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You are here:Home » Antibodies » Abs to Carbohydrates, Glycoproteins » Anti -ESAM (Endothelial Cell Adhesion Molecule)

Anti -ESAM (Endothelial Cell Adhesion Molecule)

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Specifications

Clone Host Grade Applications
Monoclonal Mouse Affinity Purified E B IC FC
Endothelial cell adhesion molecule (ESAM) is a 55kD type I transmembrane glycoprotein of the JAM family of immunoglobulin superfamily molecules. The 390 aa human ESAM contains a 216 aa extracellular domain (ECD) with a V-type and a C2-type Ig domain. The ECD of human and mouse ESAM share 69% aa identity. ESAM is expressed on endothelial cells, activated platelets and megakaryocytes. ESAM mediates endothelial cell homophilic adhesion and supports neutrophil extravasation through tight junctions.
Catalog #E3486-50
HybridomaProduced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, NS0-derived, recombinant human ESAM (rhESAM; aa30-247; Accession # NP_620411).
ApplicationsWestern Blot: This antibody can be used at 1-2ug/ml with the appropriate secondary reagents to detect human ESAM. Using a colorimetric detection system, the detection limit for rhESAM is approximately 25 ng/lane under non-reducing and reducing conditions. Chemiluminescent detection will increase sensitivity by 5 to 50 fold.
Flow Cytometry: This antibody was validated for flow cytometry using HUVECs. Dilute this antibody to 25ug/ml and add 10 L of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200 L. The binding of unlabeled monoclonal antibodies may be visualized by adding a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome. HUVECs were stained with anti-ESAM or isotype control antibody, followed by PE-conjugated anti-mouse antibody.
Immunocytochemistry: This antibody was used at a concentration of 10ug/ml to detect ESAM in HUVECs. Cells were fixed with PBS containing 4% paraformaldehyde and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA. After blocking, cells were incubated with diluted primary antibody followed by NL557-coupled anti-mouse IgG in the dark. Between each step, cells were washed with PBS containing BSA.
Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human ESAM. The detection limit for rhESAM is approximately 1 ng/well. Optimal dilutions should be determined by each laboratory for each application.
Storage and StabilityLyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Clone TypeMonoclonal
IsotypeIgG2b
Clone No7H234
HostMouse
SourceHuman
ConcentrationAs reported
FormSupplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.2ml is used, the concentration will be 0.5mg/ml.
PurityPurified by Protein G affinity chromatography.
ImmunogenNS0-derived rhESAM
SpecificitySelected for its ability to detect human ESAM in direct ELISAs and Western Blots. In these applications, this antibody shows no crossreactivity with rmESAM.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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