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Catalog # HM1070

CD68, Mouse, mAb FA-11

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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.

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

Application Notes
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:- 5 µm;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).

Use
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.

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

Product type
Monoclonal antibodies
Quantity
20 µg, 100 µg
Formulation
0.2 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Immunogen
Purified glycoproteins from P815 cell line
Isotype
Rat IgG2a
Species
Mouse
Alias
CD68 antigen gp110
Storage and stability
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.
Precautions
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.
Disease
Tumor immunology

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