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APC Mouse Anti-Human EphB2
APC Mouse Anti-Human EphB2
Flow cytometric analysis of EphB2 expression on human Colo 205 cells.   Cells from the human COLO 205 (Colorectal adenocarcinoma, ATCC CCL-222) cell line were stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 550854; dashed line histogram) or APC Mouse Anti-Human EphB2 antibody (Cat. No. 564699; solid line histogram). The fluorescence histograms showing EphB2 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of EphB2 expression on human Colo 205 cells.   Cells from the human COLO 205 (Colorectal adenocarcinoma, ATCC CCL-222) cell line were stained with either APC Mouse IgG1, κ Isotype Control (Cat. No. 550854; dashed line histogram) or APC Mouse Anti-Human EphB2 antibody (Cat. No. 564699; solid line histogram). The fluorescence histograms showing EphB2 expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
EPHB2; EphB2R; TYRO5; CAPB; DRT; EK5; EPHT3; EPTH3; HEK5; PCBC
Human (QC Testing), Mouse (Reported)
Mouse BALB/c IgG1, κ
Human EphB2 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
2048
AB_2738898
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564699 Rev. 1
Antibody Details
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2H9

The 2H9 monoclonal antibody specifically binds to the Ephrin Type-B Receptor 2 (EphB2).  EphB2 is a type I transmembrane glycoprotein that belongs to the Eph receptor family of tyrosine kinase receptors. EphB2 serves as a cell surface receptor tyrosine kinase for membrane-anchored ligands referred to as type B ephrins (ephrin-B). The EphB2 receptor can bind to ephrin-B1, ephrin-B2, and ephrin-B3. Transmembrane ephrin-B family members are key regulators of embryogenesis including development of the nervous and vascular systems.  The EphB2 receptor functions as a chemodirectant in regulating cellular migration. EphB2/ephrin-B interactions orchestrate cell positioning by regulating cellular adhesion and repulsion during development, thereby influencing cell fate, morphogenesis and organogenesis. Signaling can occur in a forward pathway when the EphB2 receptor tyrosine kinase is activated by bound ligand and in a reverse pathway when transmembrane ephrin-B ligands are activated by EphB2 receptor-mediated crosslinking.  In the adult body, Eph receptor signaling plays major roles in regulating the architecture and physiology of different tissues under normal as well as disease conditions such as cancer. Ephrin-B1 and ephrin-B2 levels are upregulated in the vasculature during inflammation. Ephrin-B2 molecules that are localized to the luminal endothelial surface can signal through the EphB2 which is expressed by monocytes. This interaction promotes monocyte differentiation into proinflammatory macrophages. In the intestinal epithelium, EphB2/ephrin-B interactions regulate both cell positioning and tumor progression. The differential expression patterns of EphB2 allows for the detection and isolation of various intestinal epithelial cell types. These include intestinal stem cells (ISCs) which express high levels of EphB2. The 2H9 antibody reportedly blocks the interaction of EphB2 with ephrin ligands and crossreacts with mouse EphB2.

564699 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
APC
Red 627-640 nm
651 nm
660 nm
564699 Rev.1
Citations & References
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Development References (6)

  1. Foster KE, Gordon J, Cardenas K, et al. EphB-ephrin-B2 interactions are required for thymus migration during organogenesis.. Proc Natl Acad Sci USA. 2010; 107(30):13414-9. (Biology). View Reference
  2. Jung P, Sato T, Merlos-Suárez A, et al. Isolation and in vitro expansion of human colonic stem cells. Nat Med. 2011; 17(10):1225-1227. (Biology). View Reference
  3. Liu H, Devraj K, Möller K, Liebner S, Hecker M, Korff T. EphrinB-mediated reverse signalling controls junctional integrity and pro-inflammatory differentiation of endothelial cells. Thromb Haemost. 13(112)(Biology). View Reference
  4. Mao W, Luis E, Ross S, et al. EphB2 as a therapeutic antibody drug target for the treatment of colorectal cancer. Cancer Res. 204; 64(3):781-788. (Immunogen: Blocking, Cytotoxicity, Depletion, ELISA, Flow cytometry, Fluorescence microscopy, Functional assay, Immunofluorescence, Inhibition, In vivo exacerbation, Neutralization, Radioimmunoassay). View Reference
  5. Merlos-Suárez A, Barriga FM, Jung P et al. The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse. Cell Stem Cell. 2011; 8(5):511-524. (Clone-specific: Flow cytometry). View Reference
  6. Pasquale EB. The Eph family of receptors. Curr Opin Cell Biol. 1997; 9(5):608-615. (Biology). View Reference
View All (6) View Less
564699 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.