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      BV421 Rat Anti-Mouse C1q
      BV421 Rat Anti-Mouse C1q
      Flow cytometric analysis of antibody-dependent Mouse C1q deposition on Mouse thymocytes. BALB/c Mouse thymocytes were treated with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 555846, Left Plot) or Purified Rat Anti-Mouse CD90.2 antibody (Thy-1.2; Cat. No. 553009, Right Plot) for 20 min at 4°C. The cells were then incubated with Mouse serum (as a source of Complement) in the presence of 5 mM of EDTA, for 30 min at 37°C. After washing, the cells were stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; dashed line histograms) or BD Horizon™ BV421 Rat Anti-Mouse C1q antibody (Cat. No. 570367/570437; solid line histograms) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to the cells right before analysis. The fluorescence histograms showing the antibody-dependent deposition of Mouse C1q were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      BV421 Rat Anti-Mouse C1q
      Flow cytometric analysis of antibody-dependent Mouse C1q deposition on Mouse thymocytes. BALB/c Mouse thymocytes were treated with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 555846, Left Plot) or Purified Rat Anti-Mouse CD90.2 antibody (Thy-1.2; Cat. No. 553009, Right Plot) for 20 min at 4°C. The cells were then incubated with Mouse serum (as a source of Complement) in the presence of 5 mM of EDTA, for 30 min at 37°C. After washing, the cells were stained with either BD Horizon™ BV421 Rat IgG1, κ Isotype Control (Cat. No. 562868; dashed line histograms) or BD Horizon™ BV421 Rat Anti-Mouse C1q antibody (Cat. No. 570367/570437; solid line histograms) at 0.5 µg/test. DAPI Solution (Cat. No. 564907) was added to the cells right before analysis. The fluorescence histograms showing the antibody-dependent deposition of Mouse C1q were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Horizon™
      Complement C1q; Complement Component 1q; Complement protein C1q
      Mouse (QC Testing)
      Rat IgG1, κ
      Mouse C1q
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      12259, 12260, 12262
      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 to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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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.

         For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).

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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. 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.
      8. CF™ is a trademark of Biotium, Inc.
      9. For U.S. patents that may apply, see bd.com/patents.
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      570367 Rev. 1
      Antibody Details
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      RmC7H8

      The RmC7H8 monoclonal antibody specifically recognizes the C1q component of the mouse macromolecular C1 complex. The complex is comprised of C1q and 2 molecules each of the serine proteases, C1r and C1q. The C1 macromolecular complex, C1qC1r2C1s2, is bound together by Ca2+ ions. C1q is a serum protein that is synthesized by macrophages and microglia. It exists as an ~460 kDa protein formed from 18 polypeptide chains comprised of three different subunits named C1qa, C1qb, and C1qc. Each chain contains an N-terminal collagen-like sequence and a C-terminal globular gC1q module. Structural studies reveal that C1q is formed as a hexamer with 6 collagen-like triple helices, forming a central fiber bundle, each extended by globular domains. C1q is a multifunctional protein that can regulate a variety of cellular processes in addition to activating the classical complement pathway (CCP). C1q globular regions mediate target recognition such as binding to the Fc regions of IgG and IgM antibodies found in immune complexes including antibodies bound to pathogens or target cells and subsequent activation of the CCP. The C1q globular regions can also bind to bacterial and viral surface proteins as well as altered self elements including histones, DNA, and annexins on the surface of apoptotic or necrotic cells. The enhanced phagocytosis mediated through interaction of bound C1q with various receptors expressed by phagocytes, possibly combined with further complement activation and opsonization of cells, may contribute to the safe removal of stressed or dead cells that are pro-inflammatory. Conformational changes in target bound C1q's collagen-like regions can lead to interaction with and activation of the C1r and C1s proteases which result in activation of the CCP.

      570367 Rev. 1
      Format Details
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      BV421
      The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BV421
      Violet 405 nm
      407 nm
      423 nm
      570367 Rev.1
      Citations & References
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      View product citations for antibody "570367" on CiteAb

      Development References (8)

      1. Abbitt KB, Cotter MJ, Ridger VC, Crossman DC, Hellewell PG, Norman KE. Antibody ligation of murine Ly-6G induces neutropenia, blood flow cessation, and death via complement-dependent and independent mechanisms.. J Leukoc Biol. 2009; 85(1):55-63. (Clone-specific: Immunofluorescence). View Reference
      2. Ankeny DP, Guan Z, Popovich PG. B cells produce pathogenic antibodies and impair recovery after spinal cord injury in mice.. J Clin Invest. 2009; 119(10):2990-9. (Biology). View Reference
      3. Kang YS, Do Y, Lee HK, et al. A dominant complement fixation pathway for pneumococcal polysaccharides initiated by SIGN-R1 interacting with C1q.. Cell. 2006; 125(1):47-58. (Clone-specific: Functional assay). View Reference
      4. Libert C, Wielockx B, Grijalba B, et al. The role of complement activation in tumour necrosis factor-induced lethal hepatitis.. Cytokine. 1999; 11(8):617-25. (Clone-specific: Inhibition, In vivo exacerbation). 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. Schuh R, Kremmer E, Ego E, Wasiliu M, Thierfelder S. Determination of monoclonal antibody specificity by immunoadsorption and western blotting.. J Immunol Methods. 1992; 152(1):59-67. (Immunogen: ELISA, Immunoprecipitation). View Reference
      7. Thielens NM, Tedesco F, Bohlson SS, Gaboriaud C, Tenner AJ. C1q: A fresh look upon an old molecule. Mol Immunol. 2017; 89:73-83. (Biology). View Reference
      8. Zachrau B, Finke D, Kropf K, Gosink HJ, Kirchner H, Goerg S. Antigen localization within the splenic marginal zone restores humoral immune response and IgG class switch in complement C4-deficient mice.. Int Immunol. 2004; 16(12):1685-90. (Clone-specific: Immunohistochemistry). View Reference
      View All (8) View Less
      570367 Rev. 1

       

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      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.