Clone:
REA139
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
LRRC32, Garpin, Glycoprotein A repetitions predominant

Extended validation for GARP (LRRC32) Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA139
725226-
F011-5++
YGIC86++
Cells were incubated with an excess of purified unconjugated GARP (LRRC32) (REA139) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for Anti-GARP (LRRC32). Splenocytes from C57BL/6 mice stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with Anti-GARP (LRRC32) antibodies and with a suitable counterstaining. As a control, Anti-GARP (LRRC32) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Anti-GARP (LRRC32). Splenocytes from C57BL/6 mice stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with Anti-GARP (LRRC32) antibodies and with a suitable counterstaining. As a control, Anti-GARP (LRRC32) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Anti-GARP (LRRC32). Splenocytes from C57BL/6 mice stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with Anti-GARP (LRRC32) antibodies and with a suitable counterstaining. As a control, Anti-GARP (LRRC32) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Anti-GARP (LRRC32). Splenocytes from C57BL/6 mice stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with Anti-GARP (LRRC32) antibodies and with a suitable counterstaining. As a control, Anti-GARP (LRRC32) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for Anti-GARP (LRRC32). Splenocytes from C57BL/6 mice stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 24 hours were stained with Anti-GARP (LRRC32) antibodies and with a suitable counterstaining. As a control, Anti-GARP (LRRC32) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. CD4+ cells were pregated for the analysis. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using GARP (LRRC32) (REA139). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using GARP (LRRC32) (REA139). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using GARP (LRRC32) (REA139). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for GARP (LRRC32) Antibody, anti-mouse, REAfinity™

Overview

Clone REA139 recognizes murine GARP, a 80 KDa type I membrane glycoprotein which belongs to the family of leucine rich repeat proteins. Expression of GARP protein is found on megakaryocytes and platelets. Activated GARP has been found to be associated with latency associated peptide (LAP), the prodomain responsible for conferring latency to nonactive TGFβ. Thus, GARP as a latent TGFβ binding protein (LTBP) regulates the surface expression of TGFβ and serves as a carrier of LAP, at the cell surface, to immune response active sites, where TGFβ is eventually activated and released to exert further physiological functions such as differentiation of regulatory T cells.
Additional information: Clone REA139 displays negligible binding to Fc receptors.

Alternative names

LRRC32, Garpin, Glycoprotein A repetitions predominant

Detailed product information

Technical specifications

CloneREA139
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenGARP (LRRC32)
Alternative names of antigenLRRC32, Garpin, Glycoprotein A repetitions predominant
Molecular mass of antigen [kDa]72
Distribution of antigenT cells, megakaryocytes, platelets
Entrez Gene ID434215
RRIDAB_2651809, AB_2651810, AB_2651811, AB_2651812, AB_2801752

References for GARP (LRRC32) Antibody, anti-mouse, REAfinity™

Publications

  1. Tran, D. Q. et al. (2009)
    GARP (LRRC32) is essential for the surface expression of latent TGF-β on platelets and activated FOXP3
    +
    regulatory T cells.
    Proc. Natl. Acad. Sci. U.S.A. 106: 13445-13450
  2. Battaglia, M. et al. (2009) The Tregs' world according to GARP. Eur. J. Immunol. 39(12): 3296-3300
  3. Vermeersch, E. et al. (2017) The role of platelet and endothelial GARP in thrombosis and hemostasis. PLoS One 12(13): e0173329
  4. Wang, R. et al. (2008) Identification of a regulatory T cell specific cell surface molecule that mediates suppressive signals and induces Foxp3 expression. PLoS One 3(7): e2705
  5. Ollendorff, V. et al. (1994) The GARP gene encodes a new member of the family of leucine-rich repeat-containing proteins. Cell Growth Differ. 5: 213-219

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