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PerCP-Cy™5.5 Rat Anti-Human CD294 (CRTH2)
PerCP-Cy™5.5 Rat Anti-Human CD294 (CRTH2)

Flow cytometric analysis of CD294 expression on human peripheral blood cells. Whole blood was stained with PE Mouse anti-Human CD45 (Cat. No. 555483/560975) and either PerCP-Cy™5.5 Rat Anti-Human CD294 antibody (Cat. No. 561660; Right Panel) or with a PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color dot plots showing the correlated expression of CD45 versus CD294 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.

Flow cytometric analysis of CD294 expression on human peripheral blood cells. Whole blood was stained with PE Mouse anti-Human CD45 (Cat. No. 555483/560975) and either PerCP-Cy™5.5 Rat Anti-Human CD294 antibody (Cat. No. 561660; Right Panel) or with a PerCP-Cy™5.5 Rat IgG2a, κ Isotype Control (Cat. No. 550765; Left Panel). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two-color dot plots showing the correlated expression of CD45 versus CD294 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometry System.

Product Details
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BD Pharmingen™
CRTH2; PTGDR2 ; Prostaglandin D2 receptor 2; DL1R; DP2; GPR44
Human (QC Testing)
Rat WI, also known as Wistar (outbred) IgG2a, κ
Human CRTH2 Transfected Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
VIII 80349
AB_10894394
Aqueous buffered solution containing ≤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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. 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.
  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. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  10. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
561660 Rev. 1
Antibody Details
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BM16

The BM16 monoclonal antibody specifically binds to CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells). CRTH2 is a member of the G protein-coupled leukocyte chemoattractant receptor family and it is expressed on Th2, but not Th1 lineage cells. It is detectable on CD4+ T cells in fresh PBMC and no significant expression on B cells and NK cells. CRTH2 is also expressed on peripheral blood basophils and eosinophils, suggesting its involvement allergic reactions. Phenotypic analysis of CD4+ T cells expressing CRTH2 demonstrated CD45RA-, CD45RO+ and CD25+. These cells produce Th2- but little or no Th1-type cytokines upon stimulation with PMA and Ionomycin.

561660 Rev. 1
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
561660 Rev.1
Citations & References
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Development References (3)

  1. Cosmi L, Annunziato F, Galli MIG , Maggi RME , Nagata K, Romagnani S. CRTH2 is the most reliable marker for the detection of circulating human type 2 Th and type 2 T cytotoxic cells in health and disease. Eur J Immunol. 2000; 30(10):2972-2979. (Biology). View Reference
  2. Nagata K, Hirai H, Tanaka K, et al. CRTH2, an orphan receptor of T-helper-2-cells, is expressed on basophils and eosinophils and responds to mast cell-derived factor(s). FEBS Lett. 1999; 459(2):195-199. (Biology). View Reference
  3. Nagata K, Tanaka K, Ogawa K, et al. Selective expression of a novel surface molecule by human Th2 cells in vivo. J Immunol. 1999; 162(3):1278-1286. (Biology). View Reference
561660 Rev. 1

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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.