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GPR132 / Probable G protein-coupled receptor 132 · IHC design guide
Design Immunohistochemistry for GPR132
Plan chromogenic GPR132 IHC on paraffin sections using the catalog antibody's 1:100–1:300 dilution range (datasheet A07182-2). Assess cytoplasmic staining across immune and other cells (HPA tissue IHC), and interpret membrane or endosomal localisation as a molecular expectation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GPR132 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 | Cytoplasmic staining in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining, including immune cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07182-2) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Hippocampus+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07182-2); assess its effects empirically. | |
| Caveat | Staining and RNA expression show low consistency (HPA tissue IHC) | |
| Regulation | Expression is enriched in macrophages (UniProt) | |
| Isoform / epitope | 3 isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt) |
Section 1
Recommended GPR132 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A07182-2) with a published GPR132 IHC protocol for human breast tissue microarrays (PMC5047746).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07182-2) |
| Fixation | Image fixative and duration unreported (datasheet A07182-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A07182-2); 20 min, 95–100 °C (standard) |
| 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-GPR132, 1:100-1:300 (datasheet A07182-2) |
| 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 | GPR132-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression, including immune cells. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER (datasheet A07182-2). The published protocol heated an unspecified retrieval buffer at 95°C for 1 hour (PMC5047746).
Section 2
What Is the Expected GPR132 Staining Pattern?
GPR132 is a seven-transmembrane receptor annotated at the cell membrane, with internalization into endosomal compartments; macrophages and lymphocyte-rich hematopoietic tissues are expected sites (UniProt Q9UNW8). HPA reports high staining in bone marrow hematopoietic cells and a broader cytoplasmic tissue pattern (HPA: tissue IHC). Its IHC reliability is Supported, although staining has low consistency with RNA data and awaits external verification (HPA: reliability description).
| Membrane-associated staining in hematopoietic cells, with some intracellular puncta. | This fits the receptor's membrane location and possible endosomal accumulation (UniProt Q9UNW8). High bone marrow hematopoietic-cell staining is reported (HPA: tissue IHC). Puncta alone cannot identify endosomes in chromogenic IHC; assess the full cellular pattern and controls. |
| Predominantly nuclear staining or staining confined to extracellular material. | Neither location matches the annotated membrane and endosomal distribution (UniProt Q9UNW8). Treat an isolated pattern there as suspect; check morphology, detection controls and whether the signal remains after changing detection conditions. |
| Strong staining in an unexpected cell population, especially when controls also develop color. | Consider antibody cross-reactivity or endogenous detection activity as general IHC possibilities. HPA also reports high staining in several epithelial and glandular populations (HPA: tissue IHC), so cell identity alone cannot establish a false positive. |
| Diffuse color across tissue, empty spaces and the section edge. | A pattern that does not follow cells is more consistent with background than receptor localization (general IHC practice). Compare a no-primary control, inspect tissue edges, and judge any cellular signal against that background. |
| No staining in bone marrow hematopoietic cells. | This conflicts with the reported high staining for those cells (HPA: bone marrow tissue IHC), but one negative section does not establish absence of GPR132. Check tissue preservation, retrieval, antibody conditions and detection with appropriate controls (general IHC practice). |
💡Expected GPR132 appearanceA credible positive shows cell-associated membrane and/or intracellular staining in hematopoietic cells, potentially strong in bone marrow (UniProt Q9UNW8; HPA: High in bone marrow hematopoietic cells); nuclear-only color or diffuse color outside cells is suspect (UniProt Q9UNW8; general IHC practice).
| Compartment and trafficking | GPR132 is annotated at the cell membrane and can accumulate in endosomal compartments; LPC-driven movement to the surface is annotated by similarity (UniProt Q9UNW8). A cytoplasmic appearance therefore needs interpretation alongside membrane staining, without assuming that every cytoplasmic signal is specific. |
| Cell population and tissue context | Macrophages and lymphocyte-rich hematopoietic tissues are highlighted by UniProt (UniProt Q9UNW8). HPA reports high staining in bone marrow hematopoietic cells and several epithelial or glandular cell groups (HPA: tissue IHC). Score identified cells within each tissue rather than assigning one expected intensity to the entire section. |
| Reference-pattern uncertainty | HPA calls IHC reliability Supported but reports low agreement between antibody staining and RNA expression, pending external verification (HPA: reliability description). Its ubiquitous cytoplasmic profile should be treated as an observed staining pattern, not proof that every cytoplasmic signal represents GPR132. |
| Topological epitope access | The receptor has seven transmembrane segments and extracellular and cytoplasmic regions (UniProt Q9UNW8 topology). Antibody epitopes are not supplied here, so these data cannot specify an antigen-retrieval condition or predict which compartment the catalog antibody will emphasize. |
| Isoforms and glycosylation | Three isoforms and one glycosylation site at residue 35 are annotated (UniProt Q9UNW8). Without an antibody epitope or isoform-specific validation, these annotations cannot explain a particular IHC intensity difference or establish isoform-specific staining. |
| Reference negatives | Glial cells in hippocampus and red-pulp cells in spleen are listed as not detected (HPA: tissue IHC). These are cell-specific observations; the spleen entry does not contradict UniProt's broader statement about expression in lymphocyte-rich spleen tissue (UniProt Q9UNW8). |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow hematopoietic cells remain unstained. | The result differs from the reported high IHC signal in that cell population (HPA: bone marrow tissue IHC); section or assay failure is possible (general IHC practice). | Verify the intended cells and section quality, then review the antibody's IHC-P instructions, retrieval, dilution and detection controls. Reassess before calling the tissue negative (general IHC practice). |
| Color is predominantly nuclear. | Nuclear localization is unsupported by the membrane and endosomal annotation (UniProt Q9UNW8); nonspecific binding or detection background is possible (general IHC practice). | Compare a no-primary control, check counterstain and chromogen separation, and evaluate whether any cell-associated membrane or cytoplasmic pattern persists (general IHC practice). |
| Most of the section has uniform brown haze. | Uniform haze can reflect background from antibody concentration, blocking or detection chemistry (general IHC practice); HPA's broad cytoplasmic profile does not validate acellular haze (HPA: tissue IHC). | Review no-primary and reagent controls, blocking and washes; adjust antibody conditions according to its IHC-P instructions, then compare cellular signal with background (general IHC practice). |
| Unexpected epithelial or glandular cells stain strongly. | Several such cell groups are reported as high by HPA (HPA: tissue IHC), while low staining–RNA agreement limits confidence in assigning every positive cell to GPR132 (HPA: reliability description). | Identify the stained cell type and compare its reported tissue pattern. Use appropriate negative controls and independent specificity evidence before labeling the population cross-reactive (general IHC practice). |
| Spleen red-pulp cells show little or no signal. | Red-pulp cells are listed as not detected by HPA (HPA: spleen tissue IHC). UniProt's spleen statement concerns lymphocyte-rich hematopoietic tissue and does not specify red-pulp cells (UniProt Q9UNW8). | Score red pulp separately from other spleen compartments; avoid using absent red-pulp staining alone to reject an otherwise controlled run (HPA: spleen tissue IHC; general IHC practice). |
| Intracellular staining is strong but membrane staining is hard to resolve. | Endosomal accumulation is annotated by UniProt, while HPA describes broad cytoplasmic IHC staining (UniProt Q9UNW8; HPA: tissue IHC). Chromogenic resolution and background can obscure a thin membrane rim (general IHC practice). | Check cellular boundaries and background at suitable magnification, compare no-primary controls, and report the observed compartment without claiming endosomal identity from IHC alone (general IHC practice). |
Experiment evidence
IHC & IF images showing GPR132 staining
A07182-2 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use
A30804 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200(4° overnight). 2, Tris-EDTA,pH9.0 was used
A07182 · IFImmunofluorescence analysis of A549. 1, primary Antibody was diluted at 1:200 (4°C overnight). 2, Goat Anti Rabbit IgG (H&L) - Alexa Fluor 4
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000183484-GPR132/tissue/cerebral+cortex#imid_7812012
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000183484-GPR132/tissue/colon#imid_7811954
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000183484-GPR132/tissue/liver#imid_7811939
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000183484-GPR132/tissue/kidney#imid_7811940
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GPR132 IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). Use hippocampus as the negative tissue because glial cells are listed as not detected (HPA: Not detected in hippocampal glial cells); on the positive slide, assess morphologically distinct nonhematopoietic cells as internal negatives only after confirming their identity and staining status.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Hippocampus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GPR132; derive a cell-line control from the positive tissue's cell type (Hematopoietic cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and a GPR132 knockout control or peptide block where a matching immunogen peptide is available (standard IHC control practice). Quench endogenous peroxidase in bone marrow sections before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected A07182-2 paraffin-section caption also leaves the fixative unreported (selected-SKU tissue-IHC caption). That caption uses Tris-EDTA at pH 9.0 for antigen retrieval and a 1:200 primary dilution overnight at 4°C, but does not establish whether retrieval is required under other conditions (selected-SKU tissue-IHC caption). Whether frozen sections or IF are easier is unreported; membrane and endosomal staining are plausible localisation patterns (UniProt Q9UNW8 subcellular annotation), while marrow’s dense hematopoietic cell population calls for careful cell-level scoring (HPA: High in bone marrow hematopoietic cells).
HPA tissue IHC evidence for GPR132
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced GPR132 IHC Tips
Troubleshoot GPR132 staining in paraffin sections using the catalog antibody’s documented retrieval conditions, cellular context, and controls.
Which retrieval conditions should I start with for GPR132 paraffin sections?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 (datasheet A07182-2). The documented paraffin-section image used this buffer before overnight primary incubation at 4°C with antibody diluted 1:200 (caption A07182-2). If staining is weak, compare a milder retrieval condition on adjacent sections while keeping detection and exposure to chromogen constant (standard IHC practice). Check tissue preservation and the presence of interpretable cells before increasing retrieval intensity, since excessive heating can damage section morphology (standard IHC practice). Record the retrieval temperature and duration for each run so differences can be traced to processing rather than assigned to GPR132 abundance (standard IHC practice).
How should I handle fixation when GPR132 staining is weak?
Target-specific sensitivity of GPR132 staining to fixation is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (caption A07182-2). Record the actual fixative, fixation duration, and processing history for each specimen before comparing staining across cases (standard IHC practice). Use similarly processed control and test sections, then apply the documented Tris-EDTA pH 9.0 retrieval and 1:200 primary dilution consistently (datasheet A07182-2; caption A07182-2). If signal differs between processing batches, repeat with matched sections and inspect morphology before changing antibody concentration (standard IHC practice). Do not assign a GPR132-specific fixation mechanism without direct comparison (supplied evidence: no fixation study).
Should GPR132 staining appear at the membrane or inside cells?
Evaluate both membrane-associated and intracellular patterns: GPR132 is annotated at the cell membrane and can accumulate in endosomal compartments (UniProt Q9UNW8 subcellular location). Its 7 transmembrane segments support a membrane-associated interpretation, while the reported endosomal pool makes exclusively surface staining an unsuitable requirement (UniProt Q9UNW8 topology and subcellular location). The tissue IHC profile describes ubiquitous cytoplasmic expression, including immune cells, but reports low consistency between staining and RNA data (HPA tissue IHC). Score membrane and cytoplasmic staining separately in intact cells, excluding folds and detached tissue (standard IHC practice). Compare each compartment against negative-control staining before calling diffuse chromogen deposition positive (standard IHC practice).
Can this IHC antibody distinguish GPR132 isoforms or membrane faces?
Do not assign an isoform from this stain alone: GPR132 has 3 annotated isoforms, and the supplied antibody caption gives no epitope sequence (UniProt Q9UNW8 isoforms; caption A07182-2). The receptor has extracellular segments at residues 1–45 and several loops, plus a cytoplasmic tail at 312–380 (UniProt Q9UNW8 topology). An N-linked glycosylation site is annotated at residue 35, but its effect on this antibody’s tissue staining is untested here (UniProt Q9UNW8 glycosylation; supplied evidence). Obtain epitope mapping and isoform sequence coverage before interpreting differential staining as isoform-specific (standard IHC validation practice). Keep retrieval and detection identical when comparing sections (standard IHC practice).
How would I check this IHC pattern with multiplex immunofluorescence?
On the separate IF/ICC workflow, pair GPR132 staining with a validated marker of the cell population under study, such as a macrophage marker when testing the reported macrophage enrichment (UniProt Q9UNW8 tissue specificity; standard IF practice). Select spectrally separated fluorophores and inspect an unstained section for tissue autofluorescence before assigning a weak channel as positive (standard IF practice). Permeabilisation should follow the mapped epitope: intracellular loops or the cytoplasmic tail require access through the membrane, whereas an extracellular epitope may be assessed without that step (UniProt Q9UNW8 topology; standard IF practice). The supplied paraffin IHC caption provides no IF fixation or antibody conditions (caption A07182-2). Validate IF signal with appropriate single-label and negative controls (standard IF practice).
What should I check when GPR132 chromogen looks widespread?
Compare the tissue with a no-primary control to identify secondary-reagent binding and endogenous enzyme signal (standard IHC practice). Include a peroxidase block before DAB development and assess endogenous pigment or precipitate separately from cell-bound chromogen (standard chromogenic IHC practice). Broad cytoplasmic staining has been reported, including in immune cells, but the tissue IHC profile has low consistency with RNA expression (HPA tissue IHC). Titrate around the documented 1:200 primary dilution while holding retrieval at Tris-EDTA pH 9.0 and development time constant (caption A07182-2; standard IHC practice). Reject staining concentrated at section edges, folds, or damaged areas when scoring cells (standard IHC practice).
How should I quantify GPR132 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because GPR132 has reported membrane and endosomal localisation while tissue IHC describes cytoplasmic staining (UniProt Q9UNW8 subcellular location; HPA tissue IHC). Within each annotated region, report the percentage of positive intact cells and an intensity-weighted H-score, or positive-cell density per mm² when cell counts vary (standard IHC quantification practice). Normalise positive-cell counts to the number of eligible cells, and area-based counts to measured viable tissue area (standard IHC quantification practice). Keep retrieval, 1:200 primary dilution, chromogen development, and scoring thresholds consistent across compared sections (caption A07182-2; standard IHC practice). Report membrane and cytoplasmic results separately (standard IHC quantification practice).
How can I distinguish plausible GPR132 staining from artefact?
Prioritise staining in intact cells within a defined compartment; GPR132 is membrane-associated and can occupy endosomal compartments (UniProt Q9UNW8 subcellular location). Macrophages and lymphocyte-rich hematopoietic tissues are reported expression contexts, while the HPA tissue IHC profile also reports strong staining in several epithelial and glandular cell populations (UniProt Q9UNW8 tissue specificity; HPA tissue IHC). Treat that breadth cautiously because HPA reports low consistency between antibody staining and RNA expression (HPA tissue IHC reliability). Compare candidate signal with no-primary controls, adjacent morphology, and any independently validated cell markers before assigning cell identity (standard IHC practice). Exclude edge staining, necrotic regions, pigment, and endogenous peroxidase signal from a positive call (standard IHC practice).
Boster reagents
Best GPR132 / Probable G protein-coupled receptor 132 IHC Antibodies
Real IHC images show paraffin-embedded human tonsil for A07182-2 and A30804 (catalog IHC captions); a real IF image shows A549 for A07182 (catalog IF caption).
Real IHC data
Anti-GPR132 Antibody
Cat # A07182-2
Real IHC data
Anti-GPR132/G2A Antibody
Cat # A30804
Real IF data
Anti-G2A GPR132 Antibody
Cat # A07182
A07182-2 and A30804 each have an IHC image of paraffin-embedded human tonsil, with primary antibody at 1:200 after Tris-EDTA pH 9.0 retrieval (respective IHC captions). A07182 has an IF image of A549 with primary antibody at 1:200 and a DAPI counterstain (A07182 IF caption).
Which to pick: For tissue IHC, choose A07182-2 or A30804: each lists IHC and shows staining in paraffin-embedded human tonsil; the fixative is unreported in both captions (respective application lists and IHC captions). For IF, A07182 has an A549 image; for ICC, A30804 lists ICC as an application (A07182 IF caption; A30804 application list). For species coverage, A30804 lists human, mouse and rat, while A07182 lists human, monkey and mouse; these are listed reactivities, not image evidence for every species (respective catalog reactivity lists).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UNW8 (GP132_HUMAN, Probable G protein-coupled receptor 132).
- Human Protein Atlas. GPR132 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. GPR132 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. GPR132 antibody validation summary (2 antibodies).
- Expression of pH-Sensitive GPCRs in Peritoneal Carcinomatosis of Colorectal Cancer-First Results. Journal of clinical medicine 2023 — PMC10003041.
- Expression profiles of proton-sensing G-protein coupled receptors in common skin tumors. Scientific reports 2020 — PMC7501253.
- Macrophage PPARγ inhibits Gpr132 to mediate the anti-tumor effects of rosiglitazone. eLife 2016 — PMC5047746.
- Lactate activates CCL18 expression via H3K18 lactylation in macrophages to promote tumorigenesis of ovarian cancer. Acta biochimica et biophysica Sinica 2024 — PMC11543520.
- PubMed PMID:9770487 — UniProt-cited evidence.
- PubMed PMID:19855098 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GPR132 (Probable G protein-coupled receptor 132). Expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A07182-2). Validated IHC antibody: A07182-2 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Hippocampus. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A07182-2); 2. Fixation: Image fixative and duration unreported (datasheet A07182-2); 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 retrieval: Tris-EDTA pH 9.0 (datasheet A07182-2); 20 min, 95–100 °C (standard); 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-GPR132, 1:100-1:300 (datasheet A07182-2); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07182-2); assess its effects empirically.; Caveat: Staining and RNA expression show low consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt); Regulation: Expression is enriched in macrophages (UniProt). Evidence: UniProt Q9UNW8, HPA tissue IHC · Supported, HPA ICC-IF · Membrane, Datasheet A07182-2, PMC5047746. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GPR132/immunohistochemistry-ihc-guide.