Description: CD68, Mouse, mAb FA-11

The monoclonal antibody FA-11 reacts with murine macrosialin (mouse CD68), a heavily glycosylated transmembrane protein of 87- 115 kDa, which is specifically expressed by tissue macrophages, Langerhans cells and at low levels by dendritic cells. Macrosialin belongs to the lysosomal-associated membrane protein (LAMP) family. In common with the LAMPs, macrosialin is a type I me.mbrane protein, containing a short and highly conserved cytoplasmic tail, followed by a transmembrane domain that precedes the intraluminal region. In macrophages, macrosialin is mainly expressed as a late endosomal protein and rapidly exchanges with a small subset of macrosialin present on the cell surface. Several reports have shown that macrosialin recognises oxidized low-density lipoproteins as well as the intercellular adhesive molecule (ICAM-L) raising the possibility of a receptor function of this protein. In human, macrosialin has been suggested to be a novel prognostic factor for classical Hodgkin’s lymphoma- since high expression of macrosialin and CD163 correlates with adverse outcome.The monoclonal antibody FA-11 detects surface macrosialin at low levels in resident mouse peritoneal macrophages which can be enhanced by thiolycollate stimulation. Macrosialin is predominantly located within the cell and can be detected by flow cytometry with the monoclonal antibody FA-11 when cell permeabilisation is used.

Specifications:

Catalog number Product type Quantity Formulation Application Application Notes Use Immunogen Isotype Species Alias Positive Control References Storage and stability Precautions Disease

HM1070

Monoclonal antibodies

100 µg

1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.

Flow cytometry, Frozen sections, Immuno fluorescence, Immuno precipitation, Western blot

FC: Antibody FA-11 stains both- intra and extracellular macrosialin. For intracellular staining- THP-1 cells were permeabilized with buffer containing 0.5% saponin. The THP-1 cells were treated an Fc receptor blocking solution to block nonspecific binding. As negative control an isotype matched control IgG was used. (Ref.3)W: A non-reduced sample treatment and SDS-Page was used. The band sizes iare 75-120kDa depending on glycosylation pattern of macrosialin. (Ref.5).IHC-F:- 5µm;M tissue sections were fixed in acetone for 10 minutes at room temperature. As- negative controls primary antibodies were omitted or replaced by rat mAbs to unrelated antigens (Ref.2).

For immunohistochemistry, flow cytometry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, in vitro dilutions have to be optimized in user’s experimental setting.

Purified glycoproteins from P815 cell line

Rat IgG2a

Mouse

CD68 antigen gp110

murine macrophage-like cell line RAW264.7 (Ref. 5)

1. Smith, M et al; Differential expression of murine macrophage surface glycoprotein antigens in intracellular membranes. J Cell Sci 1987, 87:1132. Rabinowitz, S et al; Macrosialin, a macrophage-restricted sialoprotein differentially glycosylated in response to inflammatory stimuli. J Exp Med 1991, 174: 8273. Ramprasad, M et al; Cell surface expression of mouse macrosialin and human CD68 and their role as macrophage receptors for oxidized low density lipoprotein. Proc Natl Acad Sci 1996, 93: 148334. Da Silva, R et al; Phagocytosis stimulates alternative glycolysation of macrosialin (mouse CD68), a macrophage-specific endosomal protein. Biochem J 1999, 338: 6875. De Beer, M et al; Lack of a direct role for macrosialin in oxidized LDL metabolism. J Lipid Res 2003, 44: 6746. Apostolopoulos, J et al; The cytoplasmic domain of tissue factor in macrophages augments cutaneous delayed-type hypersensitivity. J Leukoc Biol 2008, 83: 907. Rickard, A et al; Deletion of Mineralocorticoid Receptors From Macrophages Protects Against Deoxycorticosterone/Salt-Induced Cardiac Fibrosis and Increased Blood Pressure. Hypertension 2009, 54: 537

Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.

For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.

Tumor immunology

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