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Anti-Eck/EphA2, clone D7
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R, M, B, Ca | IC, IP, EA, WB | Mouse | Purified | Monoclonal Antibody |
Description:
Anti-Eck/EphA2, clone D7
Trade Name:
Upstate (Millipore)
Specificity:
This antibody recognizes Epithelial Cell Kinase (Eck/EphA2), Mr 140 kDa
Molecular Weight:
140 kDa
Immunogen:
Native protein isolated by purification of phosphotyrosine-containing proteins. Clone D7.
Clone:
D7
Isotype:
IgG1
Background Information:
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.
Species Reactivity:
- Human
- Rat
- Mouse
- Bovine
- Canine
Species Reactivity Note:
Human, mouse, rat, bovine and canine.
Application Notes:
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.
Control:
A431 cell lysate, CHO cell lysate, foreskin fibroblasts, 3T3/A31 cells, L6 cells.
Included Positive Antigen Control: Catalog # 12-301, non stimulated A431 lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.
Included Positive Antigen Control: Catalog # 12-301, non stimulated A431 lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.
Quality Assurance:
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.
Purification Method:
Protein G purfied
Presentation:
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 Conditions:
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.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- EPHA2
- EphA2
- ECK
Alternate Names:
- 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
Usage Statement:
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.
Key Applications:
- Immunocytochemistry
- Immunoprecipitation
- Enzyme Assay
- Western Blotting
Entrez Gene Summary:
This gene belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. 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. This gene encodes a protein that binds ephrin-A ligands.
UniProt Summary:
FUNCTION: SwissProt: P29317 # Receptor for members of the ephrin-A family. Binds to ephrin-A1, -A3, -A4 and -A5.
SIZE: 976 amino acids; 108254 Da
SUBUNIT: Interacts with SLA (By similarity). Interacts with INPPL1/SHIP2.
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
TISSUE SPECIFICITY: Expressed most highly in tissues that contain a high proportion of epithelial cells, e.g., skin, intestine, lung, and ovary.
SIMILARITY: SwissProt: P29317 ## Belongs 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.
SIZE: 976 amino acids; 108254 Da
SUBUNIT: Interacts with SLA (By similarity). Interacts with INPPL1/SHIP2.
SUBCELLULAR LOCATION: Membrane; Single-pass type I membrane protein.
TISSUE SPECIFICITY: Expressed most highly in tissues that contain a high proportion of epithelial cells, e.g., skin, intestine, lung, and ovary.
SIMILARITY: SwissProt: P29317 ## Belongs 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.
Brand Family:
Upstate
Product Name:
Anti-Eck/EphA2, clone D7
Concentration:
1 mg/mL
Antibody Type:
Monoclonal Antibody
Qty/Pk:
200 µg
Format:
Purified
Host:
Mouse