A Highly Sensitive IHC Detection Protocol
Polymer Enhanced Method
A. Jamsa from Carolinska University
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
- Rabbit anti-Mouse IgG (min x human serum protein)
(Jackson ImmunoResearch Cat No. 315-005-045). Working
- 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
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.
detection protocol is very sensitive for both mouse and rabbit
primary antibodies. It is a good protocol for labs with routine