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      RB705 Mouse Anti-Human C3b/iC3b
      Product Details
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      BD OptiBuild™
      C3b; iC3b; C3bi; complement component C3b
      Human (Tested in Development)
      Mouse IgG1, κ
      Human C3b
      Flow cytometry (Qualified)
      0.2 mg/ml
      718
      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.
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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. 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. 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.
      8. 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.
      9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      10. An isotype control should be used at the same concentration as the antibody of interest.
      11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      12. 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.
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      757675 Rev. 1
      Antibody Details
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      3E7

      The 3E7 monoclonal antibody specifically recognizes multifunctional C3b and iC3b fragments of the ~190 kDa plasma glycoprotein, Complement component 3 (C3). C3 is a key component of the classical, alternative and lectin complement activation pathways which participate in innate and adaptive immune responses to 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 produce holes in target cell membranes. 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. The 3E7 antibody van reportedly block the activation of the alternative pathway of complement.

      757675 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      757675 Rev.1
      Citations & References
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      View product citations for antibody "757675" on CiteAb

      Development References (6)

      1. DiLillo DJ, Pawluczkowycz AW, Peng W, et al. Selective and efficient inhibition of the alternative pathway of complement by a mAb that recognizes C3b/iC3b.. Mol Immunol. 2006; 43(7):1010-9. (Immunogen: Functional assay, Inhibition). View Reference
      2. Kennedy AD, Beum PV, Solga MD, et al. Rituximab infusion promotes rapid complement depletion and acute CD20 loss in chronic lymphocytic leukemia.. J Immunol. 2004; 172(5):3280-8. (Clone-specific: Cytotoxicity, Immunofluorescence). View Reference
      3. Kennedy AD, Solga MD, Schuman TA, et al. An anti-C3b(i) mAb enhances complement activation, C3b(i) deposition, and killing of CD20+ cells by rituximab.. Blood. 2003; 101(3):1071-9. (Clone-specific: Flow cytometry). View Reference
      4. Lindorfer MA, Pawluczkowycz AW, Peek EM, Hickman K, Taylor RP, Parker CJ. A novel approach to preventing the hemolysis of paroxysmal nocturnal hemoglobinuria: both complement-mediated cytolysis and C3 deposition are blocked by a monoclonal antibody specific for the alternative pathway of complement.. Blood. 2010; 115(11):2283-91. (Clone-specific: Functional assay, Inhibition). View Reference
      5. 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
      6. Sokoloff MH, Nardin A, Solga MD, et al. Targeting of cancer cells with monoclonal antibodies specific for C3b(i).. Cancer Immunol Immunother. 2000; 49(10):551-62. (Immunogen: Cell separation, Radioimmunoassay). View Reference
      View All (6) View Less
      757675 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.