A Highly Sensitive IHC Detection Protocol (HRP/DAB)

Polymer Enhanced Method

A. Jamsa from Carolinska University


GBI Polink Polymer Detection Systems






This protocol is good for detecting both mouse and rabbit primary antibodies on human tissues. It is very sensitive and stable.



Solution A (antibody linker)
Prepared properly, the solution will be stable for 18 months at 4 degrees. 
  • Rabbit anti-Mouse IgG (min x human serum protein) (Jackson ImmunoResearch Cat No. 315-005-045). Working concentration: 1-2ug/ml
  • Normal Goat Serum (Jackson ImmunoResearch Cat No. 005-000-121). Working concentration: 1%
  • Working Solution: Add concentrated rabbit anti-mouse IgG, normal goat serum, in antibody diluent (1% BSA, 0.1% sodium azide, in pH7.4 PBS) and make into working solution.
Solution B (polymer HRP anti-Rabbit)
You may buy this solution from many vendors: DAKO envision, Vector ImmPress etc.
After primary antibody incubation, apply Solution A on tissue slides and incubate 20 minutes under room temperature. Then wash in PBS wash buffer 3 minutes x 3. Apply Solution B and incubate for 30 minutes under RT, wash in PBS wash buffer 3 minutes x 3. Apply chromogen, and following other routine procedures.
Notes: This detection protocol is very sensitive for both mouse and rabbit primary antibodies. It is a good protocol for labs with routine IHC.