Technical Data
Eck (EphA2, Mpk-5, Native Epithelial Cell Kinase, Eph receptor tyrosine kinase A2)
Suitable for use in Western Blot, Protein Kinase Assay, Immunoprecipitation and Immunocytochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 1-2ug/ml; Detected Eck in RIPA lysates from human A431 cells and previously from foreskin fibroblasts, murine 3T3/A31 and rat L6 cells. A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with E3360-01B. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system
Protein Kinase Assay: Reported to have been used in an IP autophosphorylation assay, using a Mn-PIPES reaction buffer.
Immunoprecipitation: Reported to immunoprecipitate Eck from 500ug of a human breast epithelial cell line which had been lysed in TBS containing 1% Triton X-100. Use 1-4ug per reaction.
Immunocytochemistry: 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.
Optimal dilutions to be determined by the researcher.

Recommended Secondary Antibody:
I1904-77B: IgG1 (HRP) Pab Gt x Mo

Positive Antigen Control:
A0001-50: Non-stimulated A431 lysate

Control cells:
CHO cell lysate, foreskin fibroblasts, 3T3/A31 cells, L6 cells.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
MabIgG10.T.34Affinity Purified
200ug4C (-20C Glycerol)Blue IceHumanMouse
Native protein isolated by purification of phosphotyrosine-containing proteins
Purified by Protein G affinity chromatography.
Supplied as a liquid in 0.2M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, before the addition of glycerol to 30%.
Recognizes human Epithelial Cell Kinase (Eck) at 140kD. Species Crossreactivity: mouse, rat, bovine and canine.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Romer, L., et al., Mol. Biol. Cell 5: 349-361 (1994). 2. Magal, E., et al., J. Neurosci. Res. 43: 735-744 (1996). 3. Pandey, A., et al., J. Biol. Chem. 270: 19,201-19,204 (1995). 4. Pandey, A., et al., Science 268: 567-569 (1995).