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RY775 Human Anti-Human HER2

BD OptiBuild™ RY775 Human Anti-Human HER2

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

(RUO)
Product Details
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BD OptiBuild™
CD340; ERBB2; HER-2; HER-2/neu; HER2; NEU; proto-oncogene c-ErbB-2
Human (Tested in Development)
Human IgG1, κ
A431 Cell Line Expressing EGFR
Flow cytometry (Qualified)
0.2 mg/ml
2064
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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. An isotype control should be used at the same concentration as the antibody of interest.
  10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  12. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
  13. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
770289 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.

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Format Details
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RY775
The BD Horizon RealYellow™ 775 (RY775) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY775 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter).
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RY775
Yellow-Green 561 nm
557 nm
775 nm
770289 Rev.1
Citations & References
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View product citations for antibody "770289" 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
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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.