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      PE Mouse Anti-C3/C3b/iC3b
      PE Mouse Anti-C3/C3b/iC3b
      Flow cytometric analysis of human C3/C3b/iC3b deposition on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were treated with either Purified NA/LE Mouse Anti-Human CD3 antibody (Cat. No. 555336, Left Plot) or Purified NA/LE Mouse IgG1, κ Isotype Control (Cat. No. 554645, Right Plot) and were incubated with human serum (as a source of complement) for 30 min at 37°C. After washing, the cells were stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-C3/C3b/iC3b antibody (Cat. No. 567667; solid line histograms) at 0.5 µg/test. The fluorescence histograms showing the deposition of human C3/C3b/iC3b were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      PE Mouse Anti-C3/C3b/iC3b
      Flow cytometric analysis of mouse C3/C3b/iC3b deposition on C57BL/6 thymocytes. Thymocytes from a C57BL/6 mouse were treated with either Purified NA/LE Rat Anti-Mouse CD90.2 antibody (Cat. No. 553008, Left Plot) or Purified NA/LE Rat IgG2b, κ Isotype Control (Cat. No. 555845, Right Plot) and were incubated with mouse serum (as a source of complement) for 20 min at 4°C. After washing, the cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-C3/C3b/iC3b antibody (Cat. No. 567667; solid line histograms). The fluorescence histograms showing the deposition of mouse C3/C3b/iC3b were derived from gated events with the forward and side light-scatter characteristics of viable thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      PE Mouse Anti-C3/C3b/iC3b PE Mouse Anti-C3/C3b/iC3b
      Flow cytometric analysis of human C3/C3b/iC3b deposition on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were treated with either Purified NA/LE Mouse Anti-Human CD3 antibody (Cat. No. 555336, Left Plot) or Purified NA/LE Mouse IgG1, κ Isotype Control (Cat. No. 554645, Right Plot) and were incubated with human serum (as a source of complement) for 30 min at 37°C. After washing, the cells were stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-C3/C3b/iC3b antibody (Cat. No. 567667; solid line histograms) at 0.5 µg/test. The fluorescence histograms showing the deposition of human C3/C3b/iC3b were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Flow cytometric analysis of mouse C3/C3b/iC3b deposition on C57BL/6 thymocytes. Thymocytes from a C57BL/6 mouse were treated with either Purified NA/LE Rat Anti-Mouse CD90.2 antibody (Cat. No. 553008, Left Plot) or Purified NA/LE Rat IgG2b, κ Isotype Control (Cat. No. 555845, Right Plot) and were incubated with mouse serum (as a source of complement) for 20 min at 4°C. After washing, the cells were stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; dotted line histograms) or PE Mouse Anti-C3/C3b/iC3b antibody (Cat. No. 567667; solid line histograms). The fluorescence histograms showing the deposition of mouse C3/C3b/iC3b were derived from gated events with the forward and side light-scatter characteristics of viable thymocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Complement Component 3; C3; C3b; C3bi; iC3b
      Human (QC Testing), Mouse (Tested in Development)
      Mouse IgG1, κ
      Human C3
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      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 antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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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 cells 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. 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.
      3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      6. An isotype control should be used at the same concentration as the antibody of interest.
      7. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
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      567667 Rev. 1
      Antibody Details
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      6C9

      The 6C9 monoclonal antibody specifically recognizes human and mouse Complement component 3 (C3) and its multifunctional C3b and iC3b fragments. C3 is a ~190 kDa plasma glycoprotein that functions as a key component of the classical, alternative and lectin complement activation pathways. Active C3 fragments participate in innate and adaptive immune responses that eliminate pathogens, dying cells, and immune complexes from the body. C3 is produced by hepatocytes, mast cells, and leucocytes including some monocytes, macrophages, dendritic cells (DC), and T cells. As a result of complement activation, eg, initiated by an antigen-antibody immune complex in the classical pathway, biologically active C3b fragments can be cleaved from C3 by enzymatic C3 convertases. C3b can function as an opsonin by binding covalently via its reactive thioester to the surfaces of target cells, including microbes. C3b can also bind to antigen-antibody immune complexes. Macrophages and neutrophils can recognize and bind to C3b by their complement receptors, such as Complement Receptor 1 (CR1, CD35), which enhances their phagocytosis of C3b-bound target cells or immune complexes. Opsonization of target surfaces, including the cell surfaces of pathogenic organisms, infected or tumor cells, through C3b deposition is central to all three pathways of complement activation during innate or adaptive immune responses. C3b can also associate with other components of the complement system to form a C5 convertase. This complex cleaves C5 into the C5a anaphylatoxin and C5b which initiates formation of the of the cytolytic membrane attack complex (MAC) that can form holes in target cell membranes leading to cell lysis. C3b can be further cleaved into iC3b by factor I protease and its cofactors. The iC3b fragment serves as a ligand for engaging lymphoid and phagocytic cells via various receptors including CR2 (CD21), CR3 (CD11b/CD18), CR4 (CD11c/CD18), and CRIg.

              

      567667 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
      567667 Rev.1
      Citations & References
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      View product citations for antibody "567667" on CiteAb

      Development References (5)

      1. Colonna L, Parry GC, Panicker S, Elkon KB. Uncoupling complement C1s activation from C1q binding in apoptotic cell phagocytosis and immunosuppressive capacity.. Clin Immunol. 2016; 163:84-90. (Clone-specific: Flow cytometry). View Reference
      2. Dufloo J, Guivel-Benhassine F, Buchrieser J, et al. Anti-HIV-1 antibodies trigger non-lytic complement deposition on infected cells. EMBO Reports. 2020; 21(2):e49351. (Clone-specific: Flow cytometry). View Reference
      3. Lubbers R, van Essen MF, van Kooten C, Trouw LA. Production of complement components by cells of the immune system.. Clin Exp Immunol. 2017; 188(2):183-194. (Biology). View Reference
      4. Shi J, Rose EL, Singh A, et al. TNT003, an inhibitor of the serine protease C1s, prevents complement activation induced by cold agglutinins.. Blood. 2014; 123(26):4015-22. (Clone-specific: Flow cytometry). View Reference
      5. Sun D, Zhang M, Liu G, et al. Real-Time Imaging of Interactions of Neutrophils with Cryptococcus neoformans Demonstrates a Crucial Role of Complement C5a-C5aR Signaling.. Infect Immun. 2016; 84(1):216-29. (Clone-specific: Flow cytometry). View Reference
      View All (5) View Less
      567667 Rev. 1

       

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