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PerCP-Cy™5.5 Mouse Anti-Human CD193

BD Pharmingen™ PerCP-Cy™5.5 Mouse Anti-Human CD193

Clone 5E8 (also known as 5E8-G9-B4)

(RUO)
PerCP-Cy™5.5 Mouse Anti-Human CD193
Two-color flow cytometric analysis of CD193 expression on human peripheral blood granulocytes (eosinophils). Whole blood was stained with PerCP-Cy™5.5 Mouse Anti-Human CD193 antibody (Cat. No. 564189) and APC Mouse Anti-Human CD16 (Cat. No. 561248). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-color flow cytometric dot plot showing the correlated expression of CD16 versus CD193 was derived from events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Two-color flow cytometric analysis of CD193 expression on human peripheral blood granulocytes (eosinophils). Whole blood was stained with PerCP-Cy™5.5 Mouse Anti-Human CD193 antibody (Cat. No. 564189) and APC Mouse Anti-Human CD16 (Cat. No. 561248). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A two-color flow cytometric dot plot showing the correlated expression of CD16 versus CD193 was derived from events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
CCR3: Chemokine (C-C motif) receptor 3; C-C CKR-3; CKR3; CMKBR3
Human (QC Testing)
Mouse C57BL/6 IgG2b, κ
Human CCR3 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
1232
AB_2738656
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. 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.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of GE Healthcare.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564189 Rev. 3
Antibody Details
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5E8

The 5E8 monoclonal antibody specifically binds to human CCR3 which is also known as CD193. CCR3 is a G protein-linked, 7 transmembrane, chemokine receptor expressed on a variety of hematopoietic cells. Similar to CCR5 and CXCR4, CCR3 can be a co-receptor for HIV-1. It is primarily expressed by eosinophils and basophils during atopic conditions, dermatitis, allergic rhinitis, conjunctivitis and bronchial asthma. Chemokines including RANTES, Eotaxin, MCP-3, MIP1α have been reported to act as ligands for CCR3 and stimulate CCR3+ cells. Eotaxin stimulates Th2 cells expressing CCR3. Other studies describe HIV-1 specific T cell cytotoxicity can be mediated by RANTES and Eotaxin through CCR3. CCR3 expressed on dendritic cells may have a biological role on cell-cell interaction during antigen presentation. CCR3 has been clustered as CD193 in the HLDA VIIIth workshop.

564189 Rev. 3
Format Details
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
564189 Rev.3
Citations & References
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Development References (9)

  1. Agrawal L, Maxwell CR, Peters PJ, et al. Complexity in human immunodeficiency virus type 1 (HIV-1) co-receptor usage: roles of CCR3 and CCR5 in HIV-1 infection of monocyte-derived macrophages and brain microglia. J Gen Virol. 2009; 90(3):710-722. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  2. Daugherty BL, Siciliano SJ, DeMartino JA, Malkowitz L, Sirotina A, Springer MS. Cloning, expression, and characterization of the human eosinophil eotaxin receptor. J Exp Med. 1996; 83(5):2349-2354. (Biology). View Reference
  3. Ghorpade A, Xia MQ, Hyman BT, et al. Role of the beta-chemokine receptors CCR3 and CCR5 in human immunodeficiency virus type 1 infection of monocytes and microglia. J Virol. 1998; 72(4):3351-3361. (Biology). View Reference
  4. Hadida F, Vieillard V, Autran B, Clark-Lewis I, Baggiolini M, Debre P. HIV-specific T cell cytotoxicity mediated by RANTES via the chemokine receptor CCR3. J Exp Med. 1998; 188(3):609-614. (Biology). View Reference
  5. Heath H, Qin S, Rao P, et al. Chemokine receptor usage by human eosinophils. The importance of CCR3 demonstrated using an antagonistic monoclonal antibody. J Clin Invest. 1997; 99(2):178-184. (Immunogen: Flow cytometry). View Reference
  6. Liu SM, Xavier R, Good KL, et al. Immune cell transcriptome datasets reveal novel leukocyte subset-specific genes and genes associated with allergic processes. J Allergy Clin Immunol. 2006; 118(2):496-503. (Clone-specific). View Reference
  7. Sallusto F, Mackay CR, Lanzavecchia A. Selective expression of the eotaxin receptor CCR3 by human T helper 2 cells. Science. 1997; 277(5334):2005-2007. (Biology). View Reference
  8. Sato K, Kawasaki H, Nagayama H, et al. CC chemokine receptors, CCR-1 and CCR-3, are potentially involved in antigen-presenting cell function of human peripheral blood monocyte-derived dendritic cells. Blood. 1999; 93(1):34-42. (Biology). View Reference
  9. Zimmermann N, Daugherty BL, Stark JM, Rothenberg ME. Molecular analysis of CCR-3 events in eosinophilic cells. J Immunol. 2000; 164(2):1055-1064. (Biology). View Reference
View All (9) View Less
564189 Rev. 3

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