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      PE Mouse anti-Human GARP
      PE Mouse anti-Human GARP
      Multicolor flow cytometric analysis of GARP expressed on activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were activated (12 hours) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725). The activated PBMC were washed and stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 556656) or PE Mouse Anti-Human GARP (Cat. No. 562150). The cells were then washed and stained with BD Horizon™ V450 Mouse Anti-Human FoxP3 (Cat. No. 560459) using FoxP3 staining protocol. Two-color flow cytometric dot plots showing the correlated expression staining patterns of V450 FoxP3 versus PE anti-Human GARP (right panel) or PE Mouse IgG2b, κ isotype control (left panel) were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      PE Mouse anti-Human GARP
      Multicolor flow cytometric analysis of GARP expressed on activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were activated (12 hours) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725). The activated PBMC were washed and stained with either PE Mouse IgG2b, κ Isotype Control (Cat. No. 556656) or PE Mouse Anti-Human GARP (Cat. No. 562150). The cells were then washed and stained with BD Horizon™ V450 Mouse Anti-Human FoxP3 (Cat. No. 560459) using FoxP3 staining protocol. Two-color flow cytometric dot plots showing the correlated expression staining patterns of V450 FoxP3 versus PE anti-Human GARP (right panel) or PE Mouse IgG2b, κ isotype control (left panel) were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Pharmingen™
      LRRC32; Garpin; Glycoprotein A repetitions predominant
      Human (QC Testing)
      Mouse IgG2b, κ
      Recombinant Human LRRC32
      Flow cytometry (Routinely Tested)
      5 µl
      2615
      AB_10893997
      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.
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      Recommended Assay Procedures

      Suggested Staining Procedure for PE Mouse anti-Human GARP Antibody:

             1.  Harvest PBMC after stimulation (12 hours) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and Purified

                  NA/LE Mouse Anti-Human CD28 (Cat. No. 555725).

             2.  Wash the cells twice with stain buffer (eg. BD Pharmingen™ Stain Buffer (FBS), Cat. No. 554656).

             3.  Stain 1 × 10^6 cells either with the PE Mouse anti-Human GARP antibody (Cat. No. 562150) or with PE Mouse IgG2b, κ Isotype control

                  (Cat. No. 556656) for 30 minutes on ice, protected from light.

             4.  Wash cells twice with stain buffer.

             5.  Stain with BD Horizon™ V450 Mouse anti-Human FoxP3. Refer to the Technical Data Sheet of Cat. No. 560459 for detailed FoxP3

                  staining protocol.

                  In brief,

                   a.  Add 2 ml of 1× FoxP3 buffer A to the cell pellet.

                   b.  Centrifuge and incubate in 0.5 ml of buffer C for 30 minutes.

                   c.  Wash twice with stain buffer and stain with V450 FoxP3 antibody (Cat. No. 560459) for 30-45 minutes.

                   d.  Wash twice with stain buffer and acquire on the Flow cyotmeter.

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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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      562150 Rev. 3
      Antibody Details
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      7B11

      The 7B11 (also known as CMSSC-7B11) monoclonal antibody specifically binds to human GARP (Glycoprotein A repetitions predominant). The LRRC32 (Leucine rich repeat containing 32) gene encodes the 662 amino acid-residue, 80 kDa transmembrane GARP glycoprotein that has an extracellular region composed primarily of 20 leucine-rich repeats. GARP is specifically expressed on Treg cells activated through the T cell receptor (TCR). Ectopic expression of GARP in human naïve T cells inhibited their proliferation and cytokine secretion upon TCR activation. Remarkably, GARP over-expression in naïve T cells induced expression of FoxP3 and endowed them with a partial suppressive function. The extracellular, but not the cytoplasmic region, of GARP was necessary for these functions. GARP serves as a receptor for latent TGF-beta which may play a role in the suppressive action of Treg cells. GARP is also expressed on platelets and other tissues, however the function on these cells is not known.

      562150 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.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      562150 Rev.3
      Citations & References
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      View product citations for antibody "562150" on CiteAb

      Development References (2)

      1. Ollendorff V, Noguchi T, deLapeyriere O, Birnbaum D. The GARP gene encodes a new member of the family of leucine-rich repeat-containing proteins. Cell Growth Differ. 1994; 5(2):213-219. (Biology). View Reference
      2. Stockis J, Colau D, Coulie PG, Lucas S. Membrane protein GARP is a receptor for latent TGF-beta on the surface of activated human Treg. Eur J Immunol. 2009; 39(12):3315-3322. (Biology). View Reference
      562150 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.