Technical datasheet
Description
Monoclonal antibody FA6-152 recognizes human CD36 (88-kDa), a cell surface class B scavenger receptor, also known as thrombospondin receptor CD36 is a heavily N-glycosylated transmembrane protein of ~88 kDa with two short intracellular domains and a large extracellular domain. The protein is sensitive for neuroaminidase, resulting in a shift from 88 to 85 kDa. CD36 is expressed on platelets, mature monocytes and macrophages, microvascular endothelial cells, mammary endothelial cells, during stages of erythroid cell development and on some macrophage derived dendritic cells. The antibody recognizes adult and fetal monocytes, platelets and reticulocytes, but doesn’t stain lymphocytes and granulocytes. Reactivity has also been found in small intestine, kidney, liver and thyroid. CD36 expression is primarily controlled by the transcription heterodimer PPARg-RXR (peroxisome proliferator-activated receptor-g-retinoid-X-receptor). CD36 is preferentially found within lipid rafts, which facilitates its association with receptors, signaling and adaptor molecules. It is a receptor and transporter of oxidized lipids and long chain fatty acids. CD36 has been implicated in many biological processes including angiogenesis, phagocytosis, inflammation, and lipid and glucose metabolism. Several in vivo models support the role of the thrombospondin / CD36 system in angiogenesis and tumor growth. An important role for CD36 has been found in Malaria as major receptor for P. falciparum-infected red blood cells. CD36 is associated with Src-family kinases and with the integrins α3β1 and α6β1. Recently, CD36 has been identified as a protein that is required for toll like receptor (TLR2) recognition of di-acylated bacterial lipopeptides and lipoteichoic acid4. Furthermore, CD36 has been shown to function as phagocytic receptor for apoptotic cells. Many different ligands have been reported to interact with CD36, suggesting that CD36 could recognize a structure-based domain rather than specific contact residues. Monoclonal antibody FA6-152 blocks the biological activity of CD36 by blocking collagen/thrombospondin binding. The antibody agglutinates fetal but not adult erythrocytes.
Immunogen
20-Weeks-old fetal erythrocytes
Formulation
0.5 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin
Species
Mouse IgG1
Application
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F4,5
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FC1
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FS2
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IA3
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IF1,4
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IP2
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P
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W
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Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application flow cytometry has been tested by Hycult Biotech.
Application notes
F: Tissue embedded in tissue-tek (for instance aortic tissue) followed by freezing in liquid nitrogen; 7-8 µm sections; air-dried; aceton-fixed; 10 % NGS as block (Ref 4).
FC: Antibody FA6-152 stains the extracellular domain of CD36. Unfixed cells; 2µg per 100.000 cells. Positive on granulocytes (Ref 1).
FS: Platelet aggregation and secretion was induced by > 1µg/ml antibody (Ref 2).
IA: 10 µg/ml antibody as coat diluted in Tris-buffered saline ; 100 µl/well; o/n at RT (Ref 3).
IF: unfixed cells were incubated for 30 minutes at 4 °C followed by a secondary FITC polyclonal antibody; one-minute methanol fixation before analysis (Ref 1).
IP: 88 kDa sialoglycoprotein in platelets; 85 and 88 kDa in HEL cells. 10 µg antibody/200 µg protein.
Use
For immunohistology and flow cytometry, 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.
Aliases
GPIV, FAT, platelet glycoprotein 4, thrombospondin receptor
Positive control
HELA cells
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.
References
1. Edelman P et al; A monoclonal antibody against an erythrocyte ontogenic antigen identifies fetal and adult erythroid progenitors. Blood 1986, 67: 56.
2. Kieffer N et al; Development regulated expression of a 78 kDa erythroblast membrane glycoprotein immunologically related to the platelet thrombospondin receptor. Biochem J 1989, 262: 835
3. Thibert V et al; Quantitation of platelet glycoprotein IV (CD36) in healthy subjects and in patients with essential thrombocythemia using an immunocapture assay. Thromb Haemost 1992, 68: 600
4. Nakata, A et al; CD36, a novel receptor for oxidized low-density lipoproteins, is highly expressed on lidip-laden macrophages in human atherosclerotic aorta. Arterioscler Thromb Vasc Biol 1999, 19: 1333
5. Eharta, S et al; Pathophysiological role of oxidezed low-density lipoprotein in plaque instability in coronary artery diseases. J Diab Complic 2002, 16: 60
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.
References
1. Edelman P et al; A monoclonal antibody against an erythrocyte ontogenic antigen identifies fetal and adult erythroid progenitors. Blood 1986, 67: 56.
2. Kieffer N et al; Development regulated expression of a 78 kDa erythroblast membrane glycoprotein immunologically related to the platelet thrombospondin receptor. Biochem J 1989, 262: 835
3. Thibert V et al; Quantitation of platelet glycoprotein IV (CD36) in healthy subjects and in patients with essential thrombocythemia using an immunocapture assay. Thromb Haemost 1992, 68: 600
4. Nakata, A et al; CD36, a novel receptor for oxidized low-density lipoproteins, is highly expressed on lidip-laden macrophages in human atherosclerotic aorta. Arterioscler Thromb Vasc Biol 1999, 19: 1333
5. Eharta, S et al; Pathophysiological role of oxidezed low-density lipoprotein in plaque instability in coronary artery diseases. J Diab Complic 2002, 16: 60