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GNAQ / Guanine nucleotide-binding protein G(q) subunit alpha · IHC design guide
Design Immunohistochemistry for GNAQ
Plan GNAQ paraffin IHC using the variable cytoplasmic and membranous tissue pattern reported by HPA (HPA tissue IHC). Start with the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet: PB9707), and interpret staining cautiously because antibody staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GNAQ 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic and membranous staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9707) | |
| Positive control | Appendix+4 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Low agreement between staining and RNA; possible cross-gene signal (HPA tissue IHC) | |
| Regulation | Reduced in RA peripheral blood lymphocytes (UniProt) | |
| Isoform / epitope | No listed isoforms; one full-length chain (UniProt) |
Section 1
Recommended GNAQ IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet PB9707). The published IHC protocols below cover sheep reproductive tissues, skin, and hen uterovaginal junction (PMC5088417; PMC10392076; PMC8082553).
| Sample | Paraffin-embedded rat ovary tissue; fixative not specified (datasheet PB9707) |
| Fixation | Image fixative and duration unreported (datasheet PB9707); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet PB9707); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9707) |
| Primary antibody | Rabbit anti-GNAQ, 0.5-1μg/ml (datasheet PB9707) |
| Primary incubation | Overnight at 4 °C (datasheet PB9707) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9707) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GNAQ-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and membranous expression at variable levels in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet PB9707); citrate pH 6.0 is documented for the skin and hen protocols (PMC10392076; PMC8082553).
Section 2
What Is the Expected GNAQ Staining Pattern?
GNAQ is a lipid-anchored protein associated with the plasma membrane, with Golgi and nuclear locations also annotated; it has no transmembrane segment (UniProt P50148 topology). In paraffin-section IHC, expect variable cytoplasmic and membranous staining, including in colon and prostate glandular cells (HPA: tissue IHC). Interpret this pattern cautiously: HPA rates the tissue staining Uncertain because antibody staining and RNA expression have low consistency, and the antibody may recognize proteins from more than one gene (HPA: tissue IHC reliability).
| Membranous and cytoplasmic staining in colon or prostate glandular cells, with moderate signal. | This matches the reported compartment pattern and the Medium staining recorded for those cells (HPA: tissue IHC). It is a useful expected result, but HPA's Uncertain reliability means morphology alone does not establish GNAQ specificity (HPA: tissue IHC reliability). |
| Strong, uniform nuclear staining dominates while membranous and cytoplasmic staining is absent. | This departs from the reported tissue IHC pattern (HPA: tissue IHC). Check for detection artefact before scoring it as GNAQ; nuclear and nuclear-membrane locations are annotated by UniProt, so nuclear signal alone is not proof of an artefact (UniProt P50148 subcellular location). |
| Comparable staining appears in glandular cells and nearby cells expected to be unstained. | A pattern spanning unrelated cell types may reflect cross-reactivity or endogenous detection activity (standard IHC practice). HPA reports adipocytes and bone-marrow hematopoietic cells as Not detected, but its IHC reliability is Uncertain; use matched negative controls before assigning identity (HPA: tissue IHC). |
| Pale, diffuse color covers tissue, stroma, and empty areas without clear cell boundaries. | Treat this as background rather than a scored localization pattern (standard IHC practice). Review the no-primary control and detection reagents; genuine tissue staining is described as cytoplasmic and membranous at variable levels (HPA: tissue IHC). |
| No staining is visible in colon or prostate glandular cells on the run. | Both are reported at Medium intensity, so an absent signal calls for a run-level check of antigen retrieval, antibody incubation, detection, and a known-positive control (HPA: tissue IHC; standard IHC practice). A single negative section cannot establish absence of GNAQ. |
💡Expected GNAQ appearanceCall a positive result when identifiable colon or prostate glandular cells show moderate cytoplasmic and membranous staining (HPA: Medium; HPA: tissue IHC pattern); diffuse color without cellular localization or uniform staining across unrelated cells is suspect (standard IHC practice).
| Membrane association and compartment | GNAQ has no transmembrane segment and is associated with the plasma membrane by a lipid anchor (UniProt P50148 topology and subcellular location). Assess membrane-associated signal alongside cytoplasmic staining; HPA reports both in tissue IHC (HPA: tissue IHC). |
| Choice of positive and comparison cells | Colon and prostate glandular cells are Medium, whereas adipocytes and bone-marrow hematopoietic cells are Not detected in HPA tissue IHC (HPA: tissue IHC). Compare defined cell populations within correctly identified sections; these observations are provisional because HPA rates the IHC evidence Uncertain (HPA: reliability). |
| Antibody-specific confidence | HPA lists IHC status as Uncertain for HPA048886 and CAB010036 and warns that the tissue-staining antibody may target protein from more than one gene (HPA: antibodies; HPA: tissue IHC reliability). Confirm an unexpected pattern with an independent specificity control before interpreting it as GNAQ. |
| Processing and isoform expectations | The supplied UniProt record lists one 1–359 chain, no signal peptide or propeptide, and no reported isoforms (UniProt P50148 processing and isoforms). These annotations give no basis for predicting a secreted or shed staining pattern; use the reported cellular locations when interpreting the slide (UniProt P50148 subcellular location). |
| IF/ICC Q: Should the IHC pattern appear identically in IF/ICC? | A: No identical pattern is established. HPA ICC-IF reports supported plasma-membrane localization, with nuclear speckles and cytosol marked uncertain; images are listed for A-431, MCF-7, and U2OS (HPA: subcellular). Interpret IF/ICC on its own guide page; this section provides no IF/ICC protocol. |
| Detection-system background | Endogenous enzyme activity or nonspecific detection can produce color unrelated to primary-antibody binding (standard IHC practice). Judge diffuse signal against a no-primary control and use the detection system's appropriate blocking step when background is present (standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon or prostate glandular cells are blank. | The run may have insufficient detectable signal; both cell populations are reported at Medium staining (HPA: tissue IHC). | Check a known-positive section, then review antigen retrieval, primary-antibody incubation, and detection controls using the IHC-validated antibody's instructions (standard IHC practice). Do not infer a GNAQ-specific fixation effect. |
| All tissue regions develop diffuse chromogen. | Background may arise from endogenous detection activity or nonspecific reagent binding (standard IHC practice). | Inspect the no-primary control, review blocking and washes, and score only cell-associated signal with interpretable morphology (standard IHC practice). |
| Unexpected cells stain as strongly as expected glandular cells. | Cross-reactivity is plausible because HPA cautions that the tissue-staining antibody may target more than one gene (HPA: tissue IHC reliability). | Compare the same cell type in a reported Not detected tissue, where available, and seek independent specificity evidence before calling the unexpected signal GNAQ (HPA: tissue IHC; standard IHC practice). |
| Predominantly nuclear signal appears without membrane or cytoplasmic staining. | It differs from the HPA tissue IHC profile, although UniProt also annotates nuclear locations (HPA: tissue IHC; UniProt P50148 subcellular location). | Review a no-primary control and an expected-positive tissue; withhold a compartment-specific conclusion until the signal is reproducible and antibody specificity is supported (standard IHC practice). |
| Membrane signal is weak but cytoplasmic signal remains visible. | HPA reports variable levels of cytoplasmic and membranous expression; an individual section need not show equal prominence in both compartments (HPA: tissue IHC). | Check cell boundaries and compare the expected-positive control under the same staining conditions before scoring membrane localization (standard IHC practice). |
| A negative comparison tissue shows no staining, but the positive control also fails. | A failed positive control leaves the negative result uninterpretable (standard IHC practice). | Repeat or resolve the IHC run before using absence of color as evidence; HPA's Not detected categories are observational and its tissue IHC reliability is Uncertain (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing GNAQ staining
PB9707 · IHCIHC analysis of GNAQ using anti-GNAQ antibody (PB9707). GNAQ was detected in a paraffin-embedded section of rat ovary tissue. Heat mediated
PB9707 · IHCIHC analysis of GNAQ using anti-GNAQ antibody (PB9707). GNAQ was detected in a paraffin-embedded section of mouse testis tissue. Heat mediat
PB9707 · IFIF analysis of GNAQ using anti-GNAQ antibody (PB9707). GNAQ was detected in immunocytochemical section of A431 cells. Enzyme antigen retriev
M00898 · IHCIHC analysis of GNAQ using anti GNAQ antibody (M00898). GNAQ was detected in paraffin-embedded section of human ovarian cancer tissue. Heat
M00898 · IHCIHC analysis of GNAQ using anti GNAQ antibody (M00898). GNAQ was detected in paraffin-embedded section of human ovarian cancer tissue. Heat
M00898-1 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156052-GNAQ/tissue/cerebral+cortex#imid_13802729
HPA · StomachHuman Protein Atlas IHC, Stomach. Source: https://www.proteinatlas.org/ENSG00000156052-GNAQ/tissue/stomach+1#imid_16239139
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156052-GNAQ/tissue/colon#imid_13803729
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156052-GNAQ/tissue/liver#imid_13803722
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010036; green = GNAQ, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB010036; green = GNAQ, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GNAQ IHC & IF
🧪Run colon first and require staining in its glandular cells (HPA: Colon glandular cells, Medium). Use adipose tissue adipocytes as the negative comparator (HPA: Adipocytes, Not detected); on the colon slide, treat non-glandular cells as an internal background reference only where they are visibly unstained, since their GNAQ status is not specified by the supplied HPA row.
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GNAQ in A-431, MCF-7, U2OS, with annotated localisation: Nuclear speckles (uncertain), Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a matched rabbit IgG isotype control appropriate to the catalog antibody’s clonality (caption: rabbit anti-GNAQ PB9707); use GNAQ-knockout material, if available, as a biological specificity control. For colon, quench endogenous peroxidase and check endogenous biotin background when using the caption’s avidin–biotin/DAB detection system (caption: SABC with DAB).
⚠️Feasibility: The supplied paraffin-section caption uses heat retrieval in EDTA, pH 8.0, followed by 1 μg/ml primary antibody overnight at 4°C; it does not establish that other retrieval conditions fail (caption: PB9707 rat ovary IHC). The fixative is unreported in that caption, and the supplied evidence reports no target-specific fixation window or fixation effect (caption: fixative not stated). HPA has ICC-IF images, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; check colon glands for nonspecific chromogen before scoring (HPA: A-431, MCF-7 and U2OS ICC-IF images; HPA: Colon glandular cells, Medium).
HPA tissue IHC evidence for GNAQ
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Low | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GNAQ IHC Tips
Troubleshoot chromogenic GNAQ IHC in paraffin sections using the catalog antibody’s tissue example, while treating uncertain atlas staining as a validation concern.
Which antigen retrieval should I start with for GNAQ in paraffin sections?
Start with heat-mediated retrieval in EDTA, pH 8.0, for paraffin sections (datasheet PB9707). The selected tissue example used this retrieval before 1 μg/ml primary antibody overnight at 4°C, but its fixative was unreported (datasheet PB9707). Keep section preparation, heating, and cooling consistent across a run so differences in staining can be assessed against matched controls (standard IHC practice). If staining remains weak, vary retrieval duration on adjacent sections while checking tissue morphology and no-primary controls (standard IHC practice). Report the actual heating conditions used, since the caption specifies the buffer and pH but gives no retrieval time (datasheet PB9707).
How should I troubleshoot weak GNAQ staining when fixation details are missing?
Record the fixative, time before fixation, and fixation duration for each specimen before comparing GNAQ staining (standard IHC practice). The selected paraffin-section example does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet PB9707). Compare specimens processed under the same documented conditions, using adjacent sections and the same EDTA, pH 8.0 retrieval (datasheet PB9707; standard IHC practice). If morphology is damaged or staining varies across a block, examine section integrity and processing records before changing antibody concentration (standard IHC practice). The reported 1 μg/ml overnight incubation provides a starting condition, not evidence that any particular fixation condition preserves GNAQ (datasheet PB9707).
Where should credible GNAQ signal appear in chromogenic tissue sections?
First assess membranous and cytoplasmic staining in intact cells, the variable tissue pattern reported for GNAQ IHC (HPA tissue IHC). GNAQ is associated with the plasma membrane through a lipid anchor and has no transmembrane segment (UniProt P50148 topology and subcellular location). Golgi and nuclear membrane locations are annotated, while nuclear speckles and cytosol have uncertain IF support (UniProt P50148 subcellular location; HPA subcellular). Compare staining with cellular boundaries and a counterstain, then check whether the same pattern persists in replicate sections (standard IHC practice). Diffuse staining alone should remain provisional because HPA rates its tissue IHC reliability uncertain and flags possible recognition of protein from another gene (HPA tissue IHC).
Can an unknown epitope explain variable GNAQ staining between specimens?
Check the antibody’s stated immunogen or epitope before assigning a compartment-specific explanation to its IHC pattern (standard IHC practice). The supplied record lists a single 1–359 chain, a G-alpha domain at residues 38–359, and no annotated isoforms (UniProt P50148). A reported modification at residue 209 does not establish that this antibody recognizes that site or that the modification changes staining (UniProt P50148 modified residues; datasheet PB9707). Compare staining across matched sections with the reported EDTA, pH 8.0 retrieval and 1 μg/ml primary condition (datasheet PB9707). If specificity remains uncertain, use an independently validated epitope or biological control before interpreting differences as GNAQ abundance (standard IHC practice; HPA tissue IHC reliability: uncertain).
How can IF help assess an ambiguous chromogenic GNAQ pattern?
Use IF as a complementary localisation check alongside the chromogenic sections, with matched tissue and appropriate fluorescence controls (standard IF/IHC practice). Multiplex GNAQ with an independently validated marker for the cell population being scored, such as glandular cells in colon or prostate sections (HPA tissue IHC: medium glandular-cell staining in colon and prostate; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence in each channel, and include single-label controls to assess bleed-through (standard IF practice). If testing an intracellular epitope, optimise permeabilisation; the antibody’s epitope side is unspecified, and GNAQ has no transmembrane segment (UniProt P50148 topology; datasheet PB9707). Do not transfer the EDTA, pH 8.0 IHC retrieval condition to IF without testing it (datasheet PB9707; standard IF practice).
What should I check when GNAQ DAB staining looks diffuse or widespread?
Run no-primary and secondary-only controls to separate antibody-dependent staining from detection-system background (standard IHC practice). The selected example used 10% goat serum blocking, biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, an avidin-biotin complex, and DAB (datasheet PB9707). Check endogenous peroxidase blocking and, with this detection method, endogenous biotin before interpreting diffuse brown signal (standard chromogenic IHC practice; datasheet PB9707). Keep DAB development consistent across sections and inspect tissue edges, folds, and damaged areas for accentuated staining (standard IHC practice). Even a plausible membranous or cytoplasmic pattern needs controls, because HPA reports uncertain tissue-IHC reliability and possible recognition of protein from another gene (HPA tissue IHC).
How should I score GNAQ staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record percent positive cells and intensity to calculate an H-score (standard IHC practice). For a spatial question, count positive cells per mm² of viable tissue instead, using the same threshold across specimens (standard IHC practice). Normalise counts to viable tissue area or the number of eligible cells, and exclude necrosis, folds, and section edges (standard IHC practice). Score glandular cells separately from surrounding cells when comparing colon or prostate sections, where medium glandular-cell staining is reported (HPA tissue IHC). Use matched retrieval and development conditions, and report uncertainty from HPA’s low staining-to-RNA consistency before equating scores with GNAQ abundance (datasheet PB9707; HPA tissue IHC).
How can I distinguish credible GNAQ positivity from staining artefact?
Give greater weight to reproducible membranous or cytoplasmic staining in intact cells than to signal confined to damaged tissue or section edges (HPA tissue IHC; standard IHC practice). Check whether the stained cell population matches the question: HPA reports medium glandular-cell staining in colon and prostate, but its overall tissue-IHC reliability is uncertain (HPA tissue IHC). Treat isolated nuclear staining cautiously; UniProt lists nuclear locations, whereas HPA marks nuclear speckles uncertain and supports plasma-membrane localisation (UniProt P50148 subcellular location; HPA subcellular). Exclude necrosis, folds, and residual endogenous enzyme signal with morphology and no-primary controls (standard IHC practice). Do not interpret DAB intensity alone as GPCR pathway activation, because GNAQ cycles between GDP- and GTP-bound states (UniProt P50148 function).
Boster reagents
Best GNAQ / Guanine nucleotide-binding protein G(q) subunit alpha IHC Antibodies
The catalog shows GNAQ IHC in paraffin sections from human, mouse and rat tissues, plus IF in A431 cells and an unnamed sample (catalog image captions).
Real IHC data
Anti-GNAQ Antibody ®
Cat # PB9707
Real IHC data
Anti-GNAQ Antibody ® (monoclonal, 13H4)
Cat # M00898
Real IF data
Anti-GNAQ Rabbit Monoclonal Antibody
Cat # M00898-1
PB9707 shows IHC in rat ovary, mouse testis and human prostatic cancer, plus IF in A431 cells (PB9707 image captions). M00898 shows IHC in human ovarian cancer and mouse testis (M00898 image captions); M00898-1 shows IF at 1:50, with no sample identified (M00898-1 IF image caption).
Which to pick: For tissue IHC, choose M00898 for its human ovarian cancer and mouse testis examples, or PB9707 for its rat ovary, mouse testis and human prostatic cancer examples (respective IHC image captions). Both IHC captions describe paraffin sections; the fixative is unreported (respective IHC image captions). For IF/ICC, PB9707 has an A431-cell example at 2 μg/ml (PB9707 IF image caption), while M00898-1 lists IF/ICC and shows IF at 1:50 without identifying the sample (M00898-1 applications and IF image caption). For the broadest stated species reactivity, choose mouse monoclonal M00898, which lists human, monkey, mouse and rat; rabbit PB9707 and rabbit monoclonal M00898-1 list human, mouse and rat (catalog host, clone and reactivity fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50148 (GNAQ_HUMAN, Guanine nucleotide-binding protein G(q) subunit alpha).
- Human Protein Atlas. GNAQ tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. GNAQ subcellular location (ICC-IF): Mainly localized to the nuclear speckles and plasma membrane. In addition localized to the cytosol..
- Human Protein Atlas. GNAQ antibody validation summary (2 antibodies).
- Molecular Cloning, mRNA Expression, and Localization of the G-protein Subunit Galphaq in Sheep Testis and Epididymis. Asian-Australasian journal of animal sciences 2016 — PMC5088417.
- Transcriptome Profiling of Anhidrotic Eccrine Sweat Glands Reveals that Olfactory Receptors on Eccrine Sweat Glands Regulate Perspiration in a Ligand-Dependent Manner. JID innovations : skin science from molecules to population health 2023 — PMC10392076.
- Transcriptomic and metabolomic insights into the variety of sperm storage in oviduct of egg layers. Poultry science 2021 — PMC8082553.
- GNAQ T96S mutation abrogates the ability of wild-type GNAQ to induce apoptosis by phosphorylating annexin A2 in natural killer/T cell lymphoma. Cancer science 2022 — PMC9277252.
- PubMed PMID:8825633 — UniProt-cited evidence.
- PubMed PMID:8664309 — UniProt-cited evidence.
- PubMed PMID:8836152 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GNAQ (Guanine nucleotide-binding protein G(q) subunit alpha). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9707). Validated IHC antibody: PB9707 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat ovary tissue; fixative not specified (datasheet PB9707); 2. Fixation: Image fixative and duration unreported (datasheet PB9707); 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: EDTA pH 8.0 (datasheet PB9707); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9707); 8. Primary antibody: Rabbit anti-GNAQ, 0.5-1μg/ml (datasheet PB9707); 9. Primary incubation: Overnight at 4 °C (datasheet PB9707). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Low agreement between staining and RNA; possible cross-gene signal (HPA tissue IHC); Isoform / epitope: No listed isoforms; one full-length chain (UniProt); Regulation: Reduced in RA peripheral blood lymphocytes (UniProt). Evidence: UniProt P50148, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nuclear speckles and plasma membrane. In…, Datasheet PB9707, PMC5088417, PMC10392076. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GNAQ/immunohistochemistry-ihc-guide.