Technical datasheet
Description
ENA1 reacts with E-selectin CD62-E, previous designated the Endothelial Leucocyte Adhesion Molecule-1 (ELAM-1). The antibody reacts with human endothelial cells activated with TNF-alpha, IL-1 or endotoxin. The antibody was found to react also with cells transfected with the E-selectin gene. The antibody inhibits the adhesion of granulocytes both neutrophilic and eosinophilic.
Cross Reactivity
| Cross reactant |
Reactivity |
| Baboon |
Yes |
| Chimpanzee |
Yes |
| Cynomolgus monkey |
Yes |
| Rhesus monkey |
Yes |
Formulation
0.5 mg of 0.2 μm filtered antibody solution in PBS (exact concentration is indicated on the label), containing 0.02% sodium azide.
Species
Mouse IgG1
Application
The antibody is useful for staining of E-selectin expressing endothelial cells. It permits staining of in vitro cultured cells as well as frozen sections. The antibody can also be used for staining of chimpanzee, rhesus monkey, cynomolgous monkey and baboon endothelial cells expressing E-selectin. Furthermore the antibody can be used for immuno precipitation and immuno assays. For adhesion inhibition studies we advise to use the F(ab')2 ENA2 monoclonal antibody preparation (Cat. nr. HM4003), which is most efficient for this purpose and prevents adhesion of leucocytes via Fc receptors.
Use
For immunohistology 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 1:10. In vitro cultured cells can be fixed with 1% paraformaldehyde and kept in PBS plus azide before staining. Tissue sections are advised to be fixed for 10 min in pure acetone and followed by incubation for 10 min in chloroform. Incubation with a pretested dilution of the antibody is advised to be followed by a biotin conjugated anti-murine Ig and a further incubation with an enzyme (alkaline phosphatase) conjugated streptavidin. For selection of the most useful dilution in a given situation a test staining with cells or tissue known to express the antigen should be performed. To this end either cultured endothelial cells or a small fresh skin biopsy can be incubated for 4 hours with TNF-alpha (1 ng/ml), IL-1 (100 U/ml) or LPS (1 μg/ml) in tissue culture medium at 37˚C. As negative control it is advised to use a control murine IgG1 antibody.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
- Leeuwenberg, JFM et al; Induction of an activation antigen on human endothelial cells in vitro. Eur J Immunol 1989, 19: 715
- Leeuwenberg, JFM et al; Adhesion of polymorphonuclear cells to human endothelial cells. Adhesionmolecule dependent, and Fc-receptor-mediated adhesion-molecule-independent mechanisms. Clin and Exp Immunol 1990, 81: 496
- Leeuwenberg, JFM et al; Functional polymorphism of IgG FcRII (CD32) on human neutrophils. Immunology 1990, 71: 301
- Leeuwenberg, JFM et al; Interferon-gamma regulates the expression of an adhesion molecule ELAM-1 and IL-6 production by human endothelial cells in vitro. J Immunology 1990, 145: 2110
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 with the use of or derivation of this product.
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