Description
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.
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.
1. 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).
2. An isotype control should be used at the same concentration as the antibody of interest.
3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
4. 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.
5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. 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.
8. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
9. Accutase is a registered trademark of Innovative Cell Technologies, Inc.
10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.