GPR30 Rabbit Polyclonal Antibody

• Catalog Number: orb10740
• Category: Antibodies
• Description: GPR30 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Canine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GPR30 (251-375/375aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: GPER1
• UniProt ID: Q99527
• RRID: AB_10747838
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: G Protein Coupled Receptor 30; G-protein coupled r
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Ebrahim Elsangeedy * 1,4 , Dina N. Yamaleyeva * 1,4 , Nicholas P. Edenhoffer 3,4 , Allyson Deak 1,4 , Anna Soloshenko 1,4 , Jonathan Ray 1,4 , Xuming Sun 1,4 , Omar H. Shaltout 1,4 , Nildris Cruz Diaz 1,4 , Brian Westwood 1,4 , Daniel Kim-Shapiro 5 , Debra I. Diz 1,4 , Shay Soker 3,4 , Victor M. Pulgar 1,4,6 , April Ronca 4,7 , Jeffrey S. Willey 2,4 , and Liliya M. Yamaleyeva Sex-Specific Cardiovascular Adaptations to Simulated Microgravity in Sprague-Dawley Rats bioRxiv, (2024)

PubMed:

Applications

IHC

Reactivity

Rat

Kalabay, Márton et al. Investigation of the Antitumor Effects of Tamoxifen and Its Ferrocene-Linked Derivatives on Pancreatic and Breast Cancer Cell Lines Pharmaceuticals (Basel), 15, 314 (2022)

PubMed: 35337112

Blank control (blue line): A431 cells (blue). Primary Antibody (green line): Rabbit Anti-GPR30 antibody (orb10740), 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. Acquisition of 20000 events was performed.

Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-GPR30 antibody (orb10740), 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. Acquisition of 20000 events was performed.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GPR30) Polyclonal Antibody, Unconjugated (orb10740) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat uterus), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GPR30) Polyclonal Antibody, Unconjugated (orb10740) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Heart (mouse) Lysate at 40 ug, Primary: Anti-GPR30 (orb10740) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 50kD.

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 (orb868589) used at 1:200 dilution for 40 minutes at 37°C.

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 (orb10740) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

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 (orb10740) 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 nuclei.

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 (orb10740) 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 nuclei.