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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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Companion Products
The MEM-238 monoclonal antibody specifically binds to CD222, which is also known as the Insulin-like growth factor 2 Receptor (IGF2R, IGF-II Receptor), Cation-independent mannose 6-phosphate receptor (CIMPR), or Mannose-6 phosphate receptor (M6PR). CD222 is ubiquitously expressed by a variety of cell types as a cell surface type I transmembrane glycoprotein. However, in the course of receptor trafficking between the cell surface and intracellular compartments, the majority of CD222 is found within cells. Cell surface CD222 functions as a multifunctional receptor that binds to a large number of extracellular ligands including acid hydrolases, insulin-like growth factors, latent TGF-β, leukemia inhibitory factor (LIF), proliferin, prorenin, plasminogen, and Herpes simplex virus. It regulates extracellular Insulin-like growth factor II (IGF-II/IGF-2) levels by binding and internalizing the growth factor for lysosomal degradation. This effectively removes IGF-II from the circulation and tissues and thus prevents it from signaling through the growth-stimulatory Insulin-like growth factor I Receptor (IGF-1R, CD221) pathway. However, studies have reported that IGF-II may activate some cellular functions through CD222 as well. A soluble form of the CD222 extracellular region can also be detected in human serum and may play a role in regulating IGF-II activity. CD222 serves as a surface receptor for latent TGFβ and can complex with plasminogen and CD87, a urokinase-type plasminogen activator receptor, to activate latent TGF-β. CD222 also binds to a variety of Mannose 6-phosphate (M6P)-containing proteins, including extracellular and newly-synthesized lysosomal enzymes, and transports them to lysosomes.
Development References (4)
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Brown J, Jones EY, Forbes BE. Keeping IGF-II under control: lessons from the IGF-II-IGF2R crystal structure. Trends Biochem Sci. 2009; 34(12):612-619. (Biology). View Reference
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Godar S, Leska V, Cebecauer M, Hilgert I, Horejsi V, Stockinger H. Cd222 (Mannose-6 phosphate/insulin-like growth factor II-receptor) Summary and Workshop report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:482-485.
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Leksa V, Godar S, Cebecauer M, et al. The N terminus of mannose 6-phosphate/insulin-like growth factor 2 receptor in regulation of fibrinolysis and cell migration. J Biol Chem. 2002; 277(43):40575-40582. (Immunogen: Flow cytometry, Western blot). View Reference
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Leksa V, Godar S, Schiller HB, et al. TGF-beta-induced apoptosis in endothelial cells mediated by M6P/IGFII-R and mini-plasminogen. J Cell Sci. 2005; 118(Pt19):4577-4586. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunoaffinity chromatography, Immunofluorescence, Immunoprecipitation, Western blot). View Reference
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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.