Anti-Rabbit HRP-DAB IHC detection kit

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 buffer for 3 x 3 minutes.

Incubate with HRP Goat anti-Rabbit IgG (H+L) (Ready-to-use, specific for IHC) for 30 minutes.

Wash in wash buffer 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 3 minutes, then rinse slides in water.

Clear and mount sections.

IHC shows positive staining in paraffin-embedded human kidney.
Anti-Vimentin antibody (S0B2254) was used at 1/500 dilution, followed by a HRP Goat anti-Rabbit IgG (H+L) (Ready-to-use, specific for IHC).
Counterstained with hematoxylin.
Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon.

Anti-EpCAM antibody (S0B2028) was used at 1/500 dilution, followed by a HRP Goat anti-Rabbit IgG (H+L) (Ready-to-use, specific for IHC).

Counterstained with hematoxylin.

Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.