BD Pharmingen™ Alexa Fluor™ 647 Mouse Anti-Mouse C1q
Clone JL-1 (RUO)
Flow cytometric analysis of Mouse C1q deposition on BALB/c thymocytes labelled with CD90.2 (Thy-1.2)-specific antibody followed by incubation with Mouse serum. BALB/c Mouse thymocytes were labeled 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 4°C. After washing, the cells were stained with either Alexa Fluor™ 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-Mouse C1q antibody (Cat. No. 571192/571193; solid line histograms) at 0.5 µg per test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to the cells right before analysis. The fluorescence histograms showing the deposited levels of Mouse C1q were derived from gated events with the 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.
Flow cytometric analysis of Mouse C1q deposition on BALB/c thymocytes labelled with CD90.2 (Thy-1.2)-specific antibody followed by incubation with Mouse serum. BALB/c Mouse thymocytes were labeled 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 4°C. After washing, the cells were stained with either Alexa Fluor™ 647 Mouse IgG2b, κ Isotype Control (Cat. No. 565378; dashed line histograms) or Alexa Fluor™ 647 Mouse Anti-Mouse C1q antibody (Cat. No. 571192/571193; solid line histograms) at 0.5 µg per test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to the cells right before analysis. The fluorescence histograms showing the deposited levels of Mouse C1q were derived from gated events with the 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.
Preparation And Storage
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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The JL-1 monoclonal antibody specifically recognizes the C1q component of the 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 follicular dendritic cells, 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. The JL-1 antibody recognizes the collagen-like region (CLR) of C1q and although generated against mouse C1q, reportedly crossreacts with human C1q.
Development References (3)
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Holers VM. Anti-C1q autoantibodies amplify pathogenic complement activation in systemic lupus erythematosus.. J Clin Invest. 2004; 114(5):616-9. (Clone-specific). View Reference
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Ling GS, Crawford G, Buang N, et al. C1q restrains autoimmunity and viral infection by regulating CD8(+) T cell metabolism. Science. 2018; 360(6388):558-563. (Clone-specific: Flow cytometry).
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Trouw LA, Groeneveld TW, Seelen MA, et al. Anti-C1q autoantibodies deposit in glomeruli but are only pathogenic in combination with glomerular C1q-containing immune complexes.. J Clin Invest. 2004; 114(5):679-88. (Immunogen: Blocking, Depletion, ELISA, Immunofluorescence, In vivo exacerbation, Western blot). View Reference
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