Macrophage/HAM-56

Macrophages comprise many forms of mononuclear phagocytes found in tissues that derive from hematopoietic stem cells in the bone marrow. Among the functions of macrophages are nonspecific phagocytosis and pinocytosis, killing of ingested microorganisms, and digestion and presentation of antigens to T and B-lymphocytes. Macrophages work to secrete a large number of diverse products such as lysozyme and collagenases, several complement components and coagulation factors, some prostaglandins and leukotrienes, and many regulatory molecules (Interferon, Interleukin 1).

Macrophage HAM-56 reacts with tingible macrophages (found in the germinal centers of lymph nodes), interdigitating macrophages of lymph nodes and tissue macrophages, (e.g., Kupffer cells of the liver and alveolar macrophages of the lung). This antibody also stains a subpopulation of endothelial cells, most prominently those of the capillaries and smaller blood vessels. HAM-56 reacts with monocytes, but is unreactive with B and T-lymphocytes.

Macrophage HAM-56 is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

1. Gown AM, et al. Am J Pathol. 125:191-207
2. Alpers CE, et al. Am J Pathol. 92:662-665

1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
5. Any of three heating methods may be used:
   • Electric Pressure Cooker
     Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
   • Water Bath Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
   • Conventional Steamer Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.