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      Alexa Fluor™ 488 Rat Anti-Mouse C1q
      Alexa Fluor™ 488 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 (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 Alexa Fluor™ 488 Rat IgG1, κ Isotype Control (Cat. No. 557720; dashed line histograms) or Alexa Fluor™ 488 Rat Anti-Mouse C1q antibody (Cat. No. 570987; solid line histograms) at 1 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 FACSCanto™ II Flow Cytometry System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Alexa Fluor™ 488 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 (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 Alexa Fluor™ 488 Rat IgG1, κ Isotype Control (Cat. No. 557720; dashed line histograms) or Alexa Fluor™ 488 Rat Anti-Mouse C1q antibody (Cat. No. 570987; solid line histograms) at 1 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 FACSCanto™ II Flow Cytometry System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      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 unreacted dye was 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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
      3. 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.
      4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. An isotype control should be used at the same concentration as the antibody of interest.
      7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      10. For U.S. patents that may apply, see bd.com/patents.
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      570987 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.

      570987 Rev. 1
      Format Details
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      Alexa Fluor™ 488
      Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 488
      Blue 488 nm
      494 nm
      517 nm
      570987 Rev.1
      Citations & References
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      View product citations for antibody "570987" 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
      570987 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.