The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesDot: 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.
FunctionReceptor for members of the ephrin-B family. Binds to ephrin-B2. May have a role in events mediating differentiation and development.
Tissue specificityAbundantly expressed in placenta and in a range of primary tissues and malignant cell lines. Expressed in fetal, but not adult, brain, and in primitive and myeloid, but not lymphoid, hematopoietic cells.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. Ephrin receptor subfamily. Contains 2 fibronectin type-III domains. Contains 1 protein kinase domain. Contains 1 SAM (sterile alpha motif) domain.