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- Table of Contents
PTGER2 / Prostaglandin E2 receptor EP2 subtype · Western blot design guide
Design a Western Blot for PTGER2
Source-linked PTGER2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTGER2 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
PTGER2 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 | ~39.8 kDa | |
| Observed band | 40 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked PTGER2 Western Blot Protocol Options
The PB9386 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human Placenta at 50ug, A549 at 40ug, HEPG2 at 40ug Predicted bind size: 40KD Observed bind size: 40KD (catalog PB9386) |
| 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 | PB9386; 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 PTGER2 Western Blot Band Size?
PTGER2 is predicted at 39.8 kDa and observed at 40 kDa; the cause of their slight difference is not established.
| Band near 40 kDa | Matches the empirical PTGER2 band and its 39.8 kDa predicted mass; confirm identity with antibody controls |
| Band above 40 kDa | Could reflect N-linked glycosylation at annotated sites, though a visible shift is not established |
| Diffuse signal near 40 kDa | Could reflect variable N-linked glycosylation; the supplied evidence does not demonstrate a smear |
| Weak band in whole-cell lysate | Could reflect limited recovery of this multi-pass membrane protein |
💡Expected PTGER2 appearanceThe empirical PTGER2 band is 40 kDa in the supplied blots, close to UniProt's 39.8 kDa predicted mass; use antibody controls to confirm band identity.
| Predicted PTGER2 mass | 39.8 kDa predicted; the empirical band is 40 kDa |
| N-linked glycosylation at Asn3 | Could alter apparent mass if occupied; no site-specific shift is established |
| N-linked glycosylation at Asn6 | Could alter apparent mass if occupied; no site-specific shift is established |
| N-linked glycosylation at Asn96 | Could alter apparent mass if occupied; no site-specific shift is established |
| N-linked glycosylation at Asn287 | Could alter apparent mass if occupied; no site-specific shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PTGER2 is a multi-pass membrane protein that may be poorly recovered | Check membrane extraction and compare with the validated lysates |
| Band higher than expected | N-linked glycosylation may affect migration; the shift is unproven | Compare with a validated positive control and assess glycosylation if needed |
| Band lower than expected | The supplied features do not establish a smaller PTGER2 product | Check antibody specificity and compare with the 40 kDa positive-control band |
| Broad smear instead of sharp band | Variable glycosylation is possible but not demonstrated | Check sample preparation and compare with a validated positive control |
| Multiple bands | The supplied features do not establish distinct PTGER2 isoform bands | Confirm which band tracks with the 40 kDa positive control |
| Weak or no signal | Recovery of membrane-localized PTGER2 may be limited | Check extraction and loading against a validated positive control |
Experiment evidence
Western blot images showing PTGER2 band
PB9386 · PTGER2
M04963 · PTGER2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PTGER2 Western blot
🧪For positive controls for PTGER2 in Western blot, you can use an HPA-validated tissue or cell line once identified; the supplied evidence names none.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: PTGER2 is a membrane protein, and the supplied HPA data identify no validated positive or negative tissue.
HPA tissue expression evidence for PTGER2
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced PTGER2 Western Blot Tips
Deeper troubleshooting and optimisation questions for PTGER2, answered from its protein features.
How should PTGER2 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 PTGER2 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore does not support assigning additional bands to annotated isoforms. Check band identity before interpreting multiple bands as PTGER2.
Which PTGER2 glycosylation sites matter when interpreting bands?
PTM · UniProt annotates N-linked glycosylation at positions 3, 6, 96, and 287, using its supplied sequence numbering. Keep those coordinates when comparing records or antibody information; other numbering conventions may differ. The annotations do not establish which sites are occupied in a particular sample or whether glycosylation changes its apparent mass.
Does this guide establish induction of PTGER2?
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 PTGER2 Western blot?
Transfer · PTGER2 is a 358-residue, multi-pass cell-membrane protein with a predicted mass of 39.8 kDa. The supplied features do not specify a transfer method. Choose conditions that recover a membrane protein near 40 kDa, and verify transfer in the relevant region.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PB9386 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 PTGER2 bands be quantified?
Quantitation · Quantify the same band region near the supplied observed 40 kDa across samples, using consistent membrane-protein preparation and a suitable loading reference. PTGER2 is a multi-pass membrane protein, so inconsistent recovery could affect comparisons. Do not count extra bands as PTGER2 without establishing their identity.
Why is the PTGER2 band near 40 kDa?
Interpretation · UniProt predicts 39.8 kDa, and the supplied observed band is 40 kDa. That agreement needs no band-shift explanation. PTGER2 has four annotated N-linked glycosylation sites, but their presence alone does not establish a visible shift.
Start with the expected region near 40 kDa. The record lists four N-linked glycosylation sites and one disulfide bond, but neither feature alone explains an additional band or proves a shift. It lists no alternative sequence or oligomer assignment; establish the identity of unexpected bands before attributing them to PTGER2.
UniProt lists one disulfide bond, so record whether samples were run under reducing or nonreducing conditions when comparing blots. The supplied features give no bond coordinates or measured mobility effect; do not assign a band difference to that bond without supporting evidence.
Start with the expected region near 40 kDa. The record lists four N-linked glycosylation sites and one disulfide bond, but neither feature alone explains an additional band or proves a shift. It lists no alternative sequence or oligomer assignment; establish the identity of unexpected bands before attributing them to PTGER2.
UniProt lists one disulfide bond, so record whether samples were run under reducing or nonreducing conditions when comparing blots. The supplied features give no bond coordinates or measured mobility effect; do not assign a band difference to that bond without supporting evidence.
Boster reagents
PTGER2 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-Prostaglandin E Receptor EP2/PTGER2 Antibody Picoband®
Cat # PB9386
Real WB data
Anti-PTGER2 Monoclonal Antibody
Cat # M04963
The catalog reports two anti-PTGER2 antibodies with Western blot images. PB9386 shows a 40 kDa band in human samples; M04963 shows an approximately 52 kDa band against a 40 kDa expectation. The supplied evidence is limited to product captions.
Which to pick: For human samples, PB9386 has an image showing placenta, A549, and HEPG2 lysates. M04963 includes human cell lines plus rat spleen and mouse lung and spleen, but its observed band differs from the expected size. Choose based on your sample and band interpretation.
Source: BosterBio PTGER2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P43116.
- Human Protein Atlas. PTGER2 tissue expression.
- PMC13385061 — target-verified WB comparison
- PMC2714846 — target-verified WB comparison
- PMC9504288 — target-verified WB comparison