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GPR158 / Metabotropic glycine receptor · IHC design guide
Design Immunohistochemistry for GPR158
Plan GPR158 paraffin-section IHC using a catalog antibody at 1:100–1:300 and a peptide-blocked control (datasheet: A11699, A30819 IHC). Interpret nuclear and cytoplasmic CNS staining cautiously because tissue-IHC reliability is uncertain (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GPR158 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 | Nuclear and cytoplasmic staining in CNS (HPA tissue IHC) | |
| Staining pattern | CNS cells show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Cerebral cortex+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 paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining; reliability uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No isoforms annotated; extracellular vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended GPR158 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with one published chromogenic IHC protocol for mouse and human prostate sections (PMC4333349).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A11699) |
| Fixation | Image fixative and duration unreported (datasheet A11699); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-GPR158, 1:100-1:300 (datasheet A11699) |
| 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 | GPR158-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression in CNS. No signal in the no-primary control. |
💡Decision noteStart with the catalog protocol’s Tris-EDTA pH 9.0 retrieval (page retrieval); citrate pH 6.0 is the published prostate alternative (PMC4333349).
Section 2
What Is the Expected GPR158 Staining Pattern?
GPR158 is a seven-pass receptor mainly assigned to postsynaptic membranes, with a smaller presynaptic pool and reported nuclear localisation (UniProt Q5T848). In tissue IHC, expect staining in CNS neuronal cells: high in cerebral cortex and medium in hippocampus and the cerebellar granular layer (HPA tissue IHC). HPA describes nuclear and cytoplasmic CNS staining, but rates its tissue IHC evidence Uncertain because staining and RNA data have low consistency and presumed off-target binding was disregarded (HPA tissue IHC).
| Neuronal staining in cerebral cortex, with weaker staining in hippocampus or the cerebellar granular layer. | This follows HPA's reported high cortical and medium hippocampal and granular-layer staining (HPA tissue IHC). Interpret membrane-associated signal in light of GPR158's predominantly postsynaptic localisation (UniProt Q5T848); assess nuclear or cytoplasmic signal cautiously because HPA reports both while rating the IHC evidence Uncertain (HPA tissue IHC). |
| Predominantly nuclear signal with little neuronal membrane-associated staining. | A nuclear pool is reported, and HPA describes nuclear CNS staining (UniProt Q5T848; HPA tissue IHC). Nuclear-only staining does not establish specificity: the receptor is mainly assigned to the postsynaptic membrane, and HPA's tissue IHC reliability is Uncertain (UniProt Q5T848; HPA tissue IHC). Compare controls before calling it artefact. |
| Strong signal in adipocytes, glandular cells or hematopoietic cells while CNS neurons are weak. | HPA reports no detection in adipocytes of adipose tissue, glandular cells of adrenal gland or appendix, and bone marrow hematopoietic cells (HPA tissue IHC). Such a reversal raises concern for nonspecific antibody binding or endogenous chromogenic activity; check the staining distribution and detection controls (general IHC practice). |
| Uniform colour across cells, extracellular spaces and slide background. | This distribution cannot be read as receptor localisation without cellular boundaries (general IHC practice). Uneven reagent coverage, excess detection background or retained chromogen can produce diffuse colour (general IHC practice). Compare a no-primary control and inspect tissue morphology before assigning any positive cells (general IHC practice). |
| No detectable signal in cerebral cortex neurons. | Cortex is HPA's strongest listed tissue observation, at high staining in neuronal cells (HPA tissue IHC). A blank result calls for checking section integrity, retrieval, antibody conditions and detection performance (general IHC practice). It does not by itself establish absent GPR158, particularly given HPA's Uncertain IHC reliability (HPA tissue IHC). |
💡Expected GPR158 appearanceA plausible positive is high staining in cerebral cortex neurons, with membrane-associated signal consistent with the mainly postsynaptic assignment; HPA also reports nuclear and cytoplasmic CNS signal, while broad staining of HPA-listed negative cells is suspect (HPA tissue IHC; UniProt Q5T848).
| Compartment and topology | GPR158 has 7 transmembrane segments, an extracellular region at residues 24–417 and a cytoplasmic tail at 665–1215 (UniProt Q5T848 topology). Interpret a membrane-associated pattern against this architecture; the observed stain also depends on which region the antibody recognises, which is unspecified here. |
| Protein processing and glycosylation | UniProt lists a signal peptide at residues 1–23, a chain beginning at residue 24, and five glycosylation sites in the extracellular region (UniProt Q5T848). These annotations identify regions to consider when an antibody epitope is known; they do not establish shedding, retrieval needs or fixation sensitivity. |
| Tissue choice | Cerebral cortex neurons provide the strongest listed IHC-positive comparison; hippocampal neurons and cerebellar granular-layer cells are listed at medium staining (HPA tissue IHC). HPA lists several cell types as not detected, including adipose adipocytes and bone marrow hematopoietic cells (HPA tissue IHC). Compare like cell types when scoring. |
| Strength of validation | HPA rates tissue IHC Uncertain, citing low agreement between antibody staining and RNA expression and presumed off-target binding that was disregarded (HPA tissue IHC). Its listed antibody HPA013185 is also IHC Uncertain (HPA antibodies). Treat apparent positives as provisional until their distribution and controls agree. |
| Detection background | Endogenous enzyme activity and nonspecific detection can create chromogenic colour unrelated to antigen (general IHC practice). A no-primary control helps separate detection background from primary-dependent staining (general IHC practice). No supplied source establishes a GPR158-specific fixation effect or an optimal retrieval condition. |
| Situation | Likely cause | Next action |
|---|---|---|
| Cortex is blank. | A high neuronal signal is reported for cerebral cortex, but HPA rates the tissue IHC evidence Uncertain (HPA tissue IHC). Technical failure remains possible (general IHC practice). | Check morphology and section coverage, then verify retrieval, antibody incubation and chromogenic detection with appropriate run controls (general IHC practice). Record the conditions before interpreting the blank as biological absence. |
| Negative comparison cells stain strongly. | Adipose adipocytes and bone marrow hematopoietic cells are listed as not detected (HPA tissue IHC). Primary binding or detection background may explain unexpected colour (general IHC practice). | Compare a no-primary control and review the cell boundaries and signal distribution (general IHC practice). Avoid scoring colour outside the expected cells as GPR158 without additional specificity evidence. |
| Staining is diffuse or obscures cell outlines. | Excess detection background or uneven reagent coverage can mask cellular localisation (general IHC practice). HPA's Uncertain rating increases the need to inspect the pattern critically (HPA tissue IHC). | Inspect the no-primary control, assess washing and chromogen development, and rescore only where morphology remains clear (general IHC practice). |
| Only nuclei are positive. | UniProt reports nuclear localisation but describes GPR158 as mainly postsynaptic; HPA reports nuclear CNS staining despite Uncertain tissue IHC reliability (UniProt Q5T848; HPA tissue IHC). | Compare the same cell population with membrane-associated and cytoplasmic staining, plus controls (general IHC practice). Report nuclear-only staining as a qualified observation rather than a confirmed receptor distribution. |
| Hippocampus is weaker than cortex. | This ordering matches HPA's medium neuronal staining in hippocampus versus high neuronal staining in cerebral cortex (HPA tissue IHC). | Score intensity by cell population and retain the tissue identity in the record (general IHC practice). Do not require hippocampal neurons to match cortical intensity. |
| IF/ICC: should membrane fluorescence be expected? | HPA's ICC-IF summary says membrane, but supplies no cell-line images; its listed antibody has no ICC validation entry (HPA subcellular; HPA antibodies). | Use the membrane assignment as a localisation hypothesis, not an IF validation claim (HPA subcellular; UniProt Q5T848). Consult the separate IF/ICC guide for assay design. |
Experiment evidence
IHC & IF images showing GPR158 staining
A11699 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using GPR158 Antibody. The picture on the right is blocked with the s
A11699 · IFImmunofluorescence analysis of HUVEC cells, using GPR158 Antibody. The picture on the right is blocked with the synthesized peptide.
A30819 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using GPR158 Antibody. The picture on the right is blocked with the s
A30819 · IFImmunofluorescence analysis of HUVEC cells, using GPR158 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151025-GPR158/tissue/cerebral+cortex#imid_3520296
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151025-GPR158/tissue/colon#imid_3520230
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151025-GPR158/tissue/liver#imid_3520239
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151025-GPR158/tissue/kidney#imid_3520224
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GPR158 IHC & IF
🧪Run cerebral cortex first and expect neuronal cells to stain (HPA: High in cerebral cortex neuronal cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the cortex slide, use non-neuronal cells as an internal background reference, while confirming their staining empirically.
Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GPR158; derive a cell-line control from the positive tissue's cell type (Neuronal cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class (standard IHC practice); use a GPR158 knockout or a peptide-blocked primary as a biological specificity control (selected A11699 tissue-IHC caption: peptide-blocked image). Block endogenous peroxidase for chromogenic detection, and assess brain autofluorescence if interpreting IF (standard IHC/IF practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the fixative for the paraffin-section brain image is unreported (selected A11699 tissue-IHC caption: fixative not stated). Retrieval dependence is also unreported, so evaluate antigen retrieval empirically for the IHC-validated antibody (standard IHC practice). The evidence does not establish whether frozen sections or IF are easier; brain autofluorescence can complicate IF interpretation (standard IF practice), and no ICC-IF cell-line images are listed (HPA: subcellular record).
HPA tissue IHC evidence for GPR158
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | 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 GPR158 IHC Tips
Troubleshoot GPR158 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
How should I retrieve GPR158 in paraffin sections when staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval setting). Cool sections consistently before blocking and compare a cerebral cortex control across runs, because neuronal staining there is reported as high but the tissue IHC assessment is uncertain (HPA: cerebral cortex high; HPA: reliability uncertain). If signal remains weak, vary heating time in a small pilot while keeping antibody and detection conditions fixed (standard IHC practice). Record nuclear, cytoplasmic and membrane-associated staining separately, since the reported CNS tissue pattern and the annotated membrane location differ (HPA: CNS profile; UniProt Q5T848 localisation).
Could fixation explain variable GPR158 staining between paraffin blocks?
GPR158-specific fixation sensitivity is unknown from the supplied evidence; the catalog image describes paraffin-embedded human brain tissue but does not state a fixative (A11699 caption). For new specimens, use a consistent fixation and processing schedule, and document fixative, duration and section age for each block (standard IHC practice). If existing blocks stain differently, compare them in the same pH 9.0 retrieval and chromogenic run before attributing the difference to fixation (page retrieval setting; standard IHC practice). Include a same-run brain control and assess tissue preservation, because variable processing can complicate interpretation of staining intensity (HPA: CNS profile; standard IHC practice).
Where should convincing GPR158 chromogenic signal appear?
Assess membrane-associated staining in neuronal profiles, while recording nuclear and cytoplasmic signal separately rather than merging compartments into one score (UniProt Q5T848 localisation; HPA: CNS profile). GPR158 is annotated mainly at the postsynaptic membrane, with a smaller presynaptic portion, and nuclear trafficking is reported with an unresolved mechanistic question (UniProt Q5T848 localisation). Human tissue IHC reports nuclear and cytoplasmic expression in the CNS, but its reliability is uncertain because staining and RNA data show low consistency and presumed off-target binding (HPA: CNS profile; HPA: reliability uncertain). Compare patterns with a peptide-blocked section where feasible; the selected brain image includes that comparison, which alone cannot establish every stained compartment as specific (A11699 caption; standard IHC interpretation).
How does an unknown antibody epitope affect GPR158 staining?
Confirm the antibody's immunogen or mapped epitope before interpreting compartment-specific signal; the supplied caption does not identify its sequence (A11699 caption). The record lists 7 transmembrane segments, a large extracellular region at residues 24–417 and a cytoplasmic tail at 665–1215, so epitope position matters when assessing accessibility (UniProt Q5T848 topology; standard IHC practice). It also lists 5 glycosylation sites and several phosphoserines, but supplies no evidence that these modifications change staining with this antibody (UniProt Q5T848 modifications). No isoforms are annotated here; do not assign different nuclear and membrane patterns to distinct isoforms without independent evidence (UniProt Q5T848 isoforms; standard IHC interpretation).
How can I check GPR158 localisation by multiplex immunofluorescence?
Pair GPR158 with a neuronal marker and inspect overlap by compartment, since neuronal cells are reported positive in cerebral cortex and hippocampus (HPA: cerebral cortex high; HPA: hippocampus medium). Choose spectrally separated fluorophores, favoring a longer-wavelength channel for a weak target when tissue autofluorescence is prominent, and include single-stain controls (standard IF practice). If the antibody epitope is cytoplasmic, pilot mild detergent permeabilisation; if extracellular, compare with no detergent, because the annotated receptor has domains on both sides of the membrane (UniProt Q5T848 topology; standard IF practice). Treat this as an IF optimisation experiment: the supplied tissue image documents paraffin-section IHC, and the subcellular payload lists no ICC/IF images (A11699 caption; HPA: subcellular payload).
What should I change when GPR158 DAB signal is diffuse or widespread?
Run a no-primary control through peroxidase blocking and DAB development to check detection background in the same tissue (standard chromogenic IHC practice). Compare antibody and detection reagent titrations while holding the pH 9.0 retrieval run constant; excessive reagent or development can obscure compartment boundaries (page retrieval setting; standard IHC practice). Check whether staining tracks section edges, folds or damaged regions, and compare with an intact area on that slide (standard IHC practice). Widespread signal in cell types reported as undetected, such as adipocytes in adipose tissue, warrants scrutiny rather than automatic acceptance, especially given the uncertain tissue IHC reliability (HPA: adipose tissue not detected; HPA: reliability uncertain).
How should I quantify GPR158 staining across brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and cell classes before scoring, then report the percentage of positive neuronal cells and an intensity-weighted H-score on a fixed 0–300 scale (standard IHC scoring practice). Score nuclear, cytoplasmic and membrane-associated signal separately because the tissue profile includes the first two while the protein record emphasizes the membrane (HPA: CNS profile; UniProt Q5T848 localisation). For area-based analysis, report positive cells per mm² and normalise to the counted neuronal population or intact tissue area, stating which denominator was used (standard IHC quantification practice). Apply identical retrieval, imaging thresholds and exclusion rules across groups, and present the uncertain HPA reliability alongside biological comparisons (page retrieval setting; standard IHC practice; HPA: reliability uncertain).
Which findings support a genuine GPR158-positive result rather than artefact?
Look for reproducible neuronal staining in intact cerebral cortex or hippocampus, with compartment scores reported separately (HPA: cerebral cortex high; HPA: hippocampus medium; standard IHC practice). Membrane-associated signal is consistent with the receptor annotation, whereas nuclear and cytoplasmic signal also occurs in the tissue IHC profile and needs cautious interpretation (UniProt Q5T848 localisation; HPA: CNS profile; HPA: reliability uncertain). Reject patterns confined to edges, necrotic tissue or folds, and investigate DAB signal in the no-primary control as possible endogenous enzyme or detection background (standard chromogenic IHC practice). Peptide blocking in the selected brain image is supporting evidence, but the uncertain tissue IHC assessment means a blocked image alone does not settle specificity (A11699 caption; HPA: reliability uncertain).
Boster reagents
Best GPR158 / Metabotropic glycine receptor IHC Antibodies
Both antibodies show paraffin-embedded human brain IHC with peptide-blocking images (catalog IHC captions) and HUVEC IF images (catalog IF captions); both list Human and Mouse reactivity (catalog reactivity).
Real IHC data
Anti-GPR158 Antibody
Cat # A11699
Real IHC data
Anti-GPR158 Antibody
Cat # A30819
A11699 shows paraffin-embedded human brain IHC with peptide blocking and HUVEC IF (A11699 image captions); its listed applications include IHC and IF (A11699 applications). A30819 shows the same sample types in its own images (A30819 image captions) and also lists ICC (A30819 applications).
Which to pick: For tissue IHC, either SKU has its own paraffin-embedded human brain image with peptide blocking (A11699 IHC caption; A30819 IHC caption); the fixative is unreported in both captions. For IF, both have HUVEC images (A11699 IF caption; A30819 IF caption); choose A30819 when ICC is required because only it lists ICC (A30819 applications; A11699 applications). For Mouse samples, both list Mouse reactivity and are rabbit polyclonals (catalog reactivity and dilution data), but the supplied IHC images show human brain (A11699 IHC caption; A30819 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5T848 (MGLYR_HUMAN, Metabotropic glycine receptor).
- Human Protein Atlas. GPR158 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. GPR158 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. GPR158 antibody validation summary (1 antibodies).
- Inhibition of GPR158 by microRNA-449a suppresses neural lineage of glioma stem/progenitor cells and correlates with higher glioma grades. Oncogene 2018 — PMC6072706.
- Orphan receptor GPR158 controls stress-induced depression. eLife 2018 — PMC5823542.
- Expression and functional role of orphan receptor GPR158 in prostate cancer growth and progression. PloS one 2015 — PMC4333349.
- Trilobatin, a Naturally Occurring GPR158 Ligand, Alleviates Depressive-like Behavior by Promoting Mitophagy. Journal of agricultural and food chemistry 2025 — PMC11887424.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:10574461 — UniProt-cited evidence.
- PubMed PMID:12168954 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GPR158 (Metabotropic glycine receptor). Expected localisation Nuclear and cytoplasmic staining in CNS (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A11699 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A11699); 2. Fixation: Image fixative and duration unreported (datasheet A11699); 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 Tris-EDTA buffer, pH 9.0 , 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-GPR158, 1:100-1:300 (datasheet A11699); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining; reliability uncertain (HPA tissue IHC); Isoform / epitope: No isoforms annotated; extracellular vs cytoplasmic epitope matters (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q5T848, HPA tissue IHC · Uncertain, HPA ICC-IF · Membrane, Datasheet A11699, PMC4333349. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GPR158/immunohistochemistry-ihc-guide.