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PE Mouse anti-Human PI-16
PE 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 the BD Pharmingen™ Human Regulatory T Cell Cocktail (Cat. No. 560249) containing FITC Mouse Anti-Human CD4, PE-Cy™7 Mouse Anti-Human CD25, and Alexa Fluor® 647 Mouse Anti-Human CD127 antibodies, and either PE Mouse IgG1, κ Isotype Control (Cat. No. 349043; Middle Panel) or PE Mouse Anti-Human PI-16 antibody (Cat. No. 563520; Right Panel). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots show the correlated expression patterns of PI-16 (or Ig Isotype control staining) versus CD25 for 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.

Multicolor flow cytometric analysis of PI-16 expression on human peripheral blood CD4+CD25+CD127low T cells. Whole human peripheral blood was stained with the BD Pharmingen™ Human Regulatory T Cell Cocktail (Cat. No. 560249) containing FITC Mouse Anti-Human CD4, PE-Cy™7 Mouse Anti-Human CD25, and Alexa Fluor® 647 Mouse Anti-Human CD127 antibodies, and either PE Mouse IgG1, κ Isotype Control (Cat. No. 349043; Middle Panel) or PE Mouse Anti-Human PI-16 antibody (Cat. No. 563520; Right Panel). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric contour plots show the correlated expression patterns of PI-16 (or Ig Isotype control staining) versus CD25 for 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_2738252
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 R-PE under optimum conditions, and unconjugated antibody and free PE 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. 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. Cy is a trademark of Amersham Biosciences Limited.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563520 Rev. 3
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.

563520 Rev. 3
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
563520 Rev.3
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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563520 Rev. 3

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.