Anti-GPR30 Polyclonal Antibody 2 is a Rabbit antibody targeting GPR30. Anti-GPR30 Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog)

1. Paraformaldehyde-fixed, paraffin embedded (rat 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-00793) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. 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-00793) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. 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-00793) 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.
4. Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 μg
Lane 2: Cerebrum (Rat) Lysate at 40 μg
Lane 3: HepG2 (Human) Cell Lysate at 30 μg
Lane 4: SH-SY5Y (Human) Cell Lysate at 30 μg
Lane 5: MCF-7 (Human) Cell Lysate at 30 μg
Lane 6: Heart (Mouse) Lysate at 40 μg
Lane 7: Heart (Rat) Lysate at 40 μg
Primary: Anti-GPR30 (TMAB-00793) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
5. Blank control (blue line): A431 cells (fixed with 70% methanol (Overnight at 4°C) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C).
Primary Antibody (green line): Rabbit Anti-GPR30 antibody (TMAB-00793), 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.
6. 25 μg total protein per Lane of various lysates probed with GPR30 polyclonal antibody, unconjugated (TMAB-00793) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

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