This anti-macrophage monoclonal antibody, AM-3K, was produced by using human alveolar macrophages as immunogen. AM-3K reacts intensely with most of macrophages in lymphoreticular organs and in many other organs and tissues. AM-3K also reacts with the macrophages in many pathological conditions. However, this antibody does not react with dendritic cell population, such as epidermal Langerhans cells, interdigitating cells in the paracortex of lymph nodes, nor follicular dendritic cells. Lymphocytes, granulocytes and freshly isolated blood monocytes are also negative. Reaction products for AM-3K were found on the cytoplasmic membrane of macrophages by immunoelectron microscopy.
In both cryostat sections and formalin-fixed paraffin sections, this antibody recognizes the antigen presenting on the cell surface membrane of tissue macrophages, but not monocytes or dendritic cells.
The molecular weights of the antigen recognized by AM-3K are 120 and 70 kDa.
Human liver (paraffin section)
Kupffer cells are positivery stained
Human lymph node (paraffin section)
Macrophages in marginal sinus are positivery stained.
|Takeya M., Second Department of Pathology, Kumamoto
University School of Medicine, Kumamoto, Japan
|Thymus||Macrophages in cortex
Macrophages in medulla
|Spleen||Red pulp macrophages
White pulp marginal zone macrophages
|IDCs in PALS|
|Lymph nodes||TB macrophages in follicles
|IDCs in paracortical areas|
|Skin||Dermal macrophages||Langerhans cells|
PALS=periarteriolar lymphatic sheath; TB=tingible body; IDCs=interdigitating cells
Positive: Human, Monkey, Horse, Bovine, Pig, Goat, Dog, Cat, Rabbit, Guinea pig
|Package Size||50μg (200μL / vial)|
|Format||Mouse monoclonal antibody 0.25mg/mL|
|Buffer||Block Ace as a stabilizer, containing 0.1% Proclin as a bacteriostat|
|Storage||Store below 20°C
Once thawed, store at 4°C. Repeated freeze-thaw cycles should be avoided
|Purification method||The spleen cells from BALB/c mouse, immunized with human alveolar macrophages, were fused to myeloma NS-1 cells. The screening of the hybridoma cells was performed on cryostat sections of human lung. The cell line (AM-3K) with positive reaction was grown in ascitic fluid of BALB/c mouse, from which the antibody was purified by Protein G affinity chromatography.|
|Working dilution for immunohistochemistry: 10μg/mL, on frozen sections and paraffin sections.
Antigen retrieval (microwave 10min, 0.01M citrate buffer, pH2.0) recommended.
1. Zeng L., Takeya M., and Takahashi K. (1996) AM-3K, A novel monoclonal antibody specific for tissue macrophages and its applications to pathological investigation. Journal of Pathology 178 : 207-214
2. Yamate J., Yoshida H., Tsukamoto Y., Ide M., Kuwamura M., Ohashi F., Miyamoto T., Kotani T., Sakuma S., Takeya M. (2000) Distribution of cells immunopositive for AM-3K, a novel monoclonal antibody recognizing human macrophages, in normal and diseased tissues of dogs, cats, horses, cattle, pigs and rabbits. Vet Pathol 37(2): 168-176
3. Zeng L., Takeya M., Ling X., Nagasaki A., Takahashi K. (1996) Interspecies reactivities of anti-human macropharge monoclonal antibodies to various animal species. J Histochem Cytochem 44(8): 845-853
4. Frangogiannis NG., Burns AR., Micheal LH., Entman ML. (1999) Histochemical and morphological characteristics of canine cardiac mast cells. Histochem J 31(4):221-229
Anti Macrophage Surface Antigen