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Catalog # HM4003

E-selectin, Human, mAb ENA2

125.00390.00

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ENA2 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.

Application
Frozen sections, Functional studies, Immuno assays, Immuno fluorescence

Application Notes
FS: the antibody can be used for adhesion inhibition studies for which the antibody is most efficient and prevents adhesion of leucocytes via Fc receptors. (Ref.3) IF: 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.

Use
For immunohistochemistry, 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
Fresh skin biopsy 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.

Product type
Monoclonal antibodies
Amount
100 µg, 20 µg
Formulation
0.2 ml (100 μg/ml) 0.2 μm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.
Isotype
Mouse IgG1
Species
Human
Cross reactivity
Baboon – Yes, Chimpanzee – Yes, Cynomolgus monkey – Yes, Rhesus monkey – Yes
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for one year.
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.
Disease
Cardiology and metabolism, Neurological disorders
Protocol CoA-TDS HM4000-6000
HM4003
81 kb
Safety Data Sheet

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