Biotinylated Goat Anti-Polyvalent (Ready-To-Use) (ab128977)

Procedure:

Step 1: Deparaffinize and rehydrate tissue section.

Step 2: (Optional) To reduce nonspecific background staining due to endogenous peroxidase (only when using peroxidase label), incubate slide in hydrogen peroxide for 10-15 minutes.

Step 3: Wash 2 times in buffer.

Step 4: If required, incubate tissue in digestive enzyme (or appropriate pretreatment).

Step 5: Wash 4 times in buffer.

Step 6: (Optional) Place slide in Ultra V Block and incubate for 5-10 minutes at room temperature to block nonspecific background staining. NOTE: Do not exceed 10 minutes or there may be a reduction in desired stain.

Step 7: Rinse (Optional).

Step 8: Apply primary antibody and incubate according to manufacturer's protocol.

Step 9: Wash 4 times in buffer.

Step 10: Apply Biotinylated Goat Anti-Polyvalent and incubate for 10-15 minutes at room temperature.

Step 11: Wash 4 times in buffer.

Step 12: Apply enzyme labelled streptavidin and incubate according to manufacturer’s protocol.

Step 13: Rinse 4 times in buffer.

Step 14: Apply appropriate chromogenic substrate and incubate until desired reaction is achieved.

Step 15: Counterstain and coverslip.

NOTE: This reagent is provided in bulk form for use with automated systems (including capillary gap methodologies) or in staining jars for repeated use. If the DIP method is employed, staining jar should be sealed and refrigerated between uses. Remove staining jars from the refrigerator 30 minutes prior to staining to allow reagents to come to room temperature. The specificity and sensitivity of antigen detection is dependent on the specific primary antibody used.

IHC Protocol Summary