Tetramethylbenizidine IHC Peroxidase

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. 1. 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. 2. Add TMB-IHC solution and incubate for 5-15 minutes at room temp. 3. Stop reaction by washing thoroughly with distilled or deionized water. 4. Counterstain with Neutral Red solution 30-60 seconds. 5. Rinse slides with distilled water, dehydrate with gradient of alcohols, xylene or any other clearing agent. 6. Mount slides with organic mounting medium or our Organo/Limonene mount. 7. Brilliant blue color is observed with TMB.