Cardiotoxin

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: [1].

BrdU

• BrdU

500 mg/kg at 1 mg/µl in normal saline

BrdU from Sigma, catalogue number: [2].

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Method

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

This procedure is adapted from the Yan Lab.