Detects mouse LRRC32/GARP in direct ELISAs. In direct ELISAs, approximately 20% cross-reactivity with recombinant human (rh) LRRC32 is observed and less than 5% cross-reactivity with rhLRRC3, rhLRRC4, and rhLRRC4B is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Mouse LRRC32/GARP Antibody

Detection of Mouse LRRC32/GARP by Western Blot.

Western blot shows lysates of mouse spleen non-B cells. PVDF Membrane was probed with 0.1 µg/mL of Sheep Anti-Mouse LRRC32/GARP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6229) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for LRRC32/GARP at approximately 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

LRRC32/GARP in bEnd.3 Mouse Cell Line.

LRRC32/GARP was detected in immersion fixed bEnd.3 mouse endothelioma cell line using Sheep Anti-Mouse LRRC32/ GARP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6229) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of LRRC32/GARP in Mouse Spleen.

LRRC32/GARP was detected in immersion fixed paraffin-embedded sections of Mouse Spleen using Sheep Anti-Mouse LRRC32/GARP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6229) at 0.3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using the Donkey Anti-Sheep IgG HRP Affinity Purified PAb (brown; Catalog # HAF016) and counterstained with hematoxylin (blue). Specific staining was localized to extracellular space. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Mouse LRRC32/GARP Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed bEnd.3 mouse endothelioma cell line

Immunohistochemistry

0.1-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Mouse Spleen

Western Blot

0.1 µg/mL
Sample: Mouse spleen non-B cells

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: LRRC32/GARP

Leucine-rich repeat protein 32 (LRRC32), also known as GARP (glycoprotein A repetitions predominant), is an 80 kDa type I transmembrane glycoprotein (1). Mature mouse LRRC32 consists of a 608 amino acid (aa) extracellular domain (ECD) that contains 22 leucine-rich repeats, a 21 aa transmembrane segment, and a 14 aa cytoplasmic domain (2-4). Within the ECD, mouse LRRC32 shares 80 and 94% aa sequence identity with human and rat LRRC32, respectively. LRRC32 is widely expressed during embryogenesis and on adult platelets (4, 5). Among T cells, it is selectively expressed on activated FOXP3+ regulatory T cells (Treg) (6-10). LRRC32 expression promotes the acquisition of a Treg phenotype including reduced cellular proliferation, reduced cytokine secretion, and the capacity to suppress the proliferation of naïve T cells (6-8). LRRC32 binds directly to the TGF-beta latency associated peptide (LAP) and tethers latent TGF-beta on the surface of activated Treg cells (9, 10). The presentation of TGF-beta on Tregs contributes to their ability to suppress naïve T cell proliferation (11).

References

Battaglia, M. and M.G. Roncarolo (2009) Eur. J. Immunol. 39:3296.
Ollendorff, V. et al. (1994) Cell Growth Differ. 5:213.
Bella, J. et al. (2008) Cell Mol Life Sci. 65:2307.
Roubin, R. et al. (1996) Int. J. Dev. Biol. 40:545.
Macaulay, I.C. et al. (2007) Blood 109:3260.
Wang, R. et al. (2008) PloS ONE 3:e2705.
Wang, R. et al. (2009) Proc. Natl. Acad. Sci. 106:13439.
Probst-Kepper, M. et al. (2009) J. Cell. Mol. Med. 13:3343.
Tran, D.Q. et al. (2009) Proc. Natl. Acad. Sci. 106:13445.
Stockis, J. et al. (2009) Eur. J. Immunol. 39:3315.
Vignali, D.A. et al. (2008) Nat. Rev. Immunol. 8:523.

Long Name

Leucine-rich Repeat Containing 32/Glycoprotein A Repetitions Predominant

Alternate Names

D11S833E, GARP, Garpin

Entrez Gene IDs

2615 (Human); 434215 (Mouse)

Gene Symbol

LRRC32

UniProt

NP_001106850

Additional LRRC32/GARP Products

Product Documents for Mouse LRRC32/GARP Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Mouse LRRC32/GARP Antibody

For research use only

Citations
Reviews

Mouse LRRC32/GARP Antibody

R&D Systems, part of Bio-Techne | Catalog # AF6229

Key Product Details

Species Reactivity

Validated:

Cited:

Applications