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      Purified Rat anti-Human Lgr5 (CRL Region)

      BD Pharmingen™ Purified Rat anti-Human Lgr5 (CRL Region)

      Clone 1D9A5 (also known as 1D9) (RUO)

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      Purified Rat anti-Human Lgr5 (CRL Region)
      Western blot analysis of human Lgr5. Lysate from colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) was probed with Purified Rat anti-Human Lgr5 (CRL Region) monoclonal antibody (Cat. No. 562733) at titrations of 1.0 (lane 1), 0.5 (lane 2), and 0.25 μg/ml (lane 3). Lgr5 is identified as a strong band of ~110 kDa. The protein was detected using HRP Goat Anti-Rat Ig (Cat. No. 554017) and a chemiluminescent detection system.
      Purified Rat anti-Human Lgr5 (CRL Region)
      Immunofluorescent staining of human Lgr5. Colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) were stained with Purified Rat anti-Human Lgr5 (CRL Region) monoclonal antibody (Cat. No. 562733) (pseudo colored green) at 5 ug/mL. The second-step reagent was Alexa Fluor® 488 goat anti-Rat IgG (Life Technologies). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
      Purified Rat anti-Human Lgr5 (CRL Region) Purified Rat anti-Human Lgr5 (CRL Region)
      Western blot analysis of human Lgr5. Lysate from colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) was probed with Purified Rat anti-Human Lgr5 (CRL Region) monoclonal antibody (Cat. No. 562733) at titrations of 1.0 (lane 1), 0.5 (lane 2), and 0.25 μg/ml (lane 3). Lgr5 is identified as a strong band of ~110 kDa. The protein was detected using HRP Goat Anti-Rat Ig (Cat. No. 554017) and a chemiluminescent detection system.
      Immunofluorescent staining of human Lgr5. Colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) were stained with Purified Rat anti-Human Lgr5 (CRL Region) monoclonal antibody (Cat. No. 562733) (pseudo colored green) at 5 ug/mL. The second-step reagent was Alexa Fluor® 488 goat anti-Rat IgG (Life Technologies). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
      Product Details
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      BD Pharmingen™
      GPR49, GPR67, HG38
      Human (QC Testing), Mouse (Lack of Reactivity Confirmed in Development)
      Rat IgG2b, λ
      Human LGR5 DNA
      Western blot (Routinely Tested), Bioimaging, Flow cytometry, Immunofluorescence (Tested During Development), Immunohistochemistry-formalin (antigen retrieval required), Immunohistochemistry-frozen (Not Recommended)
      ~110 kDa
      0.5 mg/ml
      AB_2737754
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
      4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
      5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
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      562733 Rev. 1
      Antibody Details
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      1D9A5

      Lgr5 (leucine-rich-repeat-containing G-protein-coupled receptor 5) is a seven transmembrane-domain receptor that is a target gene for Wnt and marks stem cells in the small intestine and colon, stomach, and hair follicle. Lgr5 was initially identified as a potential stem cell marker due to restricted expression of Lgr5 in the intestinal crypt and labeling of rapidly cycling cells of the colon and intestine. Using both lineage tracing and organiod culture experiments, Lgr5 positive cells are capable of generating all types of the small intestine epithelium hence indicating that Lgr5 marks stem cells of the small intestine and colon. R-spondin growth factors, which are secreted agonists of the Wnt pathway, bind Lgr5. The binding of R-spondins to Lgr5 leads to recruitment of the Frizzled/LRP Wnt receptor complex, which binds to Wnt ligands and leads to downstream Wnt signaling. Lgr5 is up-regulated in colon and ovarian cancers and has been implicated in promotion of tumor growth and metastasis.

      The 1D9A5 monoclonal antibody recognizes an epitope in the Cysteine-rich linker (CRL) region of Human Lgr5.

      562733 Rev. 1
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      562733 Rev.1
      Citations & References
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      View product citations for antibody "562733" on CiteAb

      Development References (8)

      1. Barker N, Huch M, Kujala P, et al. Lgr5(+ve) stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell. 2010; 6(1):25-36. (Biology). View Reference
      2. Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature. 2007; 449(7165):1003-1007. (Biology). View Reference
      3. Carmon KS, Gong X, Lin Q, Thomas A, Liu Q. R-spondins function as ligands of the orphan receptors LGR4 and LGR5 to regulate Wnt/{beta}-catenin signaling. Proc Natl Acad Sci U S A. 2011; 108(28):11452-11457. (Biology). View Reference
      4. Jaks V, Barker N, Kasper M, et al. Lgr5 marks cycling, yet long-lived, hair follicle stem cells. Nat Genet. 2008; 40(11):1291-1299. (Biology). View Reference
      5. Kemper K, Prasetyanti PR, de Lau W, Rodermond H, Clevers H, Medema JP. Monoclonal antibodies against Lgr5 identify human colorectal cancer stem cells. Stem Cells. 2012; 30(11):2378-2386. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
      6. Merlos-Suárez A, Barriga FM, Jung P et al. The intestinal stem cell signature identifies colorectal cancer stem cells and predicts disease relapse. Cell Stem Cell. 2011; 8(5):511-524. (Biology). View Reference
      7. Yui S, Nakamura T, Sato T, et al. Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5(+) stem cell. Nat Med. 2012; 18(4):618-623. (Biology). View Reference
      8. de Lau W, Barker N, Low TY, et al. Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling. Nature. 2011; 476(7360):293-297. (Clone-specific). View Reference
      View All (8) View Less
      562733 Rev. 1

      Please refer to Support Documents for Quality Certificates

       

      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

       

      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.