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- Table of Contents
GPER1 / G-protein coupled estrogen receptor 1 · Western blot design guide
Design a Western Blot for GPER1
Source-linked GPER1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GPER1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
GPER1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~42.2 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Cervix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked GPER1 Western Blot Protocol Options
The PA2235 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | COLO320, MCF-7, COS7 (catalog PA2235) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | PA2235; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected GPER1 Western Blot Band Size?
GPER1 has a predicted monomer mass of 42.2 kDa; N-linked glycans and a probable homodimer could affect migration, but no empirical band size is supplied.
| Band near 42.2 kDa | consistent with the predicted GPER1 monomer; confirm identity with antibody controls |
| Band above 42.2 kDa | could reflect N-linked glycosylation at Asn25, Asn32, or Asn44 |
| Band near twice the monomer size | could reflect a retained homodimer, which UniProt describes as probable |
| Weak band in whole-cell lysate | could reflect incomplete recovery of membrane-associated GPER1 |
💡Expected GPER1 appearanceUniProt predicts a 42.2 kDa GPER1 monomer; no empirical band size is supplied, and glycosylation or retained homodimers could alter migration, so confirm band identity with appropriate controls.
| Predicted monomer mass | 42.2 kDa is the sequence-based reference size |
| N-linked glycosylation at Asn25 | could increase apparent mass if the site carries a glycan |
| N-linked glycosylation at Asn32 | could increase apparent mass if the site carries a glycan |
| N-linked glycosylation at Asn44 | could increase apparent mass if the site carries a glycan |
| Probable homodimer | could produce a band near twice the monomer size if the complex survives sample preparation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane-associated GPER1 may be poorly recovered | check membrane extraction and use a positive-control lysate |
| Band higher than expected | N-linked glycosylation or a retained homodimer could contribute | compare glycosidase-treated samples and reducing sample conditions |
| Band lower than expected | the supplied features do not establish a specific smaller GPER1 species | check band identity with an independent antibody or GPER1-depleted control |
| Broad smear instead of sharp band | different glycan states at the listed N-linked sites are possible | compare untreated and glycosidase-treated samples |
| Multiple bands | glycan states or retained receptor complexes could differ | compare glycosidase treatment and reducing conditions, then confirm identity with a GPER1-depleted control |
| Weak or no signal | incomplete solubilization of membrane-associated GPER1 | check the membrane-enriched fraction and a positive-control lysate |
Experiment evidence
Western blot images showing GPER1 band
PA2235 · GPER1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GPER1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GPER1 in Western blot, you can use appendix tissue, which has high HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Cervix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: GPER1 is a multi-pass membrane protein, so membrane-enriched lysate may improve detection.
HPA tissue expression evidence for GPER1
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced GPER1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GPER1, answered from its protein features.
How should GPER1 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could annotated isoforms explain multiple GPER1 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no annotated isoform explanation for multiple bands. Assess band identity with appropriate antibody and sample controls.
Which GPER1 glycosylation sites matter when assessing band heterogeneity?
PTM · UniProt lists N-linked glycosylation at Asn25, Asn32, and Asn44. Use these UniProt coordinates when describing candidate glycosylated forms; check the numbering convention before comparing a paper or antibody datasheet. The annotations alone cannot identify the cause of multiple bands.
UniProt lists N-acetylmethionine at position 1. Keep that UniProt coordinate when comparing modification reports. This annotation alone does not establish a detectable Western blot shift.
UniProt lists N-acetylmethionine at position 1. Keep that UniProt coordinate when comparing modification reports. This annotation alone does not establish a detectable Western blot shift.
Does this guide establish induction of GPER1?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for GPER1 Western blot?
Transfer · GPER1 is a 375-residue, multi-pass membrane protein with a predicted mass of 42.2 kDa. Choose transfer conditions that recover proteins around this mass, and check transfer with a total-protein stain. The supplied features do not specify wet or semi-dry transfer settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PA2235 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should GPER1 be quantified across sample fractions?
Quantitation · GPER1 is annotated at cell and vesicle membranes, endosomes, Golgi, cytoplasm, and nucleus. Compare like fractions across samples and normalize each lane to its measured protein input. A change in one fraction need not represent a change in total cellular GPER1.
Should GPER1 migrate at its predicted 42.2 kDa?
Interpretation · 42.2 kDa is the predicted mass, and no observed band size was supplied. GPER1 has three annotated N-linked glycosylation sites, but their presence alone does not establish a visible shift or explain any difference from 42.2 kDa.
GPER1 is annotated as a probable homodimer and as forming heterodimers with other GPCRs, including CRHR1, HTR1A, and PAQR8. Those annotations make complexes worth considering, but they do not identify an unexpected band. Check band identity and whether its appearance depends on sample preparation.
GPER1 is annotated as a probable homodimer and as forming heterodimers with other GPCRs, including CRHR1, HTR1A, and PAQR8. Those annotations make complexes worth considering, but they do not identify an unexpected band. Check band identity and whether its appearance depends on sample preparation.
Boster reagents
GPER1 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-G-protein coupled receptor 30/GPER1 Antibody Picoband®
Cat # PA2235
PA2235 is an anti-GPER1 antibody listed for human and monkey reactivity. Its Western blot image shows COLO320, MCF-7, and COS7 cell lysates. The supplied evidence does not establish performance across other samples or conditions.
Which to pick: PA2235 is the only listed GPER1 antibody and has a Western blot image. Check its listed human and monkey reactivity and the three imaged cell lysates against your planned sample.
Source: BosterBio GPER1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99527.
- Human Protein Atlas. GPER1 tissue expression.
- PMC6598863 — target-verified WB comparison
- PMC7793665 — target-verified WB comparison
- PMC8235990 — target-verified WB comparison