|The ESAM esam (Catalog #MBS601874) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The ESAM (Endothelial Cell Adhesion Molecule) reacts with Human and may cross-react with other species as described in the data sheet. MyBioSource\'s ESAM can be used in a range of immunoassay formats including, but not limited to, ELISA (EL/EIA), Western Blot (WB), Immunocytochemistry (ICC), Flow Cytometry (FC/FACS).
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. Researchers should empirically determine the suitability of the ESAM esam for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process.
The ESAM esam product has the following accession number(s) (GI #148664205) (NCBI Accession #NP_620411.2) (Uniprot Accession #Q96AP7). Researchers may be interested in using Bioinformatics databases such as those available at The National Center for Biotechnology Information (NCBI) website for more information about accession numbers and the proteins they represent. Even researchers unfamiliar with bioinformatics databases will find the NCBI databases to be quite user friendly and useful.
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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.
Immunogen: NS0-derived rhESAM.