Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat

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,IHC-P,IHC-Fr,IF,FCM

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

14160

GPCR,GPCR,Surface Molecules