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Multiparameter flow cytometric analysis of CD222 expression by human leucocyte populations. Left Panel - Surface Staining: Human whole blood was stained with Alexa Fluor® 488 Mouse Anti-Human CD3 antibody (Cat. No. 557694) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Top Plots) or Alexa Fluor® 647 Mouse Anti-Human CD222 antibody (Cat. No. 565105; Bottom Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of CD222 (or Ig Isotype control staining) versus side light-scattered signals (SSC) or CD3 were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes (Left Plots) or lymphocytes (Right Plots), respectively. Right Panel - Surface and Intracellular Staining: Human whole blood was treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) to lyse erythrocytes and fix cells. The leucocytes were then permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723) and stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Top Plots) or Alexa Fluor® 647 Mouse Anti-Human CD222 antibody (Bottom Plots). Two-color flow cytometric dot plots showing the correlated expression of CD222 (or Ig Isotype control staining) versus side light-scattered signals (SSC) or CD3 were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes (Left Plots) or lymphocytes (Right Plots), respectively. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human CD222
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- 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.
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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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.