Anti-LGR-5 Polyclonal Antibody is a Rabbit antibody targeting LGR-5. Anti-LGR-5 Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse,Rat

1. Sample:
Brain (Mouse) Lysate at 40 μg
Primary: Anti-GPR49 (TMAB-01063) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 98 kDa
2. Sample:
LOVO (Human) Cell Lysate at 30 μg
HepG2 (Human) Cell Lysate at 30 μg
Primary: Anti-GPR49 (TMAB-01063) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 125 kDa
3. Tissue/cell: human colon carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-GPR49/LGR5 Polyclonal Antibody, Unconjugated (TMAB-01063) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Tissue/cell: mouse colon carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-GPR49/LGR5 Polyclonal Antibody, Unconjugated (TMAB-01063) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
5. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-GPR49 antibody (TMAB-01063),
Dilution: 3 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
Protocol
The cells were fixed with 70% ethanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.
6. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (GPR49) Polyclonal Antibody, Unconjugated (TMAB-01063) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: mouse GPR49

Mouse

LGR5

Leucine-rich repeat-containing G-protein coupled receptor 5

GPCR,GPCR,Surface Molecules

Receptor for R-spondins that potentiates the canonical Wnt signaling pathway and acts as a stem cell marker of the intestinal epithelium and the hair follicle. Upon binding to R-spondins (RSPO1, RSPO2, RSPO3 or RSPO4), associates with phosphorylated LRP6 and frizzled receptors that are activated by extracellular Wnt receptors, triggering the canonical Wnt signaling pathway to increase expression of target genes. In contrast to classical G-protein coupled receptors, does not activate heterotrimeric G-proteins to transduce the signal. Involved in the development and/or maintenance of the adult intestinal stem cells during postembryonic development.