

=== Method Summary ===
This method describes how to induce regeneration of skeletal muscle by intramuscular injection of cardiotoxin.

=== Materials ===

==== Cardiotoxin====
* Cardiotoxin is purchased from CalBiochem (1mg)
Stock solution: 10 µM dissolve in 13.9 ml of PBS and filter through a 2 µ filter (store at 4ºC up to 6 months). 

Cardiotoxin from CalBiochem, catalogue number 217504: [https://www.emdmillipore.com/INTL/en/product/cardiotoxin-naja-nigricollis,EMD_BIO-217504].

==== BrdU====
* BrdU
500 mg/kg at 1 mg/µl in normal saline
 
BrdU from Sigma, catalogue number: [http://www.sigmaaldrich.com/catalog/product/sigma/b5002?lang=en&region=US].



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=== Method ===

# Restrain the hindlimb and shave to remove hair.
# Wet hindlimb with 70% ethanol.
# Inject 0.1 ml of 10 µM cardiotoxin intramuscularly into the tibialis anterior muscle(s).
# Six hours before harvesting, inject BrdU (Sigma B5002) (500 mg/kg at 1mg/µl in normal saline) i.p.
# Harvest muscle at day 1, 2, 3, 5, 10, and 14.


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This procedure is adapted from the Yan Lab.