Anti-Eck/EphA2 Antibody, clone D7

EPH and EPH-related receptors have been implicated in mediating developmental events, particularly in the nervous system. Receptors in the EPH subfamily typically have a single kinase domain and an extracellular region containing a Cys-rich domain and 2 fibronectin type III repeats. The ephrin receptors are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands.
Protein kinases are enzymes that transfer a phosphate group from a phosphate donor, generally the g phosphate of ATP, onto an acceptor amino acid in a substrate protein. By this basic mechanism, protein kinases mediate most of the signal transduction in eukaryotic cells, regulating cellular metabolism, transcription, cell cycle progression, cytoskeletal rearrangement and cell movement, apoptosis, and differentiation.

This antibody recognizes Epithelial Cell Kinase (Eck/EphA2), Mr 140 kDa

Native protein isolated by purification of phosphotyrosine-containing proteins. Clone D7.

Protein Kinase Assay:
A previous lot of this antibody has been reported to have been used in an immunoprecipitation autophosphorylation assay, using a Mn-PIPES reaction buffer (Romer, L., 1994).

Immunoprecipitation:
A previous lot of this antibody has been reported to immunoprecipitate Eck from 500 µg of a human breast epithelial cell line which had been lysed in TBS containing 1% Triton X-100. Use 1-4 µg per reaction.

Immunocytochemistry:
A previous lot of this antibody has been reported to immunostain Eck in human, mouse and rat epithelial cells fixed with 3.7% formaldehyde solution and permeabilized with 0.5% Triton X-100 in TBS.

Research Category
Neuroscience

Research Sub Category
Growth Cones & Axon Guidance

This Anti-Eck/EphA2 Antibody, clone D7 is validated for use in IC, IP, EA, WB for the detection of Eck/EphA2.

Routinely evaluated by immunoblot on RIPA lysates from human A431, foreskin fibroblasts, murine 3T3/A31 or rat L6 cells.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected Eck in RIPA lysates from human A431 and previously from foreskin fibroblasts, murine 3T3/A31 and rat L6 cells.

140 kDa

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1 in buffer containing 0.2 M Tris-glycine, pH 7.4, 0.15 M NaCl, with 0.05% sodium azide and 30% glycerol.

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. NOTE: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Control
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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