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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 TH9 antibody monoclonal antibody specifically binds to CD59, a 21 kDa glycosyl-phosphatidyl inositol-anchored cell-surface glycoprotein of the Ly-6 superfamily. CD59 is expressed by many types of non-hematopoietic cells. In the rat hematopoietic system, CD59 has been detected on erythrocytes, monocytes, and some lymphocytes, but not on platelets. Soluble CD59 is found in body fluids and urine. CD59 is a complement regulatory protein that acts late in the complement cascade to prevent formation of the membrane attack complex (MAC). Therefore, CD59 is one of several proteins whose function is to protect host tissue from complement attack. Rat CD59 binds rat and human complement components and inhibits cytolysis mediated by complement from multiple species. CD59 has also been suggested to be a ligand for CD2 and to participate in T-cell costimulation.
Development References (6)
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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. (Biology). View Reference
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Hughes TR, Piddlesden SJ, Williams JD, Harrison RA, Morgan BP. Isolation and characterization of a membrane protein from rat erythrocytes which inhibits lysis by the membrane attack complex of rat complement. Biochem J. 1992; 284(1):169-176. (Immunogen: ELISA, Enhancement, Flow cytometry, Functional assay, Western blot). View Reference
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Lehto T, Morgan BP, Meri S. Binding of human and rat CD59 to the terminal complement complexes. Immunology. 1997; 90(1):121-128. (Biology). View Reference
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Liversidge J, Dawson R, Hoey S, McKay D, Grabowski P, Forrester JV. CD59 and CD48 expressed by rat retinal pigment epithelial cells are major ligands for the CD2-mediated alternative pathway of T cell activation. J Immunol. 1996; 156(10):3696-3703. (Clone-specific: (Co)-stimulation, Stimulation). View Reference
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Rushmere NK, Tomlinson S, Morgan BP. Expression of rat CD59: functional analysis confirms lack of species selectivity and reveals that glycosylation is not required for function. Immunology. 1997; 90(4):640-646. (Biology). View Reference
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Sugita Y, Masuho Y. CD59: its role in complement regulation and potential for therapeutic use. Immunotechnology. 1995; 1(3-4):157-168. (Biology). 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.