Mouse Lgr5/GPR49 Antibody

Published as Clone RD20.
  
  • Species Reactivity
    Mouse
  • Specificity
    Detects mouse Lgr5/GPR49 in ELISA. Stains mouse Lgr5 transfected cells but not irrelevant transfectants in Flow Cytometry and Immunocytochemistry. This antibody, also known as "RD20", has been found to detect an epitope in C-terminal LRR cap of LGR5 (Ref. 1).
  • Source
    Monoclonal Rat IgG2B Clone # 803420
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant mouse Lgr5/GPR49
    Lys375-Pro547
    Accession # Q9Z1P4
  • 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
    2.5 µg/106 cells
    See below
  • Agonist Activity
    Peng, W.C., et al. (2013) Cell Reports. 3:1885.
  • 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 Lgr5/GPR49 in HEK293 Human Cell Line Transfected with Mouse Lgr5/GPR49 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with mouse Lgr5/GPR49 and eGFP was stained with and either (A) Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) or (B) Rat IgG2B Isotype Control (Catalog # MAB0061) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113).
Immunocytochemistry
Lgr5/GPR49 in HEK293 Human Cell Line Transfected with Mouse Lgf5/GPR49. Lgr5/GPR49 was detected in immersion fixed HEK293 human embryonic kidney cell line transfected with GFP (green) tagged mouse LGR5 using Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunocytochemistry
Lgr5/GPR49 in Mouse Cells Transfected with Mouse Lgr5/GPR49. Lgr5/GPR49 was detected in immersion fixed MYC tagged mouse transfectants using Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240). Cells were stained using an anti-Mouse HRP-conjugated secondary antibody (brown). Image courtesy of Dr. Hans Clevers, Hubrecht Institute, The Netherlands.
Mouse Lgr5/GPR49 Antibody Induces Activity. Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8240) induces TOPflash activity in the HEK293 human embryonic kidney cell line stably expressing LGR5 in the presence of Wnt-3a, but in the absence of R-Spondins. (Data courtesy of  Dr. Wim de Lau and Dr. Hans Clevers, Hubrecht Institute, The Netherlands. See Reference 1.)
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: Lgr5/GPR49
Leucine‑rich repeat G‑protein‑coupled receptor 5 (Lgr5), also called GPR49, is a 907 amino acid (aa), approximately 97 kDa (calculated), seven‑transmembrane glycoprotein receptor in the Lgr family of cell surface receptors. The subfamily of Lgrs comprising Lgr4, Lgr5, and Lgr6 are G‑protein‑independent mediators of the potentiating effect of R‑Spondins on Wnt signaling (2). Lgr5 binds and forms complexes with R‑Spondins, Frizzled Wnt receptors and LRP Wnt co‑receptors. The region of the mouse Lgr5 long extracellular domain used as an immunogen shares 90% and 95% amino acid sequence identity with human and rat Lgr5, respectively. Lgr5 is found on embryonic and adult epithelial stem cells (3). Lgr5+ stem cells can produce all epithelial cell types of the intestinal crypts (4). It is upregulated in stem cells that give rise to cancers such as intestinal, hepatocellular, pancreatic and ovarian carcinomas (5,6). This antibody has been referred to as "RD20" by Peng et al.(1).
  • References:
    1. Peng et al. (2013) Cell Rep. 3(6):1885.
    2. de Lau et al. (2014) Genes Dev. 28(4):305.
    3. Barker et al. (2013) Development 140(12):2484.
    4. Clevers (2013) Nature 495(7439):53.
    5. Wu et al. (2014) Nat Commun 5:3149.
    6. Jang et al. (2013) PLoS One 8(12):e82390495.
  • Long Name:
    Leucine-rich Repeat Containing G Protein-coupled Receptor 5
  • Entrez Gene IDs:
    8549 (Human); 14160 (Mouse); 299802 (Rat)
  • Alternate Names:
    FEX; G protein-coupled receptor 49; GPR49; GPR49G-protein coupled receptor HG38; GPR67; G-protein coupled receptor 49; G-protein coupled receptor 67; GRP49; HG38; leucine-rich repeat containing G protein-coupled receptor 5; leucine-rich repeat-containing G protein-coupled receptor 5; leucine-rich repeat-containing G-protein coupled receptor 5; Lgr5; MGC117008; orphan G protein-coupled receptor HG38
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