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Mouse LRRC32/GARP Antibody

Catalog #: AF6229
3 Citations Datasheet / COA / SDS
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AF6229
AF6229-SP
Detection of Mouse LRRC32/GARP by Western Blot.
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Citations (3)
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Mouse LRRC32/GARP Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
See below
Immunohistochemistry
0.1-15 µg/mL
Immersion fixed paraffin-embedded sections of Mouse Spleen
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.

Scientific Data

Western Blot Detection of Mouse LRRC32/GARP antibody by Western Blot. View Larger

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 antibody in bEnd by Immunocytochemistry (ICC).3 Mouse Cell Line by Immunocytochemistry (ICC). View Larger

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.

Immunohistochemistry View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
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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
  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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Citations for Mouse LRRC32/GARP Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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  1. Thrombin contributes to cancer immune evasion via proteolysis of platelet-bound GARP to activate LTGF-beta
    Authors: Alessandra Metelli, Bill X. Wu, Brian Riesenberg, Silvia Guglietta, John D. Huck, Catherine Mills et al.
    Science Translational Medicine
  2. B lymphocytes confer immune tolerance via cell surface GARP-TGF-? complex
    Authors: CH Wallace, BX Wu, M Salem, EA Ansa-Addo, A Metelli, S Sun, G Gilkeson, MJ Shlomchik, B Liu, Z Li
    JCI Insight, 2018-04-05;3(7):.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Surface Expression of TGF-beta Docking Receptor GARP Promotes Oncogenesis and Immune Tolerance in Breast Cancer
    Authors: Alessandra Metelli, Bill X. Wu, Caroline W. Fugle, Saleh Rachidi, Shaoli Sun, Yongliang Zhang et al.
    Cancer Research

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