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      PE Rat Anti-Human IL-6
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      This SKU will be discontinuing Apr 2024. Suggested alternate SKU is [554697] or for additional support, contact your local applications specialist. Contact Us #
      PE Rat Anti-Human IL-6
      Expression of IL-6 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 6 hours with LPS (100 ng/ml final concentration) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC-mouse anti-human CD14 antibody (FITC-M5E2, Cat. No. 555397) and 0.25 µg of PE-rat anti-human IL-6 antibody (PE-MQ2-13A5, Cat. No. 554545) by using Pharmingen's staining protocol (left panel). The data reflect gating on monocytes, based on forward and side-scatter light signals. The binding of PE-MQ2-13A5 was blocked by the preincubation of the mouse antibody conjugate with an excess of recombinant human IL-6 (1 µg; Cat. 550071; right panel). The quadrant markers for the bivariate dot plots were set based on autofluorescence and verified using this ligand-blocking specificity control.
      PE Rat Anti-Human IL-6
      Expression of IL-6 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 6 hours with LPS (100 ng/ml final concentration) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with FITC-mouse anti-human CD14 antibody (FITC-M5E2, Cat. No. 555397) and 0.25 µg of PE-rat anti-human IL-6 antibody (PE-MQ2-13A5, Cat. No. 554545) by using Pharmingen's staining protocol (left panel). The data reflect gating on monocytes, based on forward and side-scatter light signals. The binding of PE-MQ2-13A5 was blocked by the preincubation of the mouse antibody conjugate with an excess of recombinant human IL-6 (1 µg; Cat. 550071; right panel). The quadrant markers for the bivariate dot plots were set based on autofluorescence and verified using this ligand-blocking specificity control.
      Product Details
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      BD Pharmingen™
      Human (QC Testing)
      Rat IgG1
      Human IL-6 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      AB_395469
      Aqueous buffered solution containing ≤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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      Recommended Assay Procedure:

      Immunofluorescent Staining For Intracellular Cytokines: The MQ2-13A5 antibody is useful for immunofluorescent staining and

      flow cytometric analysis to identify and enumerate IL-6 producing cells within mixed cell populations. The PE-conjugated MQ2-13A5 antibody (Cat. No. 554545) is especially suitable for these studies. The use of a specificity control, such as one of the following, is suggested: 1) recombinant human IL-6 (Cat. No. 550071), or 2) rat IgG1 isotype control, PE-R3-34 (Cat. No. 554685). For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook.

      ELISA Capture: The purified MQ2-13A5 antibody (Cat. No. 554543) is useful as a capture antibody for a sandwich ELISA for measuring human IL-6 protein levels. Purified MQ2-13A5 antibody can be paired with the biotinylated MQ2-39C3 antibody (Cat. No. 554546) as the detecting antibody, with recombinant human IL-6 protein (Cat. No. 550071) as the standard. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.

      Note: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring human IL-6 in serum or plasma our human IL-6 BD OptEIA™ set (Cat. No. 555220) or BD OptEIA kit (Cat. No. 550799) are specially formulated and recommended.

      Neutralization: The MQ2-13A5 antibody (Cat. No. 554541) is useful for neutralization of human IL-6 bioactivity.

      IP/WB: The MQ2-13A5 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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.
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      554545 Rev. 1
      Antibody Details
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      MQ2-13A5

      The MQ2-13A5 monoclonal antibody specifically binds to human interleukin-6 (IL-6). IL-6 is a multifunctional cytokine that plays a central role in host defense mechanisms, including hematopoiesis, immune responses (eg, T and B cell activation and differentiation) and acute phase reactions. IL-6 can be expressed by a variety of cells including monocytes/macrophages, eosinophils, fibroblasts, vascular endothelial cells, bone marrow stromal cells, mesangial cells, hepatocytes, keratinocytes, astrocytes, T lymphocytes, B lymphocytes, and various tumor cells. IL-6 production is upregulated by cells in response to bacterial products such as lipopolysaccharide, viruses and other pro-inflammatory cytokines such as IL-1, TNF , and IFN-γ. IL-6 transcription is downregulated by cells in response to IL-4 and IL-10. The functional IL-6 Receptor (IL-6R) complex consists of two transmembrane glycoproteins, an 80-kDa low-affinity ligand-binding receptor subunit (IL-6Rα/CD126) and a 130 kDa (gp130/CD130) subunit that binds to IL-6-IL-6Rα to form the high-affinity signal transducing complex. Abnormal expression of IL-6 is related to the pathogenesis of many diseases including neoplastic (eg, multiple myeloma) and autoimmune diseases (eg, rheumatoid arthritis). The immunogen used to generate this hybridoma was COS-7 -expressed recombinant human IL-6.

      This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

      554545 Rev. 1
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      554545 Rev.1
      Citations & References
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      View product citations for antibody "554545" on CiteAb

      Development References (4)

      1. Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA, Neutralization). View Reference
      2. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA, Neutralization). View Reference
      3. Gaines Das RE, Poole S. The international standard for interleukin-6. Evaluation in an international collaborative study. J Immunol Methods. 1993; 160(2):147-153. (Clone-specific: ELISA, Neutralization). View Reference
      4. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). View Reference
      View All (4) View Less
      554545 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.