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Anti-Rabbit and Mouse HRP-DAB IHC detection kit (2-step) utilized the newest biotin-free polymerization technology to prepare super sensitive polymeric peroxidase linked conjugates. It utilizes a novel controlled and compact enzyme polymerization technology to achieve higher sensitivity without background. It provides on step detection protocol, superior sensitivity and specificity, short incubation time and faster turnaround.
12 months from date of receipt / reconstitution, 2 to 8 °C as supplied
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Immunohistochemistry protocol:
Cut and mount sections on slides coated with a suitable tissue adhesive.
De-paraffinize sections in xylene or xylene substitutes.
Re-hydrate through graded alcohols.
Wash slides in distilled H2O.
Perform antigen retrieval as required.
Wash slides in distilled H2O.
Quench endogenous peroxidase using Hydrogen Peroxide Block for 5-10 minutes.
Wash in wash buffer for 3 x 3 minutes.
Incubate with optimally diluted primary antibody.
Wash in wash solution for 3 x 3 minutes.
Incubate with Enhancer (A) for 10-15 minutes (RT). Wash in wash solution for 3 x 3 minutes.
Incubate with ready to use Goat anti-Rabbit & Mouse HRP Conjugate (B) for 15-20 minutes. Wash in wash solution for 3 x 3 minutes.
Add DAB working solution (DAB substrate diluted with DAB dilution buffer) until color development finish.
Rinse slides in water. Add 100μl Haematoxylin and incubate for 1-3 minutes, then rinse slides in water.
Clear and mount sections.
IHC shows positive staining in paraffin-embedded human tonsil. Anti-CD14 antibody (S0B2221) was used at 1/500 dilution, followed by anti-Rabbit and Mouse HRP-DAB IHC detection kit (2-step) (S0C2011). Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.