DEN Protocol

DEN‐induced HCC model in male mice

Zhong Xu, January 2016

Ordering information

Items Catalog # MFR
DEN N0756-10 ML Sigma
HFD 14726603 Fisher Scientific Company, LLC


  1. prepared 5 ug/ul DEN working solution (26.4 μl DEN +5 ml sterilized PBS)
    [26.4 ul DEN =25 .08 mg DEN (density=0.95, Mass= Volume X Density)]
  2. The mice were injected intraperitoneally with DEN (25 mg/kg) at 14‐day age.
  3. Wean and keep male mice at 21‐day age.
  4. Identify gene type (for instance ACR+, ACR‐) by genotyping.
  5. After 4 weeks, mice at 6 weeks age were fed with HFD.
  6. At 33 weeks, mice were scarified for H&E, IHC and Western blot, etc.


Mice at 6 weeks age were separated into two groups. One group was fed with standard diet, another was fed with HFD. Then mice were scarified at 33 weeks.

HFD group


SD group