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BV421 Mouse Anti-Human EphA2
BV421 Mouse Anti-Human EphA2
Flow cytometric analysis of EphA2 expression on human epidermoid carcinoma cells. A431 cells (ATCC CRL-1555) were harvested using trypsin, and then stained with either BD OptiBuild™ BV421 Mouse Anti-Human EphA2 (Cat. No. 748144, solid line histogram) or BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) at 0.25 µg/test. The fluorescence histogram showing EphA2 (or Ig Isotype control) staining was derived from gated events with the forward and side light-scatter characteristics of viable singlet cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo® software.   The above is qualification data only and does not represent a specific OptiBuild™ lot.
Flow cytometric analysis of EphA2 expression on human epidermoid carcinoma cells. A431 cells (ATCC CRL-1555) were harvested using trypsin, and then stained with either BD OptiBuild™ BV421 Mouse Anti-Human EphA2 (Cat. No. 748144, solid line histogram) or BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histogram) at 0.25 µg/test. The fluorescence histogram showing EphA2 (or Ig Isotype control) staining was derived from gated events with the forward and side light-scatter characteristics of viable singlet cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo® software.   The above is qualification data only and does not represent a specific OptiBuild™ lot.
Product Details
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BD OptiBuild™
EPHA2; ECK; ARCC2; CTPA; CTPP1; CTRCT6
Human (Tested in Development)
Mouse IgG2a, κ
Human EphA2 Recombinant Protein
Flow cytometry (Qualified)
0.2 mg/ml
AB_2872605
Aqueous buffered solution containing ≤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 with BD Horizon BV421 under optimal conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD OptiBuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  10. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
748144 Rev. 2
Antibody Details
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371805

The 371805 monoclonal antibody specifically recognizes Ephrin type-A receptor 2 (EphA2).  EphA2 is a ~130 kDa type I transmembrane glycoprotein that is encoded by EPHA2 (EPH receptor A2) which belongs to ephrin receptor subfamily within the receptor tyrosine kinase (RTK) family. EphA2 is also known as Epithelial cell kinase (ECK) or Epithelial cell receptor protein tyrosine kinase. The extracellular region of EphA2 contains an N-terminal globular domain, a cysteine-rich domain and 2 fibronectin type III repeats, whereas the cytoplasmic domain contains two tyrosine residues adjacent to the membrane that function as major autophosphorylation sites and a kinase domain. EphA2 is variably expressed by epithelial cells, dendritic cells, Langerhans cells, keratinocytes, and endothelial cells. EphA2 functions as a receptor for glycophosphatidylinositol (GPI) membrane-linked members of the Ephrin-A family, including Ephrins A1-A5. EphA2 is involved in regulating cellular growth, adhesion, migration, survival, and plays a role in angiogenesis. Its expression may be upregulated on vascular endothelium in certain breast, prostate, and colon cancers as well as on some metastatic tumor cells.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

748144 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
748144 Rev.2
Citations & References
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Development References (7)

  1. Brantley DM, Cheng N, Thompson EJ, et al. Soluble Eph A receptors inhibit tumor angiogenesis and progression in vivo. Oncogene. 2002; 21(46):7011. (Biology). View Reference
  2. Cheng N, Brantley DM, Liu H, et al. Blockade of EphA receptor tyrosine kinase activation inhibits vascular endothelial cell growth factor-induced angiogenesis. Mol Cancer Res. 2002; 1(1):2-11. (Biology). View Reference
  3. Liao-Chan S, Daine-Matsuoka B, Heald N, et al. Quantitative assessment of antibody internalization with novel monoclonal antibodies against Alexa fluorophores.. PLoS ONE. 2015; 10(4):e0124708. (Clone-specific: Bioassay). View Reference
  4. Lindberg RA, Hunter T. cDNA cloning and characterization of eck, an epithelial cell receptor protein-tyrosine kinase in the eph/elk family of protein kinases.. Mol Cell Biol. 1990; 10(12):6316-24. (Biology). View Reference
  5. Udayakumar D, Zhang G, Ji Z, Njauw CN, Mroz P, Tsao H. EphA2 is a critical oncogene in melanoma. Oncogene. 2011; 30(50):4921-4929. (Biology). View Reference
  6. Udyavar AR, Wooten DJ, Hoeksema M, et al. Novel Hybrid Phenotype Revealed in Small Cell Lung Cancer by a Transcription Factor Network Model That Can Explain Tumor Heterogeneity.. Cancer Res. 2017; 77(5):1063-1074. (Clone-specific: Flow cytometry). View Reference
  7. de Saint-Vis B, Bouchet C, Gautier G, Valladeau J, Caux C, Garrone P. Human dendritic cells express neuronal Eph receptor tyrosine kinases: role of EphA2 in regulating adhesion to fibronectin. Blood. 2003; 102(13):4431-4440. (Biology). View Reference
View All (7) View Less
748144 Rev. 2

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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.