LRRC32/GARP Antibody

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 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 Northern­Lights™ 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.

• Reactivity: Mu
• Applications: WB, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

LRRC32/GARP Antibody Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant mouse LRRC32/GARP
  Ile18-Asn628
  Accession # NP_001106850
• Specificity: 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: LRRC32
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Immunocytochemistry 5-15 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for LRRC32/GARP Antibody

• D11S833E
• D11S833Eleucine-rich repeat-containing protein 32
• GARP
• GARPGarpin
• Garpin
• Glycoprotein A repetitions predominantgarpin
• leucine rich repeat containing 32
• LRRC32

Background

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).

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for LRRC32/GARP Antibody (AF6229)(1)

Publication using AF6229

• CH Wallace, BX Wu, M Salem, EA Ansa-Addo, A Metelli, S Sun, G Gilkeson, MJ Shlomchik, B Liu, Z Li B lymphocytes confer immune tolerance via cell surface GARP-TGF-? complex JCI Insight, 2018;3(7):. 2018 [PMID: 29618665] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: LRRC32
• Uniprot:
  • NP_001106850()