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Biotin Rat anti-Human Lgr5 (Central LRR)
Biotin Rat anti-Human Lgr5 (Central LRR)

Flow cytometric analysis of LGR5 expression on human LGR5-transfected cells. LS 174T colorectal adenocarcinoma cells transfected with human LGR5 (cells from Dr. Hans Clevers, Hubrecht Institute, Left Panel) and parental LS 174T cells, which express low levels of endogenous LGR5 (ATCC CL-188, Right Panel), were harvested using Accutase™ Cell Detachment Solution (Cat. No. 561527). The cells were stained with either Biotin Rat IgG2b, κ Isotype Control (Cat. No. 553987; dashed line histogram) or Biotin Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562904; solid line histogram) at matched concentrations. The cells were then washed and stained with PE Streptavidin (Cat. No. 554061) as the secondary detection reagent. The fluorescence histograms were derived from gated events based on the light scattering characteristics of viable LS 174T cells. Flow cytometry was performed on a BD FACSCanto™ II Flow Cytometry System.

Flow cytometric analysis of LGR5 expression on human LGR5-transfected cells. LS 174T colorectal adenocarcinoma cells transfected with human LGR5 (cells from Dr. Hans Clevers, Hubrecht Institute, Left Panel) and parental LS 174T cells, which express low levels of endogenous LGR5 (ATCC CL-188, Right Panel), were harvested using Accutase™ Cell Detachment Solution (Cat. No. 561527). The cells were stained with either Biotin Rat IgG2b, κ Isotype Control (Cat. No. 553987; dashed line histogram) or Biotin Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562904; solid line histogram) at matched concentrations. The cells were then washed and stained with PE Streptavidin (Cat. No. 554061) as the secondary detection reagent. The fluorescence histograms were derived from gated events based on the light scattering characteristics of viable LS 174T cells. Flow cytometry was performed on a BD FACSCanto™ II Flow Cytometry System.

Product Details
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BD Pharmingen™
LGR5; FEX; HG38; GPR49; GPR67; GRP49
Human (QC Testing)
Rat IgG2b, λ
Human LGR5 DNA
Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence (Not Recommended)
0.5 mg/ml
8549
AB_2737887
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562913 Rev. 2
Antibody Details
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4D11F8

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, 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 organoid 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 4D11F8 monoclonal antibody recognizes an epitope in the center of the leucine-rich repeat (LRR) region of Human Lgr5.

562913 Rev. 2
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
562913 Rev.2
Citations & References
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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. (Biology). 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
562913 Rev. 2

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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.