Mouse LRRC32/GARP Antibody

  
  • Species Reactivity
    Mouse
  • 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
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant mouse LRRC32/GARP
    Ile18-Asn628
    Accession # NP_001106850
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    Immersion fixed frozen sections of mouse thymus
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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.
Immunocytochemistry
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 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.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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:
    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.
  • Long Name:
    Leucine-rich Repeat Containing 32/Glycoprotein A Repetitions Predominant
  • Entrez Gene IDs:
    2615 (Human); 434215 (Mouse)
  • Alternate Names:
    D11S833E; D11S833Eleucine-rich repeat-containing protein 32; GARP; GARPGarpin; Garpin; Glycoprotein A repetitions predominantgarpin; leucine rich repeat containing 32; LRRC32
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