05-480
Anti-Eck/EphA2 Antibody, clone D7
clone D7, Upstate®, from mouse
Synonym(s):
EPH receptor A2, Epithelial cell kinase, Tyrosine-protein kinase receptor ECK, ephrin receptor EphA2, epithelial cell receptor protein tyrosine kinase, protein tyrosine kinase, receptor protein tyrosine kinase regulated by p53 and E2F-1
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About This Item
UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41
Recommended Products
biological source
mouse
Quality Level
antibody form
purified immunoglobulin
antibody product type
primary antibodies
clone
D7, monoclonal
species reactivity
mouse, canine, human, rat, bovine
manufacturer/tradename
Upstate®
technique(s)
activity assay: suitable
immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable
isotype
IgG1
NCBI accession no.
UniProt accession no.
shipped in
dry ice
target post-translational modification
unmodified
Gene Information
bovine ... Epha2(512798)
human ... EPHA2(1969)
General description
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.
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.
Specificity
This antibody recognizes Epithelial Cell Kinase (Eck/EphA2), Mr 140 kDa
Immunogen
Native protein isolated by purification of phosphotyrosine-containing proteins. Clone D7.
Application
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.
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
Neuroscience
Research Sub Category
Growth Cones & Axon Guidance
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.
Quality
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.
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.
Target description
140 kDa
Physical form
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.
Storage and Stability
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.
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.
Analysis Note
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.
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.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Find documentation for the products that you have recently purchased in the Document Library.
Tyrosine kinase activity, cytoskeletal organization, and motility in human vascular endothelial cells.
Romer, L H, et al.
Molecular Biology of the Cell, 5, 349-361 (1994)
Antibody targeting of the EphA2 tyrosine kinase inhibits malignant cell behavior.
Carles-Kinch, Kelly, et al.
Cancer Research, 62, 2840-2847 (2002)
Identification of tyrosine phosphorylated adhesion proteins in human cancer cells.
Kinch, M S, et al.
Hybridoma, 17, 227-235 (1998)
Christine Gundry et al.
Nature communications, 8, 14646-14646 (2017-03-16)
The Rab GTPase effector, Rab-coupling protein (RCP) is known to promote invasive behaviour in vitro by controlling integrin and receptor tyrosine kinase (RTK) trafficking, but how RCP influences metastasis in vivo is unclear. Here we identify an RTK of the
Predictive value of the EphA2 receptor tyrosine kinase in lung cancer recurrence and survival.
Michael S Kinch, Mary-Beth Moore, David H Harpole
Clinical cancer research : an official journal of the American Association for Cancer Research null
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