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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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The 512 monoclonal antibody specifically recognizes a 65-kDa and 55-kDa molecules of the erythrocyte membrane, which are the rat counterpart of mouse Crry/p65, a membrane inhibitor of complement component C3 convertase. Crry/p65 is widely distributed, predominantly expressed on endothelial cells and all circulating cells. This molecule was also detected on immature hepatocytes, systemic endothelial cells, skin fibroblasts, bronchial epithelial cells, bile canaliculi, the Schwann sheath of peripheral nerve fibers, and ependymal cells. Mouse Crry/p65 uses the same mechanisms as human Decay-Accelerating Factor (DAF) and human Membrane Cofactor Protein (MCP) in inhibiting C3 and C5 convertases, but it is not homologous to either DAF or MCP. It has been proposed that Crry/p65 is the mouse genetic homologue of human CR1 (CD35).
Development References (5)
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Funabashi K, Okada N, Matsuo S, Yamamoto T, Morgan BP, Okada H. Tissue distribution of complement regulatory membrane proteins in rats. Immunology. 1994; 81(3):444-451. (Clone-specific: Immunohistochemistry). View Reference
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Kim YU, Kinoshita T, Molina H, et al. Mouse complement regulatory protein Crry/p65 uses the specific mechanisms of both human decay-accelerating factor and membrane cofactor protein. J Exp Med. 1995; 181(1):151-159. (Biology). View Reference
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Kurtz CB, O'Toole E, Christensen SM, Weis JH. The murine complement receptor gene family. IV. Alternative splicing of Cr2 gene transcripts predicts two distinct gene products that share homologous domains with both human CR2 and CR1. J Immunol. 1990; 144(9):3581-3591. (Biology). View Reference
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Li B, Sallee C, Dehoff M, Foley S, Molina H, Holers VM. Mouse Crry/p65. Characterization of monoclonal antibodies and the tissue distribution of a functional homologue of human MCP and DAF. J Immunol. 1993; 151(8):4295-4305. (Biology). View Reference
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Takizawa H, Okada N, Okada H. Complement inhibitor of rat cell membrane resembling mouse Crry/p65. J Immunol. 1994; 152(6):3032-3038. (Immunogen: Blocking, Western blot). View Reference
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.