Human Lgr4/GPR48 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Lgr4/GPR48 in Lgr4-transfected cell-based ELISA. Stains human Lgr4/GPR48 transfected but not irrelevant transfected cells in flow cytometry.
  • Source
    Monoclonal Mouse IgG2B Clone # 852229
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    NS0 mouse myeloma cell line transfected with human Lgr4/GPR48
    Met1-Asp951
    Accession # Q9BXB1
  • 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
  • Flow Cytometry
    0.25 µg/106 cells
    See below
  • Blockade of Receptor-ligand Interaction
    In a functional flow cytometry test, 2.5 μg/mL of Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody completely blocks binding of biotinylated Recombinant Human R-Spondin 4 (100 ng/mL) to HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48.
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 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 Lgr4/GPR48 in HEK293 Human Cell Line Transfected with Human Lgr4/GPR48 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48 and eGFP was stained with and either
(A) Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750) or (B) Mouse IgG2B Flow Cytometry Isotype Control (Catalog # MAB0041) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody(Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.
Immunocytochemistry
Lgr4/GPR48 in HT‑29 Human Cell Line. Lgr4/GPR48 was detected in immersion fixed HT‑29 human colon adenocarcinoma cell line using Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Blockade of Receptor-ligand Interaction
R-Spondin 4 Binding to Lgr4/GPR48-transfected HEK293 Human Cell Line is Blocked by Human Lgr4/GPR48 Antibody. In a functional flow cytometry test, biotinylated Recombinant Human R-Spondin 4 (100 ng/mL) binds to HEK293 human embryonic kidney cell line transfected with human Lgr4/GPR48 (dark orange histogram). Binding is completely blocked (light orange histogram) by 2.5 μg/mL of Mouse Anti-Human Lgr4/GPR48 Monoclonal Antibody (Catalog # MAB7750). Mouse IgG2B Isotype Control (Catalog # MAB004) at 2.5 μg/mL was used as a control (blue line). Cells were stained with Streptavidin-APC (Catalog # F0050).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • 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: Lgr4/GPR48

Lgr4 (Leucine-rich repeat GPR 4), also known as GPR48 (G-Protein-coupled Receptor 48), is a seven-transmembrane glycoprotein receptor in the Lgr family of cell surface receptors (1, 2). While this family includes receptors for hormones such as LH, FSH, TSH, and HCG, the subfamily comprising Lgr4, Lgr5, and Lgr6 are G-protein-independent mediators of the potentiating effect of R-Spondins on Wnt signaling (1-6). Lgr4 binds and forms complexes with R-Spondins, Frizzled Wnt receptors and LRP Wnt co-receptors (5). It acts at least in part by enhancing Wnt-dependent LRP phosphorylation, internalization of LRPs, and accumulation of beta -catenin (3, 4). Human Lgr4 cDNA encodes 951 amino acids (aa), including a long N-terminal Extracellular Domain (ECD, aa 25-544) with 16-17 LRR domains that mediate ligand interaction (1). The LRR-containing ECD of human Lgr4 shares 93% aa sequence identity with mouse, rat and bovine Lgr4, and 50-60% aa identity with human Lgr5 and Lgr6. Lgr4 is widely expressed in both embryo and adult. Expression of Lgr4 mRNA in adult humans is highest in pancreas, followed by liver, heart, muscle, brain, and placenta (1). In rodents, embryonic and adult expression includes liver, kidney, adrenals, bone/cartilage, and heart (2, 7-9). Lgr4 deletion in the mouse affects development in areas of expression, for example, inhibiting fetal liver definitive erythropoiesis (9). Deletion of Lgr4 specifically from stem and progenitor cells in intestinal crypts induces loss of crypts due to insufficient Wnt signaling (5, 6). Lgr4 may be over-expressed in carcinomas and may promote invasiveness and metastasis by down-regulating p27Kip1 expression (10).

  • References:
    1. Loh, E.D. et al. (2001) Biochem. Biophys. Res. Commun. 282:757.
    2. Hsu, S.Y. et al. (1998) Mol. Endocrinol. 12:1830.
    3. Carmon, K.S. et al. (2011) Proc. Natl. Acad. Sci. USA 108:11452.
    4. Glinka, A. et al. (2011) EMBO Rep. 12:1055.
    5. de Lau, W. et al. (2011) Nature 476:293.
    6. Ruffner, H. et al. (2012) PLoS ONE 7:e40975.
    7. Van Schoore, G. et al. (2005) Histochem. Cell Biol. 124:35.
    8. Mazerbourg, S. et al. (2004) Mol. Endocrinol. 18:2241.
    9. Song, H. et al. (2008) J. Biol. Chem. 283:36687.
    10. Gao, Y. et al. (2006) Cancer Res. 66:11623.
  • Long Name:
    Leucine-rich Repeat Containing G Protein-coupled Receptor 4
  • Entrez Gene IDs:
    55366 (Human); 107515 (Mouse); 286994 (Rat)
  • Alternate Names:
    GPR48; GPR48G protein-coupled receptor 48; G-protein coupled receptor 48; leucine-rich repeat containing G protein-coupled receptor 4; Lgr4
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