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S1PR2 / Sphingosine 1-phosphate receptor 2 · IHC design guide
Design Immunohistochemistry for S1PR2
Plan chromogenic S1PR2 IHC in paraffin sections using high staining in duodenal glandular cells as a positive reference and absent staining in adipose tissue adipocytes as a negative reference (HPA tissue IHC). The catalog antibody lists 10 μg/ml for IHC, with an IHC image from mouse brain tissue (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
S1PR2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cell membrane expected (UniProt); microvilli observed (HPA tissue IHC) | |
| Staining pattern | Distinct staining in erythrocytes and intestinal microvilli (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+2 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Erythrocyte staining may complicate scoring (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; check extracellular vs cytoplasmic epitope location (UniProt) |
Section 1
Recommended S1PR2 IHC & IF Protocols
The catalog antibody protocol is paired with published S1PR2 IHC methods for paraffin sections and frozen tissue (PMC4587559; PMC10949307; PMC8972069).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A02904); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-S1PR2, 10 μg/ml (datasheet A02904) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | S1PR2-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: Distinct expression in erythrocytes and intestinal microvilli. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule); use EDTA retrieval when reproducing PMC4587559.
Section 2
What Is the Expected S1PR2 Staining Pattern?
S1PR2 is a seven transmembrane cell membrane receptor (UniProt O95136 topology and subcellular location). In paraffin section IHC, look first for staining along intestinal glandular cell surfaces, especially the microvilli described by HPA; duodenal and small intestinal glandular cells are scored High, and kidney tubular cells Medium (HPA tissue IHC). HPA rates its tissue IHC profile Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Intestinal glandular cells show strong apical or microvillar staining. | This fits HPA's High staining in duodenal and small intestinal glandular cells and its distinct intestinal microvillar pattern (HPA tissue IHC). A membrane associated distribution is consistent with S1PR2's seven transmembrane topology (UniProt O95136 topology). Compare the stained edge with tissue morphology before scoring it. |
| Staining is predominantly nuclear, or fills cells without a discernible membrane associated pattern. | That pattern is discordant with the UniProt cell membrane annotation and should prompt an artefact check (UniProt O95136 subcellular location). HPA reports uncertain nuclear speckles in ICC/IF, but that observation does not establish a nuclear pattern for paraffin section IHC (HPA subcellular ICC/IF). |
| Strong staining appears in a cell population HPA lists as not detected. | For example, HPA scores adipocytes in adipose tissue and respiratory epithelial cells in bronchus as Not detected (HPA tissue IHC). Check morphology and controls before interpreting signal there as S1PR2; antibody cross reactivity or endogenous detection activity are possible explanations (general IHC practice). |
| Color covers stroma, empty spaces, and many unrelated cells with little structure. | A broadly diffuse deposit is difficult to reconcile with the reported intestinal microvillar pattern (HPA tissue IHC). It may reflect background from the antibody or detection system; compare a no primary control and assess blocking and washing before assigning a cellular score (general IHC practice). |
| Duodenal or small intestinal glandular cells have no detectable signal. | These are useful positive reference populations because HPA scores their staining High (HPA tissue IHC). First check that the expected cells and microvilli are present, then review antibody suitability, antigen retrieval, detection, and the control slide (general IHC practice). One negative section alone cannot establish absent S1PR2. |
💡Expected S1PR2 appearanceCall a positive IHC result when intestinal glandular cells show strong, structured membrane associated or microvillar staining (HPA tissue IHC; UniProt O95136 subcellular location); widespread featureless color or dominant nuclear staining warrants an artefact check (general IHC practice).
| Membrane topology | S1PR2 has seven membrane spanning segments and extracellular and cytoplasmic regions (UniProt O95136 topology). Interpret a sharply outlined cell surface or microvillar pattern in that context; topology alone does not specify which epitope the IHC antibody recognizes. |
| Reference tissue and cell choice | HPA scores duodenal and small intestinal glandular cells High and kidney tubular cells Medium (HPA tissue IHC). Use the intestinal cells as the clearer positive reference, while keeping the tissue's cell level annotation in view when comparing sections. |
| Antibody evidence | HPA014307 is Approved for IHC; HPA014255 has no listed IHC status in the supplied antibody record (HPA antibodies). The tissue profile itself has medium staining to RNA consistency (HPA tissue IHC), so interpret a new compartment or cell type with controls rather than treating approval as universal specificity. |
| Erythrocyte signal | HPA describes distinct erythrocyte staining (HPA tissue IHC). Record it separately from glandular cell staining. If using an enzyme based chromogenic system, evaluate whether endogenous activity contributes to blood cell color with an appropriate control (general IHC practice). |
| IF/ICC Q: should the reported intracellular pattern guide this IHC score? | A: HPA places S1PR2 mainly at the Golgi apparatus and additionally at nuclear speckles in ICC/IF, marking both locations uncertain (HPA subcellular ICC/IF). These observations warrant caution when comparing modalities; they do not override UniProt's cell membrane annotation or HPA's tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The intestinal positive reference is blank. | The expected glandular cells may be absent from the section, or staining conditions may have failed; HPA reports High staining in those cells (HPA tissue IHC). | Confirm glandular morphology on the section and inspect the positive control. Review antibody suitability, retrieval, and detection using the established IHC workflow (general IHC practice). |
| Only nuclei are strongly colored. | Dominant nuclear IHC staining conflicts with the cell membrane annotation (UniProt O95136 subcellular location). HPA's nuclear speckle assignment comes from uncertain ICC/IF data (HPA subcellular ICC/IF). | Compare the slide with a no primary control and recheck tissue morphology and antibody specificity before scoring nuclear color as S1PR2 (general IHC practice). |
| Color is widespread and obscures the intestinal brush border. | Diffuse background can obscure the distinct microvillar pattern reported by HPA (HPA tissue IHC; general IHC practice). | Review blocking, washing, antibody concentration, and detection background with matched controls; then reassess whether a structured microvillar edge remains (general IHC practice). |
| An HPA negative population stains as strongly as intestinal glands. | Cell identification may be mistaken, or the signal may reflect cross reactivity or endogenous detection activity (general IHC practice); HPA records several cell populations as Not detected (HPA tissue IHC). | Verify the cell type and compare its signal with a no primary control. Keep cell specific scores separate rather than assigning the whole tissue one result (general IHC practice). |
| Erythrocytes dominate the chromogenic signal. | Erythrocyte staining is part of HPA's reported profile (HPA tissue IHC), while endogenous activity can also produce color in enzyme based detection (general IHC practice). | Check a no primary control and the detection system's endogenous activity control; evaluate glandular cell staining independently (general IHC practice). |
| Kidney tubules stain more weakly than intestinal glands. | That relative pattern is compatible with HPA's Medium score in kidney tubular cells and High score in intestinal glandular cells (HPA tissue IHC). | Score each specified cell population against its own expected level and compare slides processed together before calling the kidney result a failure (general IHC practice). |
Experiment evidence
IHC & IF images showing S1PR2 staining
A02904 · IHCImmunohistochemistry of S1PR2 in mouse brain tissue with S1PR2 antibody at 10 μg/ml.
A02904 · IFImmunofluorescence of S1PR2 in mouse brain tissue with S1PR2 antibody at 20 μg/ml. Red: S1PR2 Antibody (A02904) Blue: DAPI staining
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000267534-S1PR2/tissue/cerebral+cortex#imid_3709392
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000267534-S1PR2/tissue/colon#imid_3709318
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000267534-S1PR2/tissue/liver#imid_3709327
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000267534-S1PR2/tissue/kidney#imid_3709328
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA014255; green = S1PR2, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA014255; green = S1PR2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for S1PR2 IHC & IF
🧪Run duodenum first; its glandular cells should stain (HPA: High in duodenal glandular cells). Use adipose tissue as the negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the duodenum slide, treat morphologically unstained adjacent cells as an internal background reference, without assuming they are S1PR2-negative.
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show S1PR2 in HEK293, SH-SY5Y, U2OS, with annotated localisation: Golgi apparatus (uncertain) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and immunoglobulin class where applicable, and an S1PR2 knockout biological control (standard IHC practice). Block endogenous peroxidase for chromogenic IHC and endogenous biotin if using avidin–biotin detection, particularly in duodenum (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the fixative is unreported in the selected A02904 mouse-brain IHC caption (A02904 caption). The evidence does not establish whether frozen sections or IF are easier: UniProt annotates S1PR2 at the cell membrane, while HPA reports uncertain Golgi and nuclear-speckle ICC-IF localization (UniProt O95136; HPA subcellular). In duodenum, endogenous biotin may confound avidin–biotin detection, so interpret glandular staining against the blocked control (standard IHC practice).
HPA tissue IHC evidence for S1PR2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced S1PR2 IHC Tips
Use matched controls and compartment aware scoring to troubleshoot S1PR2 staining in paraffin sections; assess fluorescence separately.
How should I retrieve S1PR2 when chromogenic staining is weak?
Start with citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval rule). Compare serial sections processed with and without retrieval, keeping antibody concentration and detection conditions constant (standard IHC practice). The selected antibody image shows mouse brain staining at 10 μg/ml, but its caption does not report retrieval or fixation, so it cannot establish which condition produced the image (selected A02904 tissue IHC caption). If staining remains weak, adjust retrieval intensity in small steps and inspect tissue integrity and background before accepting a brighter result (standard IHC practice).
Could fixation explain weak or uneven S1PR2 staining?
Target specific sensitivity to fixation is unknown because the selected tissue IHC caption does not state its fixative (selected A02904 tissue IHC caption). Record fixation history and compare sections prepared under matched conditions before changing antibody concentration; uneven processing can confound chromogenic staining (standard IHC practice). Use the page’s citrate pH 6.0, 95–98 °C, 20 min retrieval consistently while assessing that comparison (page retrieval rule). Check morphology and section edges alongside signal, and interpret any fixation effect as an experimental finding only after matched controls support it (standard IHC practice).
Where should convincing S1PR2 staining appear in tissue?
Prioritise staining compatible with a cell membrane receptor: S1PR2 has 7 transmembrane segments and a UniProt cell membrane annotation (UniProt O95136 topology; UniProt O95136 subcellular location). Examine whether signal outlines plausible cells or a defined membrane surface, then compare that pattern with adjacent tissue and control sections (standard IHC practice). HPA reports distinct staining in erythrocytes and intestinal microvilli, with high glandular cell staining in duodenum and small intestine (HPA tissue IHC). HPA’s mainly Golgi and additional nuclear speckle locations are marked uncertain in ICC/IF, so diffuse nuclear chromogen alone needs independent validation (HPA subcellular; standard IHC practice).
How does epitope position affect S1PR2 staining in paraffin sections?
Map the antibody’s stated immunogen to S1PR2 before interpreting a negative section; the supplied record does not identify an epitope for A02904 (selected A02904 tissue IHC caption; UniProt O95136 topology). S1PR2 has extracellular and cytoplasmic loops, a cytoplasmic C terminus at residues 293–353, and an annotated glycosylation site at residue 19 (UniProt O95136 topology; UniProt O95136 glycosylation). Accessibility can vary with processing and retrieval, so compare matched sections before attributing a difference to epitope masking (standard IHC practice). The record lists 0 isoforms; do not assign divergent staining patterns to a specific isoform without separate evidence (UniProt O95136 isoforms).
How can I cross check chromogenic S1PR2 staining with multiplex IF?
On the separate IF/ICC workflow, pair S1PR2 with a validated marker for the cell population being assessed, using glandular cells in duodenum or small intestine as a tissue context supported by HPA (HPA tissue IHC; standard IF practice). Select spectrally separated fluorophores after checking tissue autofluorescence and include single stain controls to distinguish bleed through from colocalisation (standard IF practice). If the antibody epitope is cytoplasmic, test mild permeabilisation; if extracellular, assess staining without permeabilisation first, since the A02904 caption does not locate its epitope (UniProt O95136 topology; selected A02904 tissue IHC caption; standard IF practice). Compare fluorescence with the chromogenic pattern only in matched tissue regions (standard IHC/IF practice).
What should I check when S1PR2 chromogen appears throughout the section?
Run a no primary control and inspect endogenous peroxidase activity before attributing widespread DAB signal to S1PR2 (standard IHC practice). Check blocking, wash stringency, detection reagent, and chromogen development on adjacent sections, changing 1 variable at a time (standard IHC practice). The selected mouse brain image used 10 μg/ml, but its caption does not establish that concentration for other tissues or processing conditions (selected A02904 tissue IHC caption). Treat erythrocyte or intestinal microvillus staining with particular care: those patterns are reported by HPA, while control sections are still needed to exclude background in the assay at hand (HPA tissue IHC; standard IHC practice).
How should I score S1PR2 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and region of interest before scoring; a membrane associated pattern is biologically plausible for S1PR2 (UniProt O95136 subcellular location; standard IHC practice). Record the percentage of positive cells and intensity on a 0–3 scale, then calculate an H score from 0–300 when intensity differences matter (standard IHC scoring practice). For spatially discrete populations, report positive cells per mm² and normalise to viable tissue area or the number of eligible cells, with the same rule across samples (standard IHC practice). Score erythrocytes and glandular cells separately where present because HPA reports distinct erythrocyte staining and high intestinal glandular cell staining (HPA tissue IHC).
How can I distinguish genuine S1PR2 positivity from artefact?
Seek a reproducible pattern in an anatomically defined cell population that agrees with S1PR2’s membrane annotation and the assay controls (UniProt O95136 subcellular location; standard IHC practice). HPA reports high glandular cell staining in duodenum and small intestine and medium staining in kidney tubules, but describes only medium consistency between staining and RNA expression (HPA tissue IHC). Review unexpected nuclear only signal, section edge enhancement, necrotic areas, and pigment or endogenous enzyme signal against matched controls before calling cells positive (HPA subcellular; standard IHC practice). The mouse brain image documents antibody staining at 10 μg/ml; its unstated fixative and processing do not establish a general positive control standard (selected A02904 tissue IHC caption).
Boster reagents
Best S1PR2 / Sphingosine 1-phosphate receptor 2 IHC Antibodies
A02904 has mouse brain IHC and IF images (catalog image captions). Human, mouse, and rat reactivity is listed for both antibodies (catalog reactivity).
Real IHC data
Anti-S1PR2 Antibody
Cat # A02904
The rendered card is A02904, with IHC-P listed and a mouse brain IHC image at 10 μg/ml (catalog applications; IHC image caption). A02904 also has a mouse brain IF image at 20 μg/ml (IF image caption).
Which to pick: For tissue IHC, choose A02904: IHC-P is listed, and its own image shows mouse brain staining at 10 μg/ml (catalog applications; IHC image caption); the fixative is unreported (IHC image caption). For IF/ICC, A02904-2 lists both applications and is polyclonal, but has no supplied image; A02904 has a mouse brain IF image (catalog applications; dilution details; image captions). Both list human, mouse, and rat reactivity, while the supplied IHC image documents mouse tissue only (catalog reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95136 (S1PR2_HUMAN, Sphingosine 1-phosphate receptor 2).
- Human Protein Atlas. S1PR2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. S1PR2 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the nuclear speckles..
- Human Protein Atlas. S1PR2 antibody validation summary (2 antibodies).
- Critical role of sphingosine-1-phosphate receptor-2 in the disruption of cerebrovascular integrity in experimental stroke. Nature communications 2015 — PMC4587559.
- Characterization of a S1PR2 specific (11)C-labeled radiotracer in streptozotocin-induced diabetic murine model. Nuclear medicine and biology 2023 — PMC10949307.
- Aberrant expression of the S1P regulating enzymes, SPHK1 and SGPL1, contributes to a migratory phenotype in OSCC mediated through S1PR2. Scientific reports 2016 — PMC4861980.
- Synthesis and characterization of [(125)I]TZ6544, a promising radioligand for investigating sphingosine-1-phosphate receptor 2. Nuclear medicine and biology 2020 — PMC8972069.
- PubMed PMID:10617617 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:25274307 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for S1PR2 (Sphingosine 1-phosphate receptor 2). Expected localisation Cell membrane expected (UniProt); microvilli observed (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02904 at 10 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A02904); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-S1PR2, 10 μg/ml (datasheet A02904); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Erythrocyte staining may complicate scoring (HPA tissue IHC); Isoform / epitope: No isoforms annotated; check extracellular vs cytoplasmic epitope location (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt O95136, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A02904, PMC4587559, PMC10949307. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/S1PR2/immunohistochemistry-ihc-guide.