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BD Horizon™ BV421 Mouse Anti-Human GARP
Clone 7B11 (also known as CMSSC-7B11) (RUO)
Multicolor flow cytometric analysis of GARP expressed on activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were activated (3 days) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725) antibodies. The activated PBMC were washed and stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707) and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Left Panel) or BD Horizon™ BV421 Mouse Anti-Human GARP antibody (Cat. No. 563956/565969, Right Panel). The cells were then washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with PE Mouse Anti-Human FoxP3 antibody (Cat. No. 560046/560082). Two-color flow cytometric dot plots showing the correlated expression of FoxP3 versus GARP (or Ig Isotype control staining) were derived from CD4-positive events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis 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 were activated (3 days) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725) antibodies. The activated PBMC were washed and stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707) and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Left Panel) or BD Horizon™ BV421 Mouse Anti-Human GARP antibody (Cat. No. 563956/565969, Right Panel). The cells were then washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with PE Mouse Anti-Human FoxP3 antibody (Cat. No. 560046/560082). Two-color flow cytometric dot plots showing the correlated expression of FoxP3 versus GARP (or Ig Isotype control staining) were derived from CD4-positive events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis 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 were activated (3 days) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725) antibodies. The activated PBMC were washed and stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707) and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Left Panel) or BD Horizon™ BV421 Mouse Anti-Human GARP antibody (Cat. No. 563956/565969, Right Panel). The cells were then washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with PE Mouse Anti-Human FoxP3 antibody (Cat. No. 560046/560082). Two-color flow cytometric dot plots showing the correlated expression of FoxP3 versus GARP (or Ig Isotype control staining) were derived from CD4-positive events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis 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 were activated (3 days) with plate-bound Purified NA/LE Mouse Anti-Human CD3 (Cat. No. 555329) and soluble Purified NA/LE Mouse Anti-Human CD28 (Cat. No. 555725) antibodies. The activated PBMC were washed and stained with Alexa Fluor® 647 Mouse Anti-Human CD4 antibody (Cat. No. 557707) and either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748, Left Panel) or BD Horizon™ BV421 Mouse Anti-Human GARP antibody (Cat. No. 563956/565969, Right Panel). The cells were then washed, fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with PE Mouse Anti-Human FoxP3 antibody (Cat. No. 560046/560082). Two-color flow cytometric dot plots showing the correlated expression of FoxP3 versus GARP (or Ig Isotype control staining) were derived from CD4-positive events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
LRRC32; Garpin; Glycoprotein A repetitions predominant
Human (QC Testing)
Mouse IgG2b, κ
Recombinant Human LRRC32
Flow cytometry (Routinely Tested)
5 µl
AB_2738508
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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
BV421 Mouse IgG2b, κ Isotype Control RUO
Size
50 µg
Cat No.
562748
Purified NA/LE Mouse Anti-Human CD3 RUO
Size
0.5 mg
Cat No.
555329
Purified NA/LE Mouse Anti-Human CD28 RUO
Size
0.5 mg
Cat No.
555725
563956 Rev. 2
Antibody Details
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.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.
563956 Rev. 2
Format Details
BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BV421
407 nm
423 nm
563956 Rev.2
Citations & References
View product citations for antibody "563956" on CiteAb
Development References (2)
-
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
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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
563956 Rev. 2
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.