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APC Mouse anti-Human GARP
APC 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 APC Mouse IgG2b κ Isotype Control (mouse IgG2b, Cat. No. 555745) or APC Mouse anti-Human GARP (Cat. No. 562341). The cells were then fixed, permeabilized and stained with BD Horizon™ V450 Mouse Anti-Human FoxP3 (Cat. No. 560459) as per the recommended protocol. Two-color flow cytometric dot plots showing the correlated expression patterns of FoxP3 versus the Ig Isotype Control (Left Panel) or GARP (Right Panel) were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

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 APC Mouse IgG2b κ Isotype Control (mouse IgG2b, Cat. No. 555745) or APC Mouse anti-Human GARP (Cat. No. 562341). The cells were then fixed, permeabilized and stained with BD Horizon™ V450 Mouse Anti-Human FoxP3 (Cat. No. 560459) as per the recommended protocol. Two-color flow cytometric dot plots showing the correlated expression patterns of FoxP3 versus the Ig Isotype Control (Left Panel) or GARP (Right Panel) were derived from gated events with the forward and side light-scatter characteristics of intact 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_10926211
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 APC under optimum conditions, and unconjugated antibody and free APC were removed.

Recommended Assay Procedures

Suggested Staining Procedure for APC anti-Human GARP Antibody:

       1.  Harvest PBMC after stimulation (12 hours) with 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 X 10^6 cells either with the APC anti-Human GARP antibody (Cat. No. 562341) or with APC Mouse IgG2b, κ Isotype Control

            (Cat. No. 555745) for 30 minutes on ice, protected from light.

       4.  Wash cells twice with stain buffer.

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

            FoxP3 staining protocol.

        In brief,

             a.  Add 2 ml of 1 X 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 Anti-Human FoxP3 antibody (Cat. No. 560459) for 30-45 minutes.

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

        Note: BD Pharmingen™ Human FoxP3 Buffer Set (Cat. No. 560098) is a suggested companion product.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. 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).
562341 Rev. 1
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.

562341 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
562341 Rev.1
Citations & References
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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
562341 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.