Usage
Western Blot: This antibody can be used at 1-2 ug/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 25 ug/ml and add 10 ul of the diluted solution to 1-2.5 x 10^5 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 10 ug/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 0.5-1.0 ug/ml with the appropriate secondary reagents to detect human ESAM. The detection limit for rhESAM is approximately 1 ng/well.
Reconstitution
Reconstitute in deionized water
Restrictions
For research use only. Intended for use by laboratory professionals.
Guarantee
This antibody carries the LSBio 100% Guarantee.
LSBio Guarantee