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BV421 Rat Anti-Human CD294 (CRTH2)
BV421 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™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or a BD Horizon™ BV421 Rat Anti-Human CD294 antibody (Cat. No. 562992; Right 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 LSRFortessa™ Cell Analyzer 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™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Panel) or a BD Horizon™ BV421 Rat Anti-Human CD294 antibody (Cat. No. 562992; Right 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 LSRFortessa™ Cell Analyzer 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)
5 µl
VIII 80349
AB_2737937
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
562992 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 was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562992 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562992 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. (Clone-specific: Flow cytometry). 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. (Clone-specific: Flow cytometry). 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. (Immunogen: Blocking, Cell separation, Flow cytometry, Western blot). View Reference
562992 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.