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V450 Rat Anti-Human CD294 (CRTH2)
V450 Rat Anti-Human CD294 (CRTH2)

Multicolor 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 BD Horizon™ V450 Rat Anti-Human CD294 antibody (Cat. No. 561661 Right Panel) or with a BD Horizon™ V450 Rat IgG2a, κ Isotype Control (Cat. No. 560377; 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.

Multicolor 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 BD Horizon™ V450 Rat Anti-Human CD294 antibody (Cat. No. 561661 Right Panel) or with a BD Horizon™ V450 Rat IgG2a, κ Isotype Control (Cat. No. 560377; 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 Horizon™
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_10893005
Aqueous buffered solution containing protein stabilizer 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 BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
561661 Rev. 1
Antibody Details
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BM16

The BM16 monoclonal antibody specifically binds to CD294. CD294 is encoded by PTGDR2 (Prostaglandin D2 receptor 2) and is also known as CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2 cells), GPR44 (G protein-coupled receptor 44), DL1R, and DP2. CD294 is a member of the G protein-coupled leukocyte chemoattractant receptor family. CD294 is expressed on Th2 cells and type-2 innate lymphoid cells (ILC2), but not Th1 type cells. CD294 is detectable on CD4+ T cells in fresh PBMC but not on B cells and NK cells. CD294 is also expressed on peripheral blood basophils and eosinophils, suggesting its involvement allergic reactions. Phenotypic analysis of CD4+ T cells expressing CRTH2 demonstrated that they were also CD45RA-negative and CD45RO+ and CD25+. These cells produce Th2- but little or no Th1-type cytokines upon stimulation with PMA and Ionomycin.

The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

561661 Rev. 1
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V450
Violet 405 nm
405 nm
450 nm
561661 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
561661 Rev. 1

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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.