Mounting Medium for IHC - Aqueous

Mounting Medium ab64230 is an aqueous mounting medium designed for use with chromogenic tissue and cell staining. It can be used with chromogens such as Fast-Red or AEC that require an aqueous mounting medium, or those that do not, such as DAB. When it is dry, it generates an even surface and can be used without a glass coverslip.

Mounting media products

For fluorescent cell and tissue staining, Abcam recommends aqueous, anti-fade Fluorescence Mounting Medium ab104135, Mounting Medium with DAPI ab104139, and Mounting Medium with PI ab104129. For thick sections or tissues containing lots of fat, we recommend Glycerol Mounting Medium with DAPI ab188804.

For chromogenic immunohistochemistry, such as with DAB, AEC, or Fast Red, we recommend this product, Mounting Medium ab64230, or organic Limonene Mounting Medium ab104141.

IHC protocol suitable for use with Mounting Medium ab64230:

1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.

2. Use appropriate antigen retrieval buffer or enzyme (primary antibody dependent) to treat sections. Wash 3 times in buffer.

3. Add enough hydrogen peroxide blocking solution to cover the sections. Incubate for 10 minutes. Wash 2 times in buffer. If necessary, use avidin biotin blocking.

4. Apply protein block (or normal serum from same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.

5. Apply primary antibody in antibody diluent and incubate.

6. Wash 4 times in buffer. Incubate slide with biotinylated secondary antibody (or HRP polymer secondary antibody and skip step 7). Wash 4 times in buffer.

7. Apply streptavidin-HRP and incubate for 10 minutes at room temperature.

8. Rinse 4 times in buffer. Place slide in DAB substrate or AEC Substrate and incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.

9. Add enough drops of hematoxylin to cover the section. Incubate for 1 minute.

10. Rinse 7-8 times in tap water. Add Mounting Medium ab64230 to cover the section.

Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.