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      Alexa Fluor® 647 Mouse Anti-Human PI-16
      Alexa Fluor® 647 Mouse Anti-Human PI-16
      Multicolor flow cytometric analysis of PI-16 expression on human peripheral blood CD4+CD25+CD127low T cells. Whole human peripheral blood was stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842), BD Horizon™ BV421 Mouse Anti-Human CD25 (Cat. No. 564033), and PE Mouse Anti-Human CD127 (552543/557938/561028) antibodies, and either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; Middle Panel) or Alexa Fluor® 647 Mouse Anti-Human PI-16 antibody (Cat. No. 564969; Right Panel). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of PI-16 (or Ig Isotype control staining) versus CD25 were derived from CD4+CD25+CD127low-gated events (Left Panel) with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Alexa Fluor® 647 Mouse Anti-Human PI-16
      Multicolor flow cytometric analysis of PI-16 expression on human peripheral blood CD4+CD25+CD127low T cells. Whole human peripheral blood was stained with FITC Mouse Anti-Human CD4 (Cat. No. 555346/561005/561842), BD Horizon™ BV421 Mouse Anti-Human CD25 (Cat. No. 564033), and PE Mouse Anti-Human CD127 (552543/557938/561028) antibodies, and either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; Middle Panel) or Alexa Fluor® 647 Mouse Anti-Human PI-16 antibody (Cat. No. 564969; Right Panel). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots showing the correlated expression of PI-16 (or Ig Isotype control staining) versus CD25 were derived from CD4+CD25+CD127low-gated events (Left Panel) with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Pharmingen™
      CD364; PI16; Peptidase inhibitor 16; CRISP-9; PSPBP; PSP94-binding protein
      Human (QC Testing)
      Mouse IgG1, κ
      Human PI-16 Transfected Cell Line
      Flow cytometry (Routinely Tested)
      5 µl
      AB_2739028
      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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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      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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564969 Rev. 1
      Antibody Details
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      CRCBT-02-001

      The CRCBT-02-001 monoclonal antibody specifically binds to human CD364 which is also known as Peptidase inhibitor 16 (PI-16) or Protease inhibitor 16. PI-16 is a glycophosphatidylinositol-linked, 436 amino acid surface glycoprotein. It is a member of the cysteine-rich secretory protein (CRISP) family and is likewise known as CRISP9. PI-16 is a putative serine protease inhibitor and can serve as a binding protein for PSP94 (prostate secretory protein of 94 amino acids) termed PSPBP. Serum levels of PSP94 and PI-16 can reportedly serve as key targets for prostate cancer research. PI-16 is highly expressed by the activated/memory, FoxP3-bright subsets of regulatory T cells. PI-16-positive regulatory T cells expressed uniformly high levels of the chemokine receptors, CCR4 and CCR6, when compared with PI-16-negative T regulatory cells. The results suggest that the CCR4+CCR6+ regulatory T cells are capable of homing to inflammatory sites and regulating effector T cells such as CCR4+CCR6+ Th17-like cells. PI-16  is also expressed by subsets of memory CD4+ and CD8-bright  T cells. It is not expressed by B cells or monocytes.

                      

      564969 Rev. 1
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-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.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      564969 Rev.1
      Citations & References
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      Development References (5)

      1. Grose RH, Millard DJ, Mavrangelos C, et al. Comparison of blood and synovial fluid th17 and novel peptidase inhibitor 16 Treg cell subsets in juvenile idiopathic arthritis. J Rheumatol. 2012; 39(10):2021-2031. (Clone-specific: Flow cytometry). View Reference
      2. Nicholson IC, Mavrangelos C, Bird DR, et al. PI16 is expressed by a subset of human memory Treg with enhanced migration to CCL17 and CCL20. Cell Immunol. 2012; 275(1-2):12-18. (Immunogen: Flow cytometry). View Reference
      3. Reeves JR, Dulude H, Panchal C, Daigneault L, Ramnani DM. Prognostic value of prostate secretory protein of 94 amino acids and its binding protein after radical prostatectomy. Clin Cancer Res. 2006; 12(20 Pt 1):6018-6022. (Clone-specific). View Reference
      4. Sadlon TJ, Wilkinson BG, Pederson S, et al. Genome-wide identification of human FOXP3 target genes in natural regulatory T cells. J Immunol. 2010; 185(2):1071-1081. (Biology). View Reference
      5. Whitaker HC, Warren AY, Eeles R, Kote-Jarai Z, Neal DE. The potential value of microseminoprotein-beta as a prostate cancer biomarker and therapeutic target. Prostate. 2012; 70(3):333-340. (Biology). View Reference
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      564969 Rev. 1

      Please refer to Support Documents for Quality Certificates


      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.

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