![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - The MW of GPR30 is estimated to be 42 kDa, but higher MW sizes have been reported due to glycosylation and interaction with other proteins. Bands are identified as glycosylated and nonglycosylated based on reports cited in the text, but could include interaction with other proteins. For each GPER western, the membrane was stripped and reprobed for b-actin as a loading control. Quantitation of GPER was evaluated by summing all immunoreactive bands and dividing by b-actin, then normalizing to HBEC2-KT in each blot. Image collected and cropped by CiteAb from the following publication (biomedcentral.com/1471-2407/12/624), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/GPER-GPR30-Antibody-Western-Blot-NBP1-31239-img0012.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - The MW of GPR30 is estimated to be 42 kDa, but higher MW sizes have been reported due to glycosylation and interaction with other proteins. Bands are identified as glycosylated and nonglycosylated based on reports cited in the text, but could include interaction with other proteins. For each GPER western, the membrane was stripped and reprobed for b-actin as a loading control. Quantitation of GPER was evaluated by summing all immunoreactive bands and dividing by b-actin, then normalizing to HBEC2-KT in each blot. Image collected and cropped by CiteAb from the following publication (biomedcentral.com/1471-2407/12/624), licensed under a CC-BY license.")
| Reactivity | Hu, Po, RtSpecies Glossary |
| Applications | WB, Flow, ICC/IF, IHC, KD |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Format | BSA Free |
| Immunogen | Carrier-protein conjugated synthetic peptide encompassing a sequence within the C-terminus region of human GPER/GPR30. The exact sequence is proprietary. |
| Localization | Cell membrane, Multi-pass membrane protein, Endoplasmic reticulum membrane, Multi-pass membrane protein, Golgi apparatus membrane, Multi-pass membrane protein |
| Predicted Species | Rat (92%). Backed by our 100% Guarantee. |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | GPER1 |
| Purity | Antigen Affinity-purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| Application Notes | WB As is commonly seen with membrane proteins, significant hydrophobicity can lead to aggregation following boiling of samples prior to SDS-PAGE and subsequent western blotting. We recommend to avoid boiling in this case. After harvesting lysate with RIPA buffer, sample loading buffer (including 2-ME) is directly added to the lysate. Samples are then mixed well and added directly without heating. |
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| Theoretical MW | 42 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | PBS, 20% Glycerol |
| Preservative | 0.01% Thimerosal |
| Purity | Antigen Affinity-purified |
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| Images | Ratings | Applications | Species | Date | Details | ||||||||
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reviewed by:
Verified Customer |
IF | Human | 03/05/2013 |
Summary
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Secondary Antibodies |
Isotype Controls |
Research Areas for GPER/GPR30 Antibody (NBP1-31239)Find related products by research area.
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![Immunocytochemistry/Immunofluorescence: GPER/GPR30 Antibody [NBP1-31239] - Detection of GPER/GPR30 in HUVEC nucleus. Image courtesy of a product review by Dr. Subhadeep Chakrabarti of University of Alberta.](http://images.novusbio.com/fullsize/GPER-GPR30-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-31239-img0010.jpg "Immunocytochemistry/Immunofluorescence: GPER/GPR30 Antibody [NBP1-31239] - Detection of GPER/GPR30 in HUVEC nucleus. Image courtesy of a product review by Dr. Subhadeep Chakrabarti of University of Alberta.")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - A. 50 ug 293T whole cell lysate/extract. B. 50 ug whole cell lysate/extract of GFP-human GPR30-transfected 293T cells (No boiling). C. 50 ug whole cell lysate/extract of GFP-human GPR30 and GPR30 siRNA-transfected 293T cells (No boiling). 5 % SDS-PAGE GPR30 antibody ) dilution: 1:1000](http://images.novusbio.com/fullsize/GPER-GPR30-Antibody-Western-Blot-NBP1-31239-img0011.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - A. 50 ug 293T whole cell lysate/extract. B. 50 ug whole cell lysate/extract of GFP-human GPR30-transfected 293T cells (No boiling). C. 50 ug whole cell lysate/extract of GFP-human GPR30 and GPR30 siRNA-transfected 293T cells (No boiling). 5 % SDS-PAGE GPR30 antibody ) dilution: 1:1000")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - AhR & GPR30 receptors are present & functional in the MCF10AT1 cells. (A) RT-qPCR analysis of AhR & GPR30 mRNA expression levels represented in arbitrary units (a.u.) in the MCF10AT1 & MCF10CA1a.cl1 cells. MCF-7 cells were used as a control. Values represent mean ± SD of three independent experiments conducted in triplicate. (B) Representative Western blot analyses from three independent experiments of AhR & GPR30 protein expression in MCF10AT1 & MCF10CA1a.cl1 cells. MCF-7 cells were used as a control. (C) XRE-luciferase activity following 8 h exposure of MCF10AT1 cells to ITE at the indicated concentrations. TCDD 10−7 M was used as a control & results were expressed as % of TCDD 10−7 M activity. ***p < 0.001 in Student t-test. (D) XRE-luciferase activity upon 8 h of exposure to ITE 10−10 M alone or in combination with GNF351 at the indicated concentrations. TCDD 10−7 M was used as a control, & results were expressed as % of TCDD 10−7 M activity. Student t-tests revealed the statistically significant differences between unexposed & exposed cells: ***p < 0.001; & between ITE & ITE+GNF351: ###p < 0.001. Values in (C,D) represent mean ± SD of three independent experiments. (E) Representative Western blot analyses from three independent experiments of the phospho-MAPK/MAPK ratio upon exposure of MCF10AT1 cells to G1 (GPR30 agonist) for the times indicated, in the presence or absence of a 2 h pre-treatment with G15 (GPR30 antagonist). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-31239_rabbit-polyclonal-gper-gpr30-antibody-310202416171440.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - AhR & GPR30 receptors are present & functional in the MCF10AT1 cells. (A) RT-qPCR analysis of AhR & GPR30 mRNA expression levels represented in arbitrary units (a.u.) in the MCF10AT1 & MCF10CA1a.cl1 cells. MCF-7 cells were used as a control. Values represent mean ± SD of three independent experiments conducted in triplicate. (B) Representative Western blot analyses from three independent experiments of AhR & GPR30 protein expression in MCF10AT1 & MCF10CA1a.cl1 cells. MCF-7 cells were used as a control. (C) XRE-luciferase activity following 8 h exposure of MCF10AT1 cells to ITE at the indicated concentrations. TCDD 10−7 M was used as a control & results were expressed as % of TCDD 10−7 M activity. ***p < 0.001 in Student t-test. (D) XRE-luciferase activity upon 8 h of exposure to ITE 10−10 M alone or in combination with GNF351 at the indicated concentrations. TCDD 10−7 M was used as a control, & results were expressed as % of TCDD 10−7 M activity. Student t-tests revealed the statistically significant differences between unexposed & exposed cells: ***p < 0.001; & between ITE & ITE+GNF351: ###p < 0.001. Values in (C,D) represent mean ± SD of three independent experiments. (E) Representative Western blot analyses from three independent experiments of the phospho-MAPK/MAPK ratio upon exposure of MCF10AT1 cells to G1 (GPR30 agonist) for the times indicated, in the presence or absence of a 2 h pre-treatment with G15 (GPR30 antagonist). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-31239_rabbit-polyclonal-gper-gpr30-antibody-310202416173515.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Human endothelial cells express GPR30 protein in the cell nucleus.(A) Confluent monolayers of HUVECs from 3 different cords were lysed & the protein lysates were immunoblotted for GPR30. alpha -tubulin was used as loading control. (B) HUVEC monolayers at 30–40% confluence were treated with 40 nM siRNA (control or GPR30_4) for 48 hours prior to lysis followed by immunoblotting of the cell lysates for GPR30. alpha -tubulin was used as loading control. Data shown are mean ± SEM of 4 independent experiments. ** & ## indicate p<0.01 compared to untreated & control siRNA-treated cells, respectively. (C) Confluent HUVECs grown on glass coverslips were fixed, permeabilized & immunostained with anti-GPR30 antibody. Nuclei were stained with Hoechst33342 dye. The merged image shows GPR30 (red) & nuclei (blue) in pseudocolor. Representative images from 3 independent experiments are shown. Bar, 20 µm. (D) Confluent HUVECs were lysed & fractionated into cytosolic (C) & nuclear (N) fractions prior to western blotting for eNOS, GPR30, p65, alpha -tubulin & c-Jun. A representative set of images (obtained from different membranes) from 3 independent experiments is shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23285008), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-31239_rabbit-polyclonal-gper-gpr30-antibody-310202416175294.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Human endothelial cells express GPR30 protein in the cell nucleus.(A) Confluent monolayers of HUVECs from 3 different cords were lysed & the protein lysates were immunoblotted for GPR30. alpha -tubulin was used as loading control. (B) HUVEC monolayers at 30–40% confluence were treated with 40 nM siRNA (control or GPR30_4) for 48 hours prior to lysis followed by immunoblotting of the cell lysates for GPR30. alpha -tubulin was used as loading control. Data shown are mean ± SEM of 4 independent experiments. ** & ## indicate p<0.01 compared to untreated & control siRNA-treated cells, respectively. (C) Confluent HUVECs grown on glass coverslips were fixed, permeabilized & immunostained with anti-GPR30 antibody. Nuclei were stained with Hoechst33342 dye. The merged image shows GPR30 (red) & nuclei (blue) in pseudocolor. Representative images from 3 independent experiments are shown. Bar, 20 µm. (D) Confluent HUVECs were lysed & fractionated into cytosolic (C) & nuclear (N) fractions prior to western blotting for eNOS, GPR30, p65, alpha -tubulin & c-Jun. A representative set of images (obtained from different membranes) from 3 independent experiments is shown. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/23285008), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-31239_rabbit-polyclonal-gper-gpr30-antibody-310202416173516.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/nbp1-31239_rabbit-polyclonal-gper-gpr30-antibody-310202416163721.jpg "Western Blot: GPER/GPR30 Antibody [NBP1-31239] - Effects of short-term exposure of BPA, B[a]P, ITE & G1 10−10 M on AIG & MFE are inhibited by siRNA-AhR & siRNA-GPR30. Representative Western blot analysis from three independent experiments of AhR & GPR30 expression in transfected MCF10AT1 cells with (A) siRNA-AhR, (B) siRNA-GPR30 or their scrambled controls. Quantification of protein expression levels was normalized against tubulin expression. (C,D) Secondary mammospheres formation & (E,F) average number of colonies in soft agar, with the following treatments: BPA and/or B[a]P, G1, or ITE, 10−10 M. Cells were transfected with either siRNA-AhR, siRNA-GPR30 or their scrambled controls before being subjected to the treatments. Treatments were maintained throughout the course of experiments. (mean ± SD of 2 independent experiments, in triplicate). ***p < 0.001, *p < 0.05 vs. their respective unexposed; ###p < 0.001 siRNA vs. scrambled in Student t-test. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32670863), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
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