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      Alexa Fluor™ 647 Biosimilar Anti-Human TNF

      BD Pharmingen™ Alexa Fluor™ 647 Biosimilar Anti-Human TNF

      Clone Adalimumab297.rMAb (also known as Adalimumab N297A Biosimilar.rMAb)

      (RUO)
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      Alexa Fluor™ 647 Biosimilar Anti-Human TNF
      Multiparameter flow cytometric analysis of TNF (Adalimumab Biosimilar)  expression by stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either Alexa Fluor™ 647 Human IgG1 (N297A), κ Isotype Control (Cat No. 569794; Left Plot) or Alexa Fluor™ 647 Biosimilar Anti-Human TNF (Adalimumab N297A Biosimilar) [Cat No. 569793/569795; Right Plot]. The pseudocolor density plot showing the correlated expression of TNF (Adalimumab Biosimilar) [or Ig Isotype control staining] versus forward light-scatter (FSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
      Alexa Fluor™ 647 Biosimilar Anti-Human TNF
      Multiparameter flow cytometric analysis of TNF (Adalimumab Biosimilar)  expression by stimulated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (Sigma P-8139; 50 ng/ml final concentration) and Ionomycin (Sigma I-0634; 1 μg/ml final concentration) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either Alexa Fluor™ 647 Human IgG1 (N297A), κ Isotype Control (Cat No. 569794; Left Plot) or Alexa Fluor™ 647 Biosimilar Anti-Human TNF (Adalimumab N297A Biosimilar) [Cat No. 569793/569795; Right Plot]. The pseudocolor density plot showing the correlated expression of TNF (Adalimumab Biosimilar) [or Ig Isotype control staining] versus forward light-scatter (FSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
      Product Details
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      BD Pharmingen™
      TNF; TNF-a; TNF-alpha; TNF-α; TNFA; TNFSF2; tumor necrosis factor-alpha
      Human (QC Testing)
      Human IgG1, κ
      Human TNF
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      7124
      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 the dye under optimum conditions and unreacted dye was removed.
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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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      569795 Rev. 1
      Antibody Details
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      Adalimumab297.rMAb

      The Adalimumab297.rMAb (also known as, Adalimumab N297A Biosimilar.rMAb) is a research grade recombinant Human IgG1, kappa monoclonal antibody that specifically recognizes Human Tumor Necrosis Factor (TNF) similarly to therapeutic Adalimumab. Adalimumab297.rMAb was engineered to code for a replacement of asparagine with alanine at position 297 (N297A) of the Human IgG1 heavy chain to reduce potential nonspecific staining caused by Fc receptor binding. Adalimumab is a recombinant Human IgG1 monoclonal anti-Human TNF antibody that is used in the treatment of a wide variety of inflammatory conditions such as rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, septic shock, and cancer. Therapeutic Adalimumab specifically binds to TNF and neutralizes its biological activity by blocking its binding to its cell surface receptors. Adalimumab does not bind to or inactivate lymphotoxin (Tumor necrosis factor-beta; TNF-β). TNF, also known as TNF alpha (TNF-α) or Cachectin, is an ~26 kDa type II transmembrane protein that is encoded by TNF. TNF is a multifunctional cytokine that plays roles in inflammation, immune system development, innate and adaptive immune responses, apoptosis, lipid metabolism, and necrosis of some tumor cells. TNF is variably produced by a wide variety of activated leucocytes, epithelial cells, endothelial cells, and tumor cells. This mediator is expressed as a noncovalently bound homotrimer that is expressed on the cell surface. Membrane TNF is enzymatically cleaved from the cell surface by tumor necrosis factor alpha converting enzyme (TACE), also known as ADAM17 and CD156b, into a soluble biologically active trimer of the TNF extracellular domains. TNF mediates its biological activities through binding to cell surface TNF Receptor Type 1 (TNFR1, CD120a) and Type 2 (TNFR2, CD120b), which are widely expressed on normal hemopoietic and nonhemopoietic cells as well as tumor cells.

         The Adalimumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

      569795 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
      569795 Rev.1
      Citations & References
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      View product citations for antibody "569795" on CiteAb

      Development References (4)

      1. Grewal IS. Overview of TNF superfamily: a chest full of potential therapeutic targets. Adv Exp Med Biol. 2009; 647:1-7. (Biology). View Reference
      2. Ingelman-Sundberg HM, Laestadius Å, Chrapkowska C, et al. Diverse effects on vaccine-specific serum IgG titres and memory B cells upon methotrexate and anti-TNF-α therapy in children with rheumatic diseases: A cross-sectional study.. Vaccine. 2016; 34(10):1304-11. (Biology). View Reference
      3. Meager A. Assays for cytotoxicity.. Methods Mol Biol. 2004; 249:135-52. (Methodology). View Reference
      4. Ward-Kavanagh LK, Lin WW, Šedý JR, Ware CF. The TNF Receptor Superfamily in Co-stimulating and Co-inhibitory Responses.. Immunity. 2016; 44(5):1005-19. (Biology). View Reference
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      569795 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.