Eck (EphA2, Mpk-5, Native Epithelial Cell Kinase, Eph receptor tyrosine kinase A2)

Applications: \nSuitable for use in Western Blot, Protein Kinase Assay, Immunoprecipitation and Immunocytochemistry. Other applications not tested.\n\nRecommended Dilution:\nWestern 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\nProtein Kinase Assay: Reported to have been used in an IP autophosphorylation assay, using a Mn-PIPES reaction buffer.\nImmunoprecipitation: 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.\nImmunocytochemistry: 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.\nOptimal dilutions to be determined by the researcher.\n\nRecommended Secondary Antibody:\nI1904-77B: IgG1 (HRP) Pab Gt x Mo\n\nPositive Antigen Control: \nA0001-50: Non-stimulated A431 lysate\n\nControl cells:\nCHO cell lysate, foreskin fibroblasts, 3T3/A31 cells, L6 cells.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. 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.

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SPECIFICATIONS

Catalog Number

E3360-01B

Size

200ug

Applications

ICC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Rat, Bov, Can

Form

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%.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG1

References

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).

Additional Info

Recognizes human Epithelial Cell Kinase (Eck) at 140kD. \nSpecies Crossreactivity: mouse, rat, bovine and canine.