Anti-GPR30 Polyclonal Antibody is a Rabbit antibody targeting GPR30. Anti-GPR30 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog)

1. Tissue/cell: human colon carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-GPR30 Polyclonal Antibody, Unconjugated 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1:200 dilution for 40 minutes at 37°C.
2. Tissue/cell: human rectal 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-GPR30 Polyclonal Antibody, Unconjugated (TMAB-00792) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control (blue line): A431 cells (blue).
Primary Antibody (green line): Rabbit Anti-GPR30 antibody (TMAB-00792)
Dilution: 1 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% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. 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.
4. Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-GPR30 antibody (TMAB-00792)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
5. Tissue/cell: SH-SY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (GPR30) polyclonal Antibody, Unconjugated (TMAB-00792) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); 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 (GPR30) Polyclonal Antibody, Unconjugated (TMAB-00792) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. 25 μg total protein per Lane of various lysates probed with GPR30 polyclonal antibody, unconjugated (TMAB-00792) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

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

Polyclonal

KLH conjugated synthetic peptide: human GPR30

Human

GPCR,More GPCR,Energy Metabolism,Hormone biosynthesis,Estrogen