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APC Rat Anti-Human CD294 (CRTH2)
APC Rat Anti-Human CD294 (CRTH2)
Multiparameter flow cytometric analysis of CD294 (CRTH2) expression on human peripheral blood leucocyte populations. Whole blood was stained with either APC Rat IgG2a κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Human CD294 (CRTH2) antibody (Cat. No. 566785; Right Plot) at 1 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A pseudocolor density dot plot showing the correlated expression of CD294 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD294 (CRTH2) expression on human peripheral blood leucocyte populations. Whole blood was stained with either APC Rat IgG2a κ Isotype Control (Cat. No. 553932; Left Plot) or APC Rat Anti-Human CD294 (CRTH2) antibody (Cat. No. 566785; Right Plot) at 1 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A pseudocolor density dot plot showing the correlated expression of CD294 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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_2869865
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 to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566785 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.

        

566785 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
566785 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
566785 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.