General description
ColorWheel® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel® antibody with any ColorWheel® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel® technology requires both ColorWheel® antibody and ColorWheel® dye for Flow Cytometry application.
We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry. This product is Preservative-free, lyophilized product for enhanced stability and allow for ambient shipping and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency".
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Specificity
Clone F2-27 is a mouse monoclonal antibody that detects Ephrin type-A receptor 2 (EphA2).
Immunogen
KP-2, KP-3, SUIT-2, and MIAPaCa-2 cells.
Application
Quality Control Testing
Evaluated by Flow Cytometry in PC-3 cells.
Flow Cytometry Analysis: Staining of one million PC-3 cells was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1172, Anti-Human EphA2 (F2-27) ColorWheel® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control conjugated to Cat. No. CWDS-PE ColorWheel® Antibody-Ready Phycoerythrin (PE) Dye.
Tested Applications
Flow Cytometry Analysis: Staining of one million human peripheral blood lymphocytes was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1172, Anti-Human EphA2 (F2-27) ColorWheel® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel® Antibody-Ready Allophycocyanin (APC) Dye or a 1:1 mixture of an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control conjugated to Cat. No. CWDS-APC ColorWheel® Antibody-Ready Allophycocyanin (APC) Dye.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Compatibility
Requires ColorWheel® Dye (Sold Separately)
Target description
Ephrin type-A receptor 2 (UniProt: P29317; also known as EC: 2.7.10.1, Epithelial cell kinase, Tyrosine-protein kinase receptor ECK) is encoded by the EPHA2 (also known as ECK) gene (Gene ID: 1969) in human. Ephrin type-A receptor 2 (EphA2) is a homodimeric single-pass type I membrane protein that is synthesized with a signal peptide (aa 1-23), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 24-537), a short transmembrane domain (aa 538-558), and a cytoplasmic domain (aa 559-976). It is expressed in brain and glioma tissue and in glioma cell lines. It is also highly expressed in tissues that contain a high proportion of epithelial cells. Its levels are up-regulated by UV irradiation via a TP53-independent, MAPK-dependent mechanism. EphA2 serves as a receptor tyrosine kinase that binds promiscuously membrane-bound ephrin-A family ligands residing on adjacent cells, leading to contact-dependent bidirectional signaling into neighboring cells. It is involved in the regulation of migration, integrin-mediated adhesion, and in proliferation and differentiation of cells. EphA2 is phosphorylated on tyrosine residues upon binding and activation by EFNA1. Phosphorylation on tyrosine 588 and 594 is reported to be essential for binding VAV2 and VAV3 while phosphorylation on Tyr 735 and Tyr 930 are required for binding PI3-kinase p85 subunit. Overexpression of EphA2 is reported in murine models of skin tumors. Inflammatory cytokines are reported to promote RSK-dependent phosphorylation of EphA2 at serine 897 and this RSK-EphA2 axis might contribute to cell motility and promote tumor malignant progression. Two isoforms of Ephrin type-A receptor 2 have been described that are produced by alternative splicing. This ColorWheel® Dye-Ready monoclonal antibody was developed using our Cat. No. MABS2026; Anti- Anti-EphA2, clone F2-27. (Ref.: Zhou, Y., et al. (2015). Nat. Commun. 6; 7679; Guo, H., et al. (2006). Cancer Res. 66(14); 7050-7058).
Physical form
Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.
Reconstitution
1.0 mg/mL after reconstitution at 25 µL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Storage and Stability
Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 µL PBS. Protect from light.
Legal Information
ColorWheel 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.