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Purified Rat anti-Human Lgr5 (N-Terminal)
Purified Rat anti-Human Lgr5 (N-Terminal)
Western Blot analysis of human LGR5. Lysates from human epithelial kidney cells, HEK293T, that were transiently transfected with empty vector (lane 1) or human LGR5 (lane 2) were probed with Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731) at 2 µg/ml. The second-step reagent was HRP Goat Anti-Rat Ig (Cat. No. 554017). ECL was used for visualization. LGR5 is identified as a band of ~110 kDa. Data generated by Dr. Hans Clevers' Laboratory, Hubrecht Institute.  
Purified Rat anti-Human Lgr5 (N-Terminal)
Flow cytometric analysis of human LGR5. Colorectal adenocarcinoma cells LS 174T (ATCC CL-188) transfected with LGR5 were stained with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 553986, dashed line histogram) or Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731, solid line histogram) at matched concentrations. Cells were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527).The second step reagent was PE Goat Anti-Rat Ig (Cat. No. 550767). The histograms were derived from gated events based on light-scattering characteristics of the LS 174T cells. Flow cytometry was performed on a BD FACSCanto™ II.
Purified Rat anti-Human Lgr5 (N-Terminal)
Immunoflourescent staining of human LGR5. Colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) were fixed and stained with Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731) (pseudo colored green) at 2.5 µg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-rat Ig (Life Technologies) and counter-staining was with DAPI Solution (Cat. No. 564907, pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
Western Blot analysis of human LGR5. Lysates from human epithelial kidney cells, HEK293T, that were transiently transfected with empty vector (lane 1) or human LGR5 (lane 2) were probed with Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731) at 2 µg/ml. The second-step reagent was HRP Goat Anti-Rat Ig (Cat. No. 554017). ECL was used for visualization. LGR5 is identified as a band of ~110 kDa. Data generated by Dr. Hans Clevers' Laboratory, Hubrecht Institute.  
Flow cytometric analysis of human LGR5. Colorectal adenocarcinoma cells LS 174T (ATCC CL-188) transfected with LGR5 were stained with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 553986, dashed line histogram) or Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731, solid line histogram) at matched concentrations. Cells were harvested with Accutase™ Cell Detachment Solution (Cat. No. 561527).The second step reagent was PE Goat Anti-Rat Ig (Cat. No. 550767). The histograms were derived from gated events based on light-scattering characteristics of the LS 174T cells. Flow cytometry was performed on a BD FACSCanto™ II.
Immunoflourescent staining of human LGR5. Colorectal adenocarcinoma cells LS 174T transfected with human LGR5 (Cells from Dr. Hans Clevers, Hubrecht Institute) were fixed and stained with Purified Rat anti-Human Lgr5 (N-Terminal) (Cat. No. 562731) (pseudo colored green) at 2.5 µg/mL. The second-step reagent was Alexa Fluor® 488 goat anti-rat Ig (Life Technologies) and counter-staining was with DAPI Solution (Cat. No. 564907, pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
製品詳細
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BD Pharmingen™
GPR49, GPR67, HG38
Human (QC Testing), Mouse (Lack of Reactivity Confirmed in Development)
Rat IgG2b, κ
Human LGR5
Flow cytometry (Routinely Tested), Bioimaging, Immunofluorescence, Western blot (Tested During Development), Immunohistochemistry-formalin (antigen retrieval required), Immunohistochemistry-frozen (Not Recommended)
~110 kDa
0.5 mg/ml
AB_2737752
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


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. 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.
  6. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562731 Rev. 4
抗体の詳細
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8F2

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 8F2 monoclonal antibody recognizes an epitope in the N-terminal region of Human Lgr5.

562731 Rev. 4
フォーマットの詳細
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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
562731 Rev.4
引用&参考文献
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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
すべて表示する (8) 表示項目を減らす
562731 Rev. 4

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