Anti-Eph receptor B4 antibody (ab10618)

Application notes

Dot: Use at a concentration of 0.1 - 0.2 µg/ml. The detection limit for recombinant mouse EphB4 is approximately 5 ng/lane under non-reducing and reducing conditions.
ELISA: Use at a concentration of 0.5 - 1.0 µg/ml, detection limit 1.5 ng/well.
Inhib: Use at a concentration of 0.5 - 2 µg/ml. For neutralization of receptor-ligand interaction, approximately 0.5-2 µg/ml of the antibody will block 50% of the binding of recombinant mouse ephrin-B2/ (5 ng/ml) to immobilized recombinant mouse EphB4/Fc chimera (100 µl of a 2 µg/ml solution coated in each well).

The recombinant mouse EphB4 used for the testing of this antibody consists of the extracellular domain of mouse EphB4 (amino acids 1-539) fused by means of a polypeptide linker to the Fc portion of human IgG1 that is histidine-tagged at the C-terminus. N-terminal sequencing indicates that recombinant mouse EphB4/Fc has Leu 16 at the amino terminus. The calculated molecular mass of the reduced protein is approximately 85.2 kDa, but as a result of glycosylation, recombinant EphB4/Fc migrates as an approximately 110 kDa protein under reducing conditions in SDS-PAGE. EphB4 binds to ephrin-B1 and ephrin-B2. Human and mouse EphB4 extracellular domains share approximately 88% homology.

Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.