Leu Rich Repeat containing GPCR

Alternate Nomenclature/Accession # CAI15929, gonadotropin receptor

Relaxin family peptides which closely resembles insulin, act on a group of G-protein-coupled receptors now known as relaxin family peptide (RXFP) receptors. The leucine-rich repeat containing RXFP1 and RXFP2 and the small peptide-like RXFP3 and RXFP4 are the physiological targets for relaxin, insulin-like (INSL) peptide 3, relaxin-3 and INSL5, respectively.  Leucine rich repeat containing GPCR (Lgr6) is a member of the same large family of GPCR (Lgr1-Lgr16) that binds to variety of glycoprotein hormones like TSH, FSH, LH (1). This family of receptor is unique in having a large ectodomain important for ligand binding.  The large extracelluar domain binds the ligands with high affinity; there also exist a low affinity binding site in the exoloop of the trnasmembrane region.  LGR6 has a subgroup-specific hinge region after leucine-rich repeats whereas LGR7, like snail LGR, contains a low density lipoprotein (LDL) receptor cysteine-rich motif at the N terminus.  The expression of LGR5 is induced in the limb, tail, and intestine by T3 during metamorphosis of frog form larval stage. More importantly, LGR5 mRNA is localized to the developing adult epithelial stem cells of the intestine (2). Lgr6 are also involved in transcription silencing of CpG islands by hypermethylation in colorectal cancer which are referred as methylator phenotypes. The CAN genes include SYNE1, STRAD8, CD109, RNF, RET, APC2 MMP2, PTPRD and Lgr6 (3). 

Lgr5 is a marker for cycling stem cells in intestine (4) whereas Lgr6  is expressed in earliest embryonic hair placodes (5).Lgr6 positive cells are also involved in wound healing.   In adult hair follicles the Lgr6 resides in a region just above the follicle bulge, this region was previously uncharacterized and does not express any known bulge stem cell markers.  The Lgr6 is a marker for a distinct population of stem cells that give rise to all lineages of the skin.  Unlike Lgr5, the Lgr6 is not regulated by Wnt signaling. 

The Lgr6-selective antibodies were generated against antigenic peptide taken from human Lgr6 protein corresponding near to carboxy terminal Lgr6 protein.  Lgr6-antibodies are affinity purified over immobilized antigen based chromatography, and the purified immunoglobulins are stabilized in antibody stabilization buffer. FabGennix Int. Inc., will also provide limited quantities of antigenic blocking protein for Lgr6-101AP antibody. Antibodies to Lgr6 (Lgr-601AP) labels ~115-118kDa protein in PC-Lgr6 samples. The Lgr6 antibodies can be labeled (FITC, Biotin, secondary enzymes etc.) upon request at a nominal charge.   FabGennix has a wide range of antibodies for epigenetics, cell biology, neurosciences an dsignal transduction research, for a complete listing please visit www.FabGennix.com. 

Immunogen:            Synthetic peptides corresponding to unique epitope on Lgr6.  The peptide sequence corresponding to amino acids 860-915 of the human Lgr6 protein, the peptide was post-synthetically modified to achieve desired antigenicity before coupling to a carrier protein.      

Concentration:         Lgr6-101AP:  IgG concentration: 0.58-0.65 mg/ml in antibody stabilization buffer.

Applications:           Antibody Lgr6 -101AP is ideal for WB and ELISA applications, other applications have not been tested. The dilutions for this antibody is for reference only, investigators are expected to determine the optimal conditions for specific assay.  WB: > 1:       IMM & i.p pull-down assays: n.d; IHC n.d.  ELISA <1:10,000.  Application of this antibody in protocols not listed here or not determined (“n.d”) does not necessarily exclude its use in such procedures. For standardization of such protocols, FabGennix will be provide “Free Samples”.  Please contact technical service for more details.

Reactivity:               This antibody detects a fuzzy band of approximately 115 kDa in PC-Lgr6 samples and few additional bands between MW45-60kDa were also observed.  The identity of these bands are not known but could be proteolytic fragment of the full-length Lgr6  molecule.

Protocols:                Standard protocol for various applications (WB; IMM and IHC) of this antibody is provided with the product specification sheet, however, FabGennix Inc., strongly recommends investigators to optimize conditions for use of this antibody in their laboratories.  

Form/Storage:         The antiserum is supplied in antibody stabilization buffer.  The affinity-purified antibodies are isolated on immobilized antigen-affinity column and supplied as stabilized product.  Store at -20^oC for long-term storage.  FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45um filter after every use for long-term storage.

 

Notes:  Briefly centrifuge to collect liquid, heat or boil PC-Lgr6  tube for 1-2 minutes to dissolve any precipitate before use.  This product is “ready-to-use” for electrophoresis. After thawing store at room temperature, Repeated freezing and thawing may result in appearance of higher MW immunoreactive bands.   

 

New Reagents:  Now you can recycle your western blots (nitrocellulose, supported membranes and PVFD membranes) by using our StripObuffer (Cat FGI-1989).  Each stripping is guaranteed to give better signal (up to 8 stripping).  No strong pungent smell of reducing agents or heating required.  Block in 5X diluObuffer and you are ready for blotting with a new antibody

 

 

For research use only, not for diagnostic or therapeutic use.