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S1PR1 / Sphingosine 1-phosphate receptor 1 · Western blot design guide
Design a Western Blot for S1PR1
Real validated S1PR1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-S1PR1 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
S1PR1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~42.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated S1PR1 Western Blot Protocols
The A01502 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse thymus lysate (catalog A01502) |
| 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 | A01502 · (A) 1 and (B) 2 μg/mL (catalog A01502) |
| 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 S1PR1 Western Blot Band Size?
S1PR1 has a predicted mass of 42.8 kDa; its N-linked glycosylation sites could affect migration, but no visible shift or empirical band size is established.
| Band near 42.8 kDa | Consistent with the predicted S1PR1 mass; confirm identity with band controls |
| Band above 42.8 kDa | N-linked glycans at Asn30 or Asn36 could contribute, but the cause is unconfirmed |
| Little or no band in a soluble lysate fraction | S1PR1 is a multi-pass membrane protein that may be poorly recovered there |
| Several closely spaced bands | Different glycosylation states are possible but have not been shown to resolve on a blot |
💡Expected S1PR1 appearanceUniProt predicts 42.8 kDa for S1PR1; no empirical band size is supplied, and any shifted band needs identity controls because the effect of its glycosylation sites on migration is unproven.
| UniProt predicted mass | Sets a 42.8 kDa sequence-based reference |
| N-linked glycosylation site at Asn30 | May raise apparent mass if occupied; a visible shift is unproven |
| N-linked glycosylation site at Asn36 | May raise apparent mass if occupied; a visible shift is unproven |
| Combined N-linked glycosylation sites at Asn30 and Asn36 | Their combined effect on migration has not been established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated S1PR1 may be poorly recovered during extraction | Check a membrane-enriched fraction and confirm extraction with a membrane-protein control |
| Band higher than expected | N-linked glycosylation at Asn30 or Asn36 is possible | Compare treated and untreated samples and verify band identity |
| Band lower than expected | A specific smaller S1PR1 product is not established by the supplied features | Check band identity with an independent antibody or S1PR1 depletion |
| Broad smear instead of sharp band | Variable glycosylation is possible but has not been demonstrated here | Compare treated and untreated samples and check specificity with S1PR1 depletion |
| Multiple bands | Different glycosylation states are possible; no distinct isoforms are listed | Use an independent antibody or S1PR1 depletion to identify specific bands |
| Weak or no signal | S1PR1 can internalize after ligand binding, and interaction with CD69 promotes its degradation | Check extraction and sample conditions, then use a positive control |
Experiment evidence
Western blot images showing S1PR1 band
A01502 · S1PR1
M01502 · S1PR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for S1PR1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for S1PR1 in Western blot, you can use adrenal gland tissue, where HPA reports high expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As S1PR1 is a membrane protein, a membrane-enriched sample may improve detection; adipose tissue offers an HPA not-detected comparison.
HPA tissue expression evidence for S1PR1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Bergmann glia - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced S1PR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for S1PR1, answered from its protein features.
How should S1PR1 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.
Do documented isoforms explain multiple S1PR1 bands?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The supplied features therefore do not support assigning additional bands to documented S1PR1 isoforms.
Could glycosylation affect the S1PR1 band?
PTM · UniProt lists N-linked glycosylation at Asn30 and Asn36. These sites make glycosylation relevant when assessing an unexpected band, but the features do not establish a visible shift.
UniProt positions are Lys10 (N6-acetyllysine), Thr236 (phosphorylated by PKB/AKT1), and Ser351 and Ser353 (phosphoserines). These coordinates use UniProt numbering and may differ from paper or antibody numbering. Their presence does not establish a visible band shift.
UniProt positions are Lys10 (N6-acetyllysine), Thr236 (phosphorylated by PKB/AKT1), and Ser351 and Ser353 (phosphoserines). These coordinates use UniProt numbering and may differ from paper or antibody numbering. Their presence does not establish a visible band shift.
Does this guide establish induction of S1PR1?
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 S1PR1 Western blot?
Transfer · S1PR1 is a multi-pass membrane protein. Check transfer efficiency for the 42.8 kDa predicted protein with a total-protein stain. The supplied features do not establish whether wet or semi-dry transfer performs better.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01502 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 S1PR1 be quantified across samples?
Quantitation · Use matched sample fractions and normalize to total protein. S1PR1 is listed at the cell membrane and in endosomes; ligand-driven internalization can change its distribution. CD69 interaction promotes degradation, so a lower signal can also reflect reduced receptor abundance.
Should S1PR1 run at its predicted 42.8 kDa?
Interpretation · 42.8 kDa is the predicted mass; no observed band position is supplied. The listed modifications alone cannot establish an apparent mass or explain a difference from 42.8 kDa.
Ligand binding leads to S1PR1 internalization, so compare the same sample fraction across conditions. CD69 interaction promotes S1PR1 degradation and could reduce the signal. The supplied features give no magnitude or timing for either effect.
Compare them with the predicted 42.8 kDa mass, the two N-linked glycosylation sites, and the listed modified residues. One isoform is documented. Without an observed band position or additional evidence, the supplied features cannot identify an unexpected band.
Ligand binding leads to S1PR1 internalization, so compare the same sample fraction across conditions. CD69 interaction promotes S1PR1 degradation and could reduce the signal. The supplied features give no magnitude or timing for either effect.
Compare them with the predicted 42.8 kDa mass, the two N-linked glycosylation sites, and the listed modified residues. One isoform is documented. Without an observed band position or additional evidence, the supplied features cannot identify an unexpected band.
Boster reagents
S1PR1 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-S1P1 S1PR1 Antibody
Cat # A01502
Real WB data
Anti-EDG1 S1PR1 Rabbit Monoclonal Antibody
Cat # M01502
Two the supplier anti-S1PR1 antibodies have Western blot images. A01502 is shown with mouse thymus lysate at 1 and 2 μg/mL. M01502 is shown with human cell, rat brain/C6, and mouse brain/Neuro-2a lysates, with a reported 43 kDa band.
Which to pick: Choose A01502 for a mouse thymus precedent. Choose M01502 for the pictured human cell, rat brain/C6, or mouse brain/Neuro-2a samples. Both list Human, Mouse, and Rat reactivity; the images document specific samples and conditions, not universal performance.
Source: BosterBio S1PR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P21453.
- Human Protein Atlas. S1PR1 tissue expression.
- PMC9613656 — target-verified WB comparison
- PMC5940258 — target-verified WB comparison
- PMC6437142 — target-verified WB comparison