“Collagenase Media“

1)      HG-DMEM

2)      P / S ( anti-biotic) ß (5ml/500mL)

 

Culture media

1)      HG-DMEM

2)      P / S (anti-biotic) (5ml/500mL)

3)      10% Serum (FBS)(50 mL in 1 bottle)

4)      HEPES – 1.19grams

5)      NaOH – add to reach 7.4 pH

 

To make the Collagenase solution

  1. Dilute Sigma Collagenase type XI to 20 U/ml in ‘Collagenase Media’
    1. Eg.- 100 mg at 2.2 U/mg = add 11 ml to the bottle
  2. Alloquot 1 ml each to 15 ml Falcon tubes and freeze at -20
  3. Before use, add 9 ml of ‘Collagenase Media’ to make 10 ml (at 2 U/ml)
  4. In general for mice, inject 5 ml and for rats, inject 10 ml