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Alexa Fluor™ 488 Biosimilar Anti-Human HER2

BD Pharmingen™ Alexa Fluor™ 488 Biosimilar Anti-Human HER2

Clone Trastuzumab297.rMAb (also known as Trastuzumab N297A Biosimilar.rMAb)

(RUO)
Alexa Fluor™ 488 Biosimilar Anti-Human HER2
Flow cytometric analysis of HER2 (Trastuzumab Biosimilar) expression on viable Human SK-BR-3 cells. Cells from the Human SK-BR-3 (Breast adenocarcinoma, ATCC® HTB-30™) cell line were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564220) and then stained with either Alexa Fluor™ 488 IgG1, κ N297A Human Isotype Control (Cat. No. 570390; dashed line histogram) or Alexa Fluor™ 488 Biosimilar Anti-Human HER2 antibody (Cat. No. 570381/570451; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing HER2 (Trastuzumab Biosimilar) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
Flow cytometric analysis of HER2 (Trastuzumab Biosimilar) expression on viable Human SK-BR-3 cells. Cells from the Human SK-BR-3 (Breast adenocarcinoma, ATCC® HTB-30™) cell line were preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564220) and then stained with either Alexa Fluor™ 488 IgG1, κ N297A Human Isotype Control (Cat. No. 570390; dashed line histogram) or Alexa Fluor™ 488 Biosimilar Anti-Human HER2 antibody (Cat. No. 570381/570451; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing HER2 (Trastuzumab Biosimilar) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software.
Product Details
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BD Pharmingen™
CD340; ERBB2; HER-2; HER-2/neu; HER2; NEU; proto-oncogene c-ErbB-2
Human (QC Testing)
Human IgG1, κ
A431 Cell Line Expressing EGFR
Flow cytometry (Routinely Tested)
5 µl/test
2064
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.

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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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.
570381 Rev. 1
Antibody Details
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Trastuzumab297.rMAb

The Trastuzumab297.rMAb (also known as Trastuzumab N297A Biosimilar.rMAb) is a research grade, chimeric recombinant human IgG1 kappa antibody that specifically recognizes the extracellular domain of Human Epidermal Growth Factor Receptor 2 (also known as HER2, HER-2, ErbB2, NEU, or CD340) similarly to the therapeutic Trastuzumab antibody. The Trastuzumab297.rMAb uses the same variable region sequences as Trastuzumab, combined with constant region sequences derived from consensus sequences 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. Trastuzumab is a humanized therapeutic monoclonal antibody that was originally made by inserting the immunoglobulin (Ig) variable region sequences of the human HER-2-specific mouse monoclonal, 4D5. HER2 is a ~185 kDa type I transmembrane glycoprotein that is encoded by ERBB2 (erb-b2 receptor tyrosine kinase 2) which belongs to the ERBB family of receptor tyrosine kinases that includes EGFR (HER1), HER3, and HER4. Members of this receptor subfamily mediate the proliferation and differentiation of normal cells. Although HER2 has no identified direct-binding ligand, it forms signaling heterodimeric receptors with other ligand-bound ERBB family members. HER2 activates intracellular signaling pathways including MAPK and PI-3K through its cytoplasmic tyrosine kinase-containing region.  HER2 is expressed by bone marrow mesenchymal stem cells. It is overexpressed and plays a significant role in the development of multiple human cancers including some leukemias, neuroblastomas, and carcinomas.

      The Trastuzumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

570381 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
570381 Rev.1
Citations & References
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View product citations for antibody "570381" on CiteAb

Development References (3)

  1. Carter P, Presta L, Gorman CM, et al. Humanization of an anti-p185HER2 antibody for human cancer therapy.. Proc Natl Acad Sci U S A. 1992; 89(10):4285-9. (Biology). View Reference
  2. Goldenberg MM. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer.. Clin Ther. 1999; 21(2):309-18. (Biology). View Reference
  3. Stancovski I, Hurwitz E, Leitner O, Ullrich A, Yarden Y, Sela M. Mechanistic aspects of the opposing effects of monoclonal antibodies to the ERBB2 receptor on tumor growth.. Proc Natl Acad Sci USA. 1991; 88(19):8691-5. (Biology). View Reference
570381 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.