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- Table of Contents
Plan chromogenic GOLPH3 IHC in paraffin sections using the IHC-validated antibody and a Golgi-like cytoplasmic readout (datasheet A01333-1; HPA tissue IHC). Use stomach glandular cells as a high-staining reference and adipocytes as an undetected-cell comparator when scoring by cell type and intensity (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Golgi-like cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Golgi-like cytoplasm in glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01333-1) | |
| Positive control | Stomach+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); effects are uncertain. | |
| Caveat | Staining varies despite low RNA tissue specificity (HPA tissue IHC) | |
| Regulation | Detected in mitochondrial-disease muscle (UniProt) | |
| Isoform / epitope | No isoforms annotated; one 1–298 chain; epitope unmapped (UniProt) |
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A01333-1). The four published GOLPH3 IHC protocols below provide tissue-specific conditions (PMC4007226; PMC3480894; PMC5624675; PMC5356558).
| Sample | Paraffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01333-1) |
| Fixation | Image fixative and duration unreported (datasheet A01333-1); 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 A01333-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01333-1) |
| Primary antibody | Rabbit anti-GOLPH3, 2-5μg/ml (datasheet A01333-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01333-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01333-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GOLPH3-positive staining in glandular cells of stomach (HPA tissue IHC: High). HPA tissue profile: Golgi-like cytoplasmic expression in several tissues. No signal in the no-primary control. |
GOLPH3 should produce Golgi-like cytoplasmic staining in several tissues, especially stomach glandular cells (HPA: tissue IHC, High). It associates with Golgi membranes without a transmembrane segment (UniProt Q9H4A6: localization and topology). Interpret a paraffin-section result against the tissue and cell type on the slide: HPA rates its tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Concentrated Golgi-like cytoplasmic stain in stomach glandular cells, stronger than surrounding unstained areas (HPA: stomach glandular cells, High). | This fits the reported tissue pattern and Golgi localization (HPA: tissue IHC; UniProt Q9H4A6: localization). Judge the compartment within identifiable glandular cells rather than calling any brown material positive (standard IHC practice). |
| Predominantly nuclear stain, with little or no Golgi-like cytoplasmic signal. | A dominant nuclear pattern conflicts with the reported Golgi-like tissue profile (HPA: tissue IHC). Treat it as a possible staining artefact and review controls; UniProt also lists endosome, cell membrane, and mitochondrial locations, so a non-Golgi cytoplasmic signal needs context (UniProt Q9H4A6: localization). |
| Strong stain in adipocytes or esophageal squamous epithelial cells (HPA: both cell types, Not detected). | This disagrees with the sampled HPA tissue observations and raises possible cross-reactivity or endogenous detection activity (HPA: tissue IHC; standard IHC practice). It is a control question, not proof that every cell of that type must always be negative (HPA: medium consistency). |
| Diffuse brown haze across cells, stroma, and empty slide areas, without a distinct Golgi-like pattern. | This does not match the reported Golgi-like cytoplasmic profile (HPA: tissue IHC). Broad background can obscure a genuine compartmental signal; compare a no-primary control and review blocking, washes, and detection conditions (standard IHC practice). |
| No convincing signal in an adequately sampled stomach glandular-cell area (HPA: stomach glandular cells, High). | The expected positive tissue failed to show its reported pattern (HPA: tissue IHC). Check tissue preservation and the IHC run with appropriate controls, then review retrieval and antibody conditions; this result alone does not establish absent GOLPH3 (standard IHC practice). |
| Where should the signal sit? | Golgi stack and trans-Golgi network membranes are annotated, with recruitment involving phosphatidylinositol 4-phosphate binding and oligomerization (UniProt Q9H4A6). Assess the predominant cellular pattern as Golgi-like cytoplasmic staining (HPA: tissue IHC). |
| Which cells help interpret intensity? | Stomach glandular cells are High; glandular cells in colon, appendix, breast, cervix, and endometrium are Medium (HPA: tissue IHC). Use the named cell type when comparing fields: HPA also records Not detected adipocytes and esophageal squamous epithelial cells (HPA: tissue IHC). |
| How firm is the tissue pattern? | The tissue IHC assessment is Approved with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability). HPA044564 and HPA055841 are each IHC Approved; that status should not be described as IHC Enhanced (HPA: antibody validation). |
| Does topology predict a surface stain? | GOLPH3 has no transmembrane segment, signal peptide, or propeptide, and one annotated chain spanning residues 1–298 (UniProt Q9H4A6: topology and processing). Although cell membrane is an annotated location, those features do not make a uniform plasma-membrane outline the expected tissue IHC pattern (UniProt Q9H4A6; HPA: tissue IHC). |
| What about IF/ICC? | On its separate guide page, IF/ICC can be compared with HPA's supported Golgi-apparatus and vesicle localization; HPA055841 has Supported ICC validation (HPA: subcellular; antibody validation). That cell-image evidence does not assign vesicular staining to a specific paraffin-section cell type (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Stomach glandular cells show no stain (HPA: High). | The section or run may not have yielded an interpretable positive result; a blank slide cannot distinguish failed detection from biology (standard IHC practice). | Confirm glandular cells are present, inspect the positive and no-primary controls, and review the catalog antibody's IHC-P retrieval and detection conditions (standard IHC practice). |
| Only nuclear staining is prominent. | Nuclear dominance disagrees with the Golgi-like cytoplasmic tissue profile (HPA: tissue IHC) and suggests nonspecific signal or detection artefact (standard IHC practice). | Compare the no-primary control, reduce avoidable background through blocking and washes, and reassess whether any cytoplasmic Golgi-like signal remains (standard IHC practice). |
| Adipocytes or esophageal squamous epithelial cells stain strongly (HPA: Not detected). | The result conflicts with the sampled HPA observations; cross-reactivity or endogenous detection activity is possible, but the mismatch alone does not identify its cause (HPA: tissue IHC; standard IHC practice). | Check a no-primary control and the same-run positive tissue, then assess the suspect cells separately from nearby positive structures (standard IHC practice). |
| Brown color spreads through tissue without cell boundaries or a Golgi-like focus. | Diffuse background can hide the reported Golgi-like cytoplasmic pattern (HPA: tissue IHC; standard IHC practice). | Inspect the no-primary control and review blocking, washing, chromogen development, and counterstain so cellular localization can be judged (standard IHC practice). |
| Colon glandular staining is weaker than stomach glandular staining. | That difference can agree with HPA's Medium colon and High stomach observations; intensity alone does not show a failed run (HPA: tissue IHC). | Score each cell type and compartment against its own reported level, with same-run controls; avoid imposing stomach-level intensity on colon (HPA: tissue IHC; standard IHC practice). |
| Puncta appear outside the compact Golgi-like area. | UniProt annotates endosome and other locations, while HPA IF/ICC supports vesicles; those records do not establish that every punctum in tissue IHC is specific (UniProt Q9H4A6; HPA: subcellular). | Check whether the dominant glandular-cell pattern remains Golgi-like, compare controls, and record extra puncta separately rather than scoring them as the defining IHC result (HPA: tissue IHC; standard IHC practice). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot GOLPH3 staining by checking retrieval, compartment pattern, tissue controls, and scoring before interpreting changes in chromogenic signal.
Anti-GOLPH3 antibodies have IHC images from human intestinal cancer and mouse and rat brain paraffin sections, plus IF images from A431 and rat lung cells (catalog image captions).
A01333-1 lists IHC and IF/ICC and shows paraffin-section IHC in human intestinal cancer, mouse brain, and rat brain, plus IF in A431 cells (catalog applications; catalog image captions). A01333 lists IF and shows staining in rat lung cells (catalog applications; catalog IF caption).
Which to pick: Choose A01333-1 for tissue IHC: it lists IHC for human, mouse, and rat and has paraffin-section images in all three; the fixative is unreported (catalog applications/reactivity; catalog IHC captions). For IF/ICC, A01333-1 lists both applications and shows A431-cell IF, while A01333 lists IF and shows rat lung-cell IF (catalog applications; catalog IF captions). A01333-1 has direct cross-species IHC image evidence; both antibodies are rabbit antibodies with no clone reported (catalog IHC captions; catalog host/clone).