Application: TMB-IHC chromogen is very sensitive. It is recommended that the peroxidase should be diluted (approx. 16X) compared to DAB used as chromogen, e.g. if peroxidase is diluted 1:100 for DAB, one should use approx. 1:1600 dilution of peroxidase when TMB is used.
- After the peroxidase reaction, wash slides thoroughly with PBS or Tris-HCl saline containing 0.1 % Tween 20. DO NOT ADD THIMEROSAL, azide or phenolic compounds as preservative in any buffers.
- Add TMB-IHC solution and incubate for 5-15 minutes at room temp.
- Stop reaction by washing thoroughly with distilled or deionized water.
- Counterstain with Neutral Red solution 30-60 seconds.
- Rinse slides with distilled water, dehydrate with gradient of alcohols, xylene or any other clearing agent.
- Mount slides with organic mounting medium or our Organo/Limonene mount (AR-6504).
- Brilliant blue color is observed with TMB.