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Flow cytometric analysis of CD172a/b expression on human peripheral blood monocytes. Whole blood was stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human CD172a/b antibody (Cat. No. 565035; solid line histogram). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD172a/b expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact monocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Pharmingen™ Alexa Fluor® 647 Mouse Anti-Human CD172a/b
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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
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The SE5A5 monoclonal antibody specifically binds to a common epitope on CD172a/SIRPα (90 kDa) and CD172b/SIRPβ1 (50 kDa). These transmembrane glycoproteins are members of the Signal Regulatory Protein (SIRP) family that, in turn, belongs to the Immunoglobulin superfamily. The SIRP family is comprised of two subgroups, SIRPα and SIRPβ that are distinguished by the presence (α) or absence (β) of a cytoplasmic domain containing two immunoreceptor tyrosine-based inhibition motifs (ITIM). CD172a/SIRPα is expressed on CD34+ stem/progenitor cells, cardiomyocytes, monocytes, macrophages, granulocytes, dendritic cells, and in the central nervous system. It binds to CD47 and is implicated in mediating inhibitory signals via the ITIM/SHP-2 association. CD172b/SIRPβ1 does not possess a cytoplasmic domain but instead the transmembrane domain contains a positively-charged residue that can interact with another transmembrane protein, DAP-12/KARAP. DAP-12 has two immunoreceptor tyrosine-based activation motifs (ITAM) within its cytoplasmic domain that are thought to link CD172b to cellular activation signaling. CD172b is expressed on myeloid cells, including peripheral blood monocytes and granulocytes. It is not expressed on CD34+ cells. CD172a and CD172b have complementary roles in signal regulation and may work together in tuning certain cellular responses to stimuli.
Development References (7)
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Bühring HJ, Simmons DL, Vernon-Wilson E. Review—CD172—SIRP; signal regulatory protein. 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; 2002:35.
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Dietrich J, Cella M, Seiffert M, Bühring HJ, Colonna M. Cutting edge: signal-regulatory protein beta 1 is a DAP12-associated activating receptor expressed in myeloid cells. J Immunol. 2000; 164(1):9-12. (Biology). View Reference
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Dubois NC, Craft AM, Sharma P, et al. SIRPA is a specific cell-surface marker for isolating cardiomyocytes derived from human pluripotent stem cells. Nat Biotechnol. 2011; 29:1011-1018. (Biology). View Reference
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Ghannadan M, Hauswirth AW, Schernthaner GH, et al. Detection of novel CD antigens on the surface of human mast cells and basophils. Int Arch Allergy Immunol. 2002; 127(4):299-307. (Biology). View Reference
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Seiffert M, Brossart P, Cant C, et al. Signal-regulatory protein alpha (SIRPalpha) but not SIRPbeta is involved in T-cell activation, binds to CD47 with high affinity, and is expressed on immature CD34(+)CD38(-) hematopoietic cells.. Blood. 2001; 97(9):2741-9. (Clone-specific: Immunoprecipitation, Inhibition). View Reference
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Seiffert M, Cant C, Chen Z, et al. Human signal-regulatory protein is expressed on normal, but not on subsets of leukemic myeloid cells and mediates cellular adhesion involving its counterreceptor CD47. Blood. 1999; 94(11):3633-3643. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
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Simmons DL, Vernon-Wilson E. Structure and function of the signal regulatory proteins (SIRPs). 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; 2002:35-38.
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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.