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BD Pharmingen™ Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR)
Clone 4D11F8 (also known as 4D11) (RUO)
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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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 Alexa Fluor® 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; dashed line histogram) or Alexa Fluor® 647 Rat anti-Human Lgr5 (Central LRR) antibody (Cat. No. 562912; solid line histogram) at matched concentrations. 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
BD Pharmingen™
LGR5; FEX; HG38; GPR49; GPR67; GRP49
Human (QC Testing)
Rat IgG2b, λ
Human LGR5 DNA
Flow cytometry (Routinely Tested)
5 µl
AB_2737886
Aqueous buffered solution containing BSA, protein stabilizer, and ≤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 to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Accutase is a registered trademark of Innovative Cell Technologies, Inc.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Alexa Fluor® 647 Rat IgG2b, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557691
Accutase™ Cell Detachment Solution RUO
Size
100 ML
Cat No.
561527
562912 Rev. 2
Antibody Details
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.
562912 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
653 nm
669 nm
562912 Rev.2
Citations & References
View product citations for antibody "562912" on CiteAb
Development References (8)
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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
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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
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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
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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
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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
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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
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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
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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
562912 Rev. 2
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.