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BV605 Mouse Anti-Human C3b/iC3b
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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
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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  10. CF™ is a trademark of Biotium, Inc.
  11. 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 BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  12. BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
752742 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.

This antibody is conjugated to BD Horizon™ BV605 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max of 602-nm, BD Horizon BV605 can be excited by a violet laser and detected with a standard 610/20-nm filter set. BD Horizon BV605 is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em max at 605-nm. Due to the excitation of the acceptor dye by the green (532 nm) and yellow-green (561 nm) lasers, there will be significant spillover into the PE and BD Horizon PE-CF594 detectors off the green or yellow-green lasers. BD Horizon BV605 conjugates are very bright, often exhibiting brightness equivalent to PE conjugates and can be used as a third color off of the violet laser.

752742 Rev. 1
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV605
Violet 405 nm
407 nm
605 nm
752742 Rev.1
Citations & References
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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):1010-1019. (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):3280-3288. (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):1071-1079. (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):551-562. (Immunogen: Cell separation, Radioimmunoassay). View Reference
View All (6) View Less
752742 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.