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      Alexa Fluor™ 647 Biosimilar Anti-Human EGFR
      Alexa Fluor™ 647 Biosimilar Anti-Human EGFR
      Flow cytometric analysis of EGFR (Cetuximab Biosimilar) expression on Human A431 cells. Cells from the Human A431 (Cells from the Human epithelial carcinoma; ATCC® CRL-1555™) cell line were stained with either BD Pharmingen™ Alexa Fluor™ 647 Human IgG1, κ Isotype Control (Cat. No. 569961; Left Plot) or Alexa Fluor™ 647 Biosimilar Anti-Human EGFR antibody (Cat. No. 571603/571604; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The fluorescence histogram showing EGFR (Cetuximab Biosimilar) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Alexa Fluor™ 647 Biosimilar Anti-Human EGFR
      Flow cytometric analysis of EGFR (Cetuximab Biosimilar) expression on Human A431 cells. Cells from the Human A431 (Cells from the Human epithelial carcinoma; ATCC® CRL-1555™) cell line were stained with either BD Pharmingen™ Alexa Fluor™ 647 Human IgG1, κ Isotype Control (Cat. No. 569961; Left Plot) or Alexa Fluor™ 647 Biosimilar Anti-Human EGFR antibody (Cat. No. 571603/571604; Right Plot). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815) was added to cells right before analysis. The fluorescence histogram showing EGFR (Cetuximab Biosimilar) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Pharmingen™
      EGFR; ERBB1; HER1
      Human (QC Testing)
      Human IgG1, κ
      Human A431 Cell Line
      Flow cytometry (Routinely Tested)
      5 µl/test
      1956
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      4. 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.
      5. 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).
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. An isotype control should be used at the same concentration as the antibody of interest.
      8. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      11. For U.S. patents that may apply, see bd.com/patents.
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      571603 Rev. 1
      Antibody Details
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      Cetuximab297.rMAb

      The Cetuximab297.rMAb is a research grade, chimeric recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of Epidermal Growth Factor Receptor (EGFR) which is also known as HER1 or ErbB-1. The Cetuximab.rMAb is a biosimilar antibody that uses the same variable region sequences as the chimeric (mouse/human) therapeutic antibody, Cetuximab, combined with constant region sequences derived from a consensus sequence of human IgG1, kappa. The asparagine at position 297 of the constant heavy chain has been replaced with alanine (N297A) to further reduce Fc receptor interactions of this reagent. Cetuximab was derived from the Mouse 225 antibody that specifically binds to the extracellular domain of human EGFR and blocks its signaling activity.

         EGFR is a ~170 kDa single pass type I transmembrane glycoprotein that is encoded by EGFR which belongs to the HER/ErbB family of receptor tyrosine kinases (RTKs) that includes HER2 (ErbB-2, Neu, CD340), HER3 (ErbB-3), and HER4 (ErbB-4). EGFR is expressed on various cells including epithelial and endothelial cells. It plays and important role in their proliferation, survival, and motility in response to various ligands including EGF, heparin-binding EGF, amphiregulin, betacellulin, epiregulin, vaccinia virus growth factor and TGF-α. Upon ligand binding, EGFR undergoes homodimerization or heterodimerization with the other ErbB family members. This induces EGFR transautophosphorylation and stimulation of numerous downstream intracellular signaling pathways including the RAS/RAF/MEK/ERK, PLC-γ/PKC, PI3K/AKT, JAK/STAT, and NF-κB pathways. Activated EGFR-ligand complexes are removed from the cell surface via endocytosis and are degraded in the lysosomes to attenuate the signaling. Aberrant EGFR expression and signaling has been reported in inflammatory and malignant diseases.

         The Cetuximab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

      571603 Rev. 1
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-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.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      571603 Rev.1
      Citations & References
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      View product citations for antibody "571603" on CiteAb

      Development References (6)

      1. Fan Z, Lu Y, Wu X, Mendelsohn J. Antibody-induced epidermal growth factor receptor dimerization mediates inhibition of autocrine proliferation of A431 squamous carcinoma cells.. J Biol Chem. 1994; 269(44):27595-602. (Biology). View Reference
      2. Goldstein NI, Prewett M, Zuklys K, Rockwell P, Mendelsohn J. Biological efficacy of a chimeric antibody to the epidermal growth factor receptor in a human tumor xenograft model.. Clin Cancer Res. 1995; 1(11):1311-8. (Biology). View Reference
      3. Matar P, Rojo F, Cassia R, et al. Combined epidermal growth factor receptor targeting with the tyrosine kinase inhibitor gefitinib (ZD1839) and the monoclonal antibody cetuximab (IMC-C225): superiority over single-agent receptor targeting.. Clin Cancer Res. 2004; 10(19):6487-501. (Biology). View Reference
      4. Patel D, Lahiji A, Patel S, et al. Monoclonal antibody cetuximab binds to and down-regulates constitutively activated epidermal growth factor receptor vIII on the cell surface.. Anticancer Res. 2007; 27(5A):3355-66. (Biology). View Reference
      5. Sato JD, Kawamoto T, Le AD, Mendelsohn J, Polikoff J, Sato GH. Biological effects in vitro of monoclonal antibodies to human epidermal growth factor receptors.. Mol Biol Med. 1983; 1(5):511-29. (Biology). View Reference
      6. Wee P, Wang Z. Epidermal Growth Factor Receptor Cell Proliferation Signaling Pathways.. Cancers (Basel). 2017; 9(5):52. (Biology). View Reference
      View All (6) View Less
      571603 Rev. 1

      Please refer to Support Documents for Quality Certificates

       

      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.