GOLPH3 / Golgi phosphoprotein 3 · IHC design guide

Design Immunohistochemistry for GOLPH3

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).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for GOLPH3 (IHC for GOLPH3): expected localisation Golgi-like cytoplasmic staining (HPA tissue IHC), antibody A01333-1, validated IHC image, and IHC protocol steps
Printable GOLPH3 IHC protocol sheet — expected localisation Golgi-like cytoplasmic staining (HPA tissue IHC), antibody A01333-1, controls and protocol steps. Open the full GOLPH3 IHC guide →

GOLPH3 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended GOLPH3 IHC & IF Protocols

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).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human intestinal cancer tissue; fixative not specified (datasheet A01333-1)
FixationImage fixative and duration unreported (datasheet A01333-1); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A01333-1); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A01333-1)
Primary antibodyRabbit anti-GOLPH3, 2-5μg/ml (datasheet A01333-1)
Primary incubationOvernight at 4 °C (datasheet A01333-1)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet A01333-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultGOLPH3-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.
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A01333-1); use an article’s retrieval conditions when reproducing its published method (cited PMC).
Section 2

What Is the Expected GOLPH3 Staining Pattern?

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).

What am I looking at on my slide?
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).
💡Expected GOLPH3 appearanceCall positive a concentrated Golgi-like cytoplasmic signal in glandular cells, typically High in stomach and Medium in colon (HPA: tissue IHC); dominant nuclear stain or slide-wide brown haze is suspect (HPA: Golgi-like profile; standard IHC practice).
How each factor affects the staining
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).
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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).

Sample controls for GOLPH3 IHC & IF

🧪Run stomach first: glandular cells should stain (HPA: High in stomach glandular cells). Use adipose tissue as the biological negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the stomach slide, compare glandular staining with surrounding nonglandular cells as a background reference, without assuming those cells are GOLPH3-negative.
Positive control tissue: Stomach (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GOLPH3 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Golgi apparatus (supported), Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where known, and a GOLPH3 knockout sample if available (caption: rabbit primary and anti-rabbit secondary; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection and assess any background in the stomach section against the no-primary slide (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: The selected A01333-1 paraffin-section caption does not report a fixative, and the supplied evidence reports no target-specific fixation window or fixation effect. Heat retrieval in EDTA at pH 8.0 was used successfully in that caption, but retrieval dependence has not been established (A01333-1 tissue-IHC caption). HPA documents Golgi and vesicle staining by ICC-IF, but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC; interpret diffuse glandular signal cautiously against the expected Golgi-associated pattern (HPA: supported Golgi apparatus and vesicle localization; UniProt Q9H4A6: Golgi membrane localization).

HPA tissue IHC evidence for GOLPH3

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced GOLPH3 IHC Tips

Troubleshoot GOLPH3 staining by checking retrieval, compartment pattern, tissue controls, and scoring before interpreting changes in chromogenic signal.

Which retrieval conditions should I start with for weak GOLPH3 staining?
Start with heat-mediated retrieval in EDTA pH 8.0 for this paraffin-section assay (datasheet A01333-1). The selected tissue image used that retrieval before incubation with 2 μg/ml antibody overnight at 4°C, so keep those conditions consistent while assessing retrieval (datasheet A01333-1). Compare a weak section with a concurrently processed positive control, recording heating time and tissue integrity (standard IHC practice; HPA: high staining in stomach glandular cells). If staining remains weak, test a limited retrieval-time series on adjacent sections; excessive heating can damage morphology and make the expected Golgi-like cytoplasmic pattern harder to judge (standard IHC practice; HPA: Golgi-like cytoplasmic expression).
Could fixation explain loss of GOLPH3 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A01333-1). Record fixative, fixation duration, tissue thickness, and processing history before comparing cases, because differences in preparation can alter antigen accessibility and morphology (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the antibody at 2 μg/ml while evaluating preparation effects on adjacent sections (datasheet A01333-1). Include a consistently processed positive control and compare its punctate, Golgi-like cytoplasmic staining with the test section; poor control staining supports a processing or assay problem, without identifying fixation as its cause (standard IHC practice; HPA: Golgi-like cytoplasmic expression).
What staining pattern should count as GOLPH3 in chromogenic IHC?
Look for a compact, Golgi-like cytoplasmic signal associated with cells rather than uniform nuclear staining (HPA: Golgi-like cytoplasmic expression; UniProt Q9H4A6: Golgi membranes). GOLPH3 is also annotated at vesicles and other compartments, so a broader cytoplasmic pattern requires control-based evaluation rather than automatic rejection (HPA subcellular: Golgi apparatus and vesicles; UniProt Q9H4A6: endosome, cell membrane, mitochondrial intermembrane space). Its recruitment to Golgi membranes involves phosphatidylinositol 4-phosphate binding and oligomerization, and it has no transmembrane segment (UniProt Q9H4A6: localisation and topology). Compare staining with adjacent negative-control sections at the same magnification, checking whether the apparent pattern persists within intact cells (standard IHC practice).
How should I assess epitope-dependent differences between GOLPH3 antibodies?
The supplied record lists a single 1–298 chain and 0 annotated isoforms, so an isoform-specific explanation is unsupported here (UniProt Q9H4A6: processing and isoforms). It lists phosphoserines at positions 35 and 36, but no antibody epitope is provided; do not attribute staining differences to those modifications without epitope mapping or direct testing (UniProt Q9H4A6: modified residues; datasheet A01333-1: epitope not stated). Compare antibodies on adjacent sections using their individually validated retrieval conditions and matched detection controls (standard IHC practice). Concordant Golgi-like cytoplasmic staining provides support, while disagreement calls for independent specificity controls before assigning a biological explanation (HPA: Golgi-like cytoplasmic expression; standard IHC practice).
How can IF help resolve an ambiguous chromogenic GOLPH3 pattern?
Use IF/ICC as a separate assay to check whether GOLPH3 overlaps a Golgi marker and appears in the expected cell population; HPA reports supported Golgi and vesicle localisation (HPA subcellular: Golgi apparatus and vesicles). In multiplex tissue IF, pair it with a marker identifying the cells being scored, and choose spectrally separated fluorophores after checking tissue autofluorescence in an unstained control (standard IF practice). Optimise permeabilisation empirically for antibody access to the cytosolic face of Golgi membranes: GOLPH3 has no transmembrane segment and associates with Golgi membranes through lipid binding (UniProt Q9H4A6: topology and recruitment; standard IF practice). Do not carry over the paraffin-section retrieval or fixation assumptions to IF/ICC (datasheet A01333-1: paraffin IHC caption only).
What should I check when DAB obscures the Golgi-like signal?
Inspect a no-primary control for staining from the detection system, including endogenous peroxidase activity, before changing the primary antibody concentration (standard IHC practice). Apply an appropriate peroxidase block, wash thoroughly, and shorten DAB development if diffuse precipitate is masking cellular detail (standard IHC practice). The selected tissue image used 10% goat serum block, 2 μg/ml primary overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (datasheet A01333-1). If background persists, titrate the primary and compare intact tissue with section edges and damaged areas; accept staining only when cellular morphology supports a Golgi-like cytoplasmic pattern (standard IHC practice; HPA: Golgi-like cytoplasmic expression).
How should I score GOLPH3 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring region before reviewing case labels, then score only intact cells with interpretable cytoplasm (standard IHC practice). Record the percentage of positive cells and intensity grades, or calculate an H-score from those grades and percentages; keep the same threshold and illumination across sections (standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Report glandular and other cell populations separately where relevant, because HPA records differing GOLPH3 staining levels among tissue cell types (HPA: tissue IHC profile and cell-specific staining levels).
When is apparent GOLPH3 positivity more likely to be artefact?
Favor staining that resolves into Golgi-like cytoplasmic signal within intact cells and appears in a concurrently stained positive control (HPA: Golgi-like cytoplasmic expression; standard IHC practice). Treat isolated nuclear signal cautiously because the supported subcellular pattern is Golgi and vesicular (HPA subcellular: Golgi apparatus and vesicles). Check cell identity: stomach glandular cells show high staining, whereas adipocytes are listed as not detected, although neither observation alone validates an individual section (HPA: stomach glandular cells high; adipocytes not detected). Discount staining confined to section edges or necrotic areas, and compare no-primary controls to identify endogenous peroxidase or detection background before assigning a biological change (standard IHC practice).
Boster reagents

Best GOLPH3 / Golgi phosphoprotein 3 IHC Antibodies

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).

Real IHC data IHC analysis of GOLPH3 using anti-GOLPH3 antibody (A01333-1). GOLPH3 was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-GOLPH3 Antibody (A01333-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-GOLPH3 Antibody ®
Cat # A01333-1
Real IF data Immunofluorescence of GOLPH3 in Rat Lung cells with GOLPH3 antibody at 20 μg/mL.
Anti-Golgi phosphoprotein 3 GOLPH3 Antibody
Cat # A01333

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).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry Q9H4A6 (GOLP3_HUMAN, Golgi phosphoprotein 3).
  2. Human Protein Atlas. GOLPH3 tissue IHC expression (reliability: Approved).
  3. Human Protein Atlas. GOLPH3 subcellular location (ICC-IF): Localized to the Golgi apparatus and vesicles..
  4. Human Protein Atlas. GOLPH3 antibody validation summary (2 antibodies).
  5. GOLPH3 is a novel marker of poor prognosis and a potential therapeutic target in human renal cell carcinoma. British journal of cancer 2014 — PMC4007226.
  6. GOLPH3 overexpression correlates with tumor progression and poor prognosis in patients with clinically N0 oral tongue cancer. Journal of translational medicine 2012 — PMC3480894.
  7. GOLPH3 and YB-1 Are Novel Markers Correlating With Poor Prognosis in Prostate Cancer. World journal of oncology 2015 — PMC5624675.
  8. GOLPH3 overexpression correlates with poor response to neoadjuvant therapy and prognosis in locally advanced rectal cancer. Oncotarget 2016 — PMC5356558.
  9. PubMed PMID:11042173 — UniProt-cited evidence.
  10. PubMed PMID:14702039 — UniProt-cited evidence.
  11. PubMed PMID:16710414 — UniProt-cited evidence.