| 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. |
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| Catalog # | E3486-50 |
| Hybridoma | Produced 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). |
| Applications | Western 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 Stability | Lyophilized 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. |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG2b |
| Clone No | 7H234 |
| Host | Mouse |
| Source | Human |
| Concentration | As reported |
| Form | Supplied 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. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | NS0-derived rhESAM |
| Specificity | Selected for its ability to detect human ESAM in direct ELISAs and Western Blots. In these applications, this antibody shows no crossreactivity with rmESAM. |
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| Important Note | This 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. |
