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GOLGB1 / Golgin subfamily B member 1 · IHC design guide
Design Immunohistochemistry for GOLGB1
Plan GOLGB1 chromogenic IHC scoring around cytoplasmic staining across cells (HPA tissue IHC). Use high staining in adrenal glandular cells and low staining in skeletal myocytes to guide tissue selection (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GOLGB1 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 (HPA tissue IHC); Golgi membrane protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in all cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08226-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Skeletal myocytes may stain weakly (HPA tissue IHC) | |
| Regulation | No expression regulator reported (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage unreported (UniProt) |
Section 1
Recommended GOLGB1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA retrieval (datasheet: EDTA pH 8.0); the published protocol below describes IHC staining of human nucleus pulposus tissue (PMC12887002: IHC methods).
| Sample | Paraffin-embedded human colonic adenocarcinoma tissue; fixative not specified (datasheet A08226-1) |
| Fixation | Image fixative and duration unreported (datasheet A08226-1); 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 A08226-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08226-1) |
| Primary antibody | Rabbit anti-GOLGB1, 1-2 μg/ml (datasheet A08226-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08226-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A08226-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GOLGB1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A08226-1); try compound digestion if adapting the published tissue protocol (PMC12887002: IHC methods).
Section 2
What Is the Expected GOLGB1 Staining Pattern?
GOLGB1 is anchored in the Golgi membrane, with most of the protein facing the cytoplasm (UniProt Q14789 topology). In paraffin-section IHC, expect cytoplasmic staining in many cell types, often concentrated near the nucleus in the Golgi region (HPA: cytoplasmic expression in all cells; HPA: Golgi localization by ICC-IF). HPA rates its tissue staining reliability Enhanced, citing consistency between staining and RNA expression (HPA: tissue IHC reliability).
| A compact, asymmetric cytoplasmic signal near the nucleus appears in glandular or epithelial cells. | This fits Golgi-region staining (HPA: Golgi localization by ICC-IF; UniProt Q14789: Golgi membrane). HPA reports high staining in adrenal gland, appendix, breast, and cervix glandular cells and bronchial respiratory epithelial cells (HPA: tissue IHC). Chromogenic IHC may show a compact patch rather than individual Golgi structures (general IHC practice). |
| The strongest signal fills nuclei, outlines the plasma membrane, or forms a uniform cytoplasmic haze. | These distributions conflict with the expected Golgi-region pattern (HPA: Golgi localization by ICC-IF; UniProt Q14789: Golgi membrane). Treat them as possible artefact or nonspecific staining until control sections and antibody validation resolve the discrepancy (general IHC practice). |
| A cell population stains strongly while expected Golgi-region signal is absent from adjacent cells. | Check cell identity and detection controls before calling cross-reactivity or endogenous detection activity (general IHC practice). GOLGB1 is broadly expressed, so staining outside one highlighted cell type is not automatically wrong (HPA: cytoplasmic expression in all cells; low tissue specificity). The suspect signal is its unexpected distribution or persistence in controls (general IHC practice). |
| Brown signal spreads through tissue, stroma, or empty areas and obscures cell boundaries. | Diffuse staining cannot establish a Golgi-localized positive result (HPA: Golgi localization by ICC-IF). Examine background in the detection controls and review blocking, washes, and chromogen development as general IHC variables (general IHC practice). Do not score broad background as GOLGB1 expression. |
| No convincing signal appears in a section containing an HPA high-staining cell population. | First verify that the expected cells are present and preserved, then inspect assay controls (general IHC practice). HPA reports high staining in, for example, cerebral-cortex neurons, cerebellar granular-layer cells, and colon endothelial cells (HPA: tissue IHC). An absent result in one run does not establish biological absence. |
💡Expected GOLGB1 appearanceCall a result positive when appropriate cells show a distinct, compact Golgi-region cytoplasmic signal, with stronger staining plausible in HPA high populations (HPA: tissue IHC; HPA: Golgi localization by ICC-IF); isolated nuclear staining, uniform haze, or signal that persists in detection controls is suspect (general IHC practice).
| Tissue and cell context | HPA reports cytoplasmic expression across cells and low tissue specificity, alongside high staining in selected populations and low staining in skeletal-muscle myocytes (HPA: tissue IHC). Compare the observed cell population with its stated HPA level; do not use skeletal muscle as an absolute negative control. |
| Golgi membrane topology | GOLGB1 has one transmembrane segment at residues 3236–3256; residues 1–3235 face the cytoplasm (UniProt Q14789 topology). This supports a cytoplasmic Golgi-region expectation, but the supplied record does not map the IHC antibody epitope (UniProt Q14789 topology; HPA: antibody summary). |
| Evidence behind the antibody pattern | HPA lists both HPA011008 and HPA011555 as Enhanced for IHC and ICC (HPA: antibody validation). HPA describes tissue-IHC reliability as consistency between staining and RNA expression (HPA: tissue IHC reliability). These summaries support pattern review; they do not prove that every brown deposit is specific. |
| IF/ICC Q: What localization should I expect? | A: Golgi-apparatus localization is reported as enhanced by HPA ICC-IF (HPA: subcellular). That observation helps interpret an IHC compartment pattern; IF/ICC methods and controls belong in its separate guide. No IF/ICC protocol choice follows from the supplied IHC evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known HPA high-staining population has no visible IHC signal. | The expected cells may be missing from the section, or the assay may have failed (HPA: tissue IHC; general IHC practice). The supplied sources do not establish GOLGB1-specific fixation sensitivity. | Confirm cell identity and section quality, then check a same-run positive control, primary-antibody step, and detection reagents (general IHC practice). Interpret a negative only after the assay performs as expected. |
| Signal is diffuse across tissue or extends into cell-free areas. | Background from blocking, washing, or detection can obscure a compact Golgi-region pattern (general IHC practice; HPA: Golgi localization by ICC-IF). | Compare with an appropriate detection control; review blocking, wash stringency, antibody concentration, and chromogen development according to the assay instructions (general IHC practice). Score only a resolved cellular pattern. |
| Staining is mainly nuclear or forms a continuous cell-surface rim. | That compartment does not match the Golgi membrane assignment (UniProt Q14789: subcellular location; HPA: Golgi localization by ICC-IF). | Check a known staining pattern and control sections, then reassess antibody specificity and detection background before assigning GOLGB1 positivity (general IHC practice). |
| A suspected cell population stains, but the expected cells lack Golgi-region signal. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). Broad GOLGB1 expression means cell identity alone cannot establish an error (HPA: cytoplasmic expression in all cells). | Confirm cell identity, compare compartment patterns, and inspect a primary-antibody omission control for detection activity (general IHC practice). Compare with HPA tissue patterns where the same cell type is documented. |
| Skeletal-muscle myocytes stain faintly beside stronger cells. | HPA records myocytes as low, not negative, and reports low tissue specificity overall (HPA: tissue IHC). Faint cellular signal therefore needs pattern and control review. | Compare myocyte staining with background and the expected Golgi-region distribution; avoid using its low level as proof of assay failure or as an absolute negative control (HPA: tissue IHC; general IHC practice). |
| Golgi-region signal varies across sections or is hard to distinguish from background. | Interpretation may be limited by section quality or assay conditions (general IHC practice). Neither HPA tissue staining nor UniProt topology identifies a GOLGB1-specific fixation or retrieval effect. | Review section integrity, same-run controls, and the validated IHC workflow; document any retrieval or detection changes as general optimization rather than a proven GOLGB1-specific requirement (general IHC practice). |
Experiment evidence
IHC & IF images showing GOLGB1 staining
A08226-1 · IHCIHC analysis of Giantin/GOLGB1 using anti-Giantin/GOLGB1 antibody (A08226-1). Giantin/GOLGB1 was detected in a paraffin-embedded section of
A08226-1 · IHCIHC analysis of Giantin/GOLGB1 using anti-Giantin/GOLGB1 antibody (A08226-1). Giantin/GOLGB1 was detected in a paraffin-embedded section of
A08226-1 · IFIF analysis of Giantin/GOLGB1 using anti-Giantin/GOLGB1 antibody (A08226-1). Giantin/GOLGB1 was detected in an immunocytochemical section of
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000173230-GOLGB1/tissue/cerebral+cortex#imid_3007736
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000173230-GOLGB1/tissue/colon#imid_3012846
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000173230-GOLGB1/tissue/liver#imid_3012855
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000173230-GOLGB1/tissue/kidney#imid_3012840
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA011008; green = GOLGB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA011008; green = GOLGB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GOLGB1 IHC & IF
🧪Run appendix first; its glandular cells should show Golgi-localized staining (HPA: High in appendix glandular cells; UniProt Q14789: Golgi apparatus membrane). HPA detects GOLGB1 in all 45 scored tissues, so use no-primary and isotype controls for the negative comparison; no cell population on the appendix slide is established as an internal negative, although noncellular background should remain unstained (HPA: no negative tissues; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: GOLGB1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GOLGB1 in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Golgi apparatus (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality where known; a GOLGB1 knockout sample, if available, provides a biological specificity control (caption: rabbit primary antibody; standard IHC practice). For the caption’s biotin-based DAB detection, control endogenous peroxidase and biotin signal in the appendix section (caption: streptavidin–biotin complex with DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A08226-1 paraffin-section caption does not state a fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required (caption: heat-mediated antigen retrieval). HPA IF images support assessing Golgi localization in its listed cell lines; the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC, and appendix DAB staining requires attention to endogenous peroxidase and biotin background (HPA: Golgi apparatus localization and ICC-IF images; standard IHC practice).
HPA tissue IHC evidence for GOLGB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: GOLGB1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced GOLGB1 IHC Tips
Use matched paraffin sections and compartment aware scoring to troubleshoot GOLGB1 chromogenic IHC; reserve fluorescence checks for the separate IF/ICC workflow.
How should I adjust retrieval when GOLGB1 staining is weak?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections before GOLGB1 staining (datasheet A08226-1). The selected tissue image used this retrieval, followed by 10% goat serum block and 2 μg/ml primary overnight at 4°C (caption A08226-1). If signal is weak, first check deparaffinisation, incubation and detection, then compare an alternative retrieval condition on serial sections while scoring tissue morphology. Keep retrieval identical across experimental groups, and assess whether staining has a plausible perinuclear Golgi distribution rather than diffuse nuclear colour (UniProt Q14789: Golgi membrane; HPA: Golgi apparatus enhanced).
Could fixation explain weak or uneven GOLGB1 staining?
The selected paraffin-section image does not state a fixative, so target-specific sensitivity to fixative choice, duration or delayed fixation is unknown (caption A08226-1). For a new series, record those variables and compare matched sections using EDTA retrieval at pH 8.0 and the same primary concentration (datasheet A08226-1; caption A08226-1: 2 μg/ml). Examine morphology and perinuclear staining in preserved cells, alongside a no-primary control, before assigning uneven signal to fixation. HPA tissue staining describes GOLGB1 expression and supplies no fixation comparison (HPA: cytoplasmic expression in all cells); assess fixation effects directly in the specimens being studied.
Where should convincing GOLGB1 chromogenic staining appear?
Expect a compact, often perinuclear cytoplasmic distribution consistent with the Golgi apparatus, allowing for its shape in sectioned cells (UniProt Q14789: Golgi membrane; HPA: Golgi apparatus enhanced). GOLGB1 has a cytoplasmic region spanning residues 1–3235 and a membrane anchor at 3236–3256, so widespread nuclear DAB is biologically discordant (UniProt Q14789 topology). HPA reports cytoplasmic tissue expression broadly, including high staining in colon endothelial cells; inspect cell identity before comparing compartments across fields (HPA: cytoplasmic expression in all cells; High in colon endothelial cells). Use a no-primary section and well-preserved areas to judge whether diffuse colour reflects detection background or tissue damage.
Can isoforms or epitope position change the IHC result?
GOLGB1 has 4 annotated isoforms, but the supplied antibody evidence does not identify its epitope or establish isoform selectivity (UniProt Q14789: isoforms 1–4; caption A08226-1). Its cytoplasmic sequence extends through residue 3235, followed by a membrane segment at 3236–3256 and a short lumenal tail (UniProt Q14789 topology). Document the antibody's immunogen before claiming recognition of a particular isoform or membrane side; compare staining across matched sections under identical retrieval and detection conditions. Multiple modified residues, including phosphoserines at 6 and 17, are annotated, but their effects on this antibody's tissue staining are untested (UniProt Q14789 modified residues).
How can IF help assess a questionable IHC pattern?
For a separate IF/ICC check, look for GOLGB1 signal at the Golgi and multiplex with an epithelial cell marker when assessing glandular cells (HPA: Golgi apparatus enhanced; High in appendix glandular cells). Choose a fluorophore in a channel with low specimen autofluorescence, often far red, and include single-colour and no-primary controls to assess bleed-through and background. Set permeabilisation according to the verified antibody epitope: GOLGB1 residues 1–3235 face the cytoplasm, whereas residues 3257–3259 face the lumen (UniProt Q14789 topology). The epitope is unspecified here, so test permeabilisation empirically and do not transfer a fluorescence result directly into a chromogenic IHC score.
What should I check when DAB background obscures GOLGB1?
Check a no-primary section first, then inspect whether colour follows tissue edges, damaged regions or the perinuclear pattern expected for GOLGB1 (UniProt Q14789: Golgi membrane; HPA: Golgi apparatus enhanced). The selected method used 10% goat serum, biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, and a streptavidin-biotin complex with DAB (caption A08226-1). In that workflow, assess endogenous peroxidase and biotin background with appropriate controls, and apply a peroxidase block as a general chromogenic IHC step. If background persists, compare shorter primary exposure or stronger washing on serial sections while holding retrieval at EDTA pH 8.0 (datasheet A08226-1).
How should I quantify GOLGB1 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring; HPA reports cytoplasmic staining across cells and high staining in colon endothelial cells (HPA: cytoplasmic expression in all cells; High in colon endothelial cells). Score an H-score from intensity categories 0–3 and their percentages, or report the percentage of positive cells with an explicit perinuclear pattern rule. For spatial analyses, report positive cell density per mm² of evaluable tissue and normalise measurements to viable tissue area or the number of cells in the predefined population. Keep section thickness, retrieval at EDTA pH 8.0, staining batch and image thresholds consistent across specimens (datasheet A08226-1).
How do I distinguish true GOLGB1 staining from artefact?
Treat a defined cytoplasmic Golgi pattern in intact cells as more plausible than nuclear or uniformly diffuse DAB, given GOLGB1's membrane association and enhanced Golgi localisation (UniProt Q14789: Golgi membrane; HPA: Golgi apparatus enhanced). Check cell identity against the sampled tissue: HPA reports high staining in colon endothelial cells and low staining in skeletal muscle myocytes (HPA: High in colon endothelial cells; Low in skeletal muscle myocytes). Exclude section edges and necrotic or poorly preserved areas from interpretation, and inspect a no-primary control for endogenous enzyme or detection colour. Confirm questionable fields on adjacent sections using the same EDTA pH 8.0 retrieval before assigning biological meaning (datasheet A08226-1).
Boster reagents
Best GOLGB1 / Golgin subfamily B member 1 IHC Antibodies
A08226-1 has real IHC data from human paraffin sections and IF data from RT4 cells (catalog IHC and IF image captions; catalog: Human reactivity).
Real IHC data
Anti-Giantin/GOLGB1 Antibody ®
Cat # A08226-1
A08226-1 is listed for human IHC, with paraffin-section images from colonic adenocarcinoma, esophageal squamous carcinoma, gall bladder adenosquamous carcinoma, and placenta (catalog applications and reactivity; A08226-1 IHC image captions). The same SKU is listed for IF/ICC and has an IF image from RT4 cells (catalog applications; A08226-1 IF image caption).
Which to pick: Choose A08226-1 for human tissue IHC on paraffin sections; its own captions document EDTA pH 8 retrieval and 2 μg/ml primary antibody (A08226-1 IHC image captions). For IF/ICC, A08226-1 lists both applications and shows IF in RT4 cells at 5 μg/ml (catalog applications; A08226-1 IF image caption). Its listed reactivity is Human, so the payload supports no cross-species recommendation; the IHC captions do not report a fixative (catalog: Human reactivity; A08226-1 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14789 (GOGB1_HUMAN, Golgin subfamily B member 1).
- Human Protein Atlas. GOLGB1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GOLGB1 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. GOLGB1 antibody validation summary (2 antibodies).
- GOLGB1 deficiency accelerates intervertebral disc degeneration by activating the MAPK pathway. Scientific reports 2026 — PMC12887002.
- PubMed PMID:8198703 — UniProt-cited evidence.
- PubMed PMID:7802676 — UniProt-cited evidence.
- PubMed PMID:18487259 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GOLGB1 (Golgin subfamily B member 1). Expected localisation Cytoplasmic staining (HPA tissue IHC); Golgi membrane protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08226-1). Validated IHC antibody: A08226-1 at 1-2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colonic adenocarcinoma tissue; fixative not specified (datasheet A08226-1); 2. Fixation: Image fixative and duration unreported (datasheet A08226-1); 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 A08226-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08226-1); 8. Primary antibody: Rabbit anti-GOLGB1, 1-2 μg/ml (datasheet A08226-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08226-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Skeletal myocytes may stain weakly (HPA tissue IHC); Isoform / epitope: 4 isoforms; epitope coverage unreported (UniProt); Regulation: No expression regulator reported (UniProt). Evidence: UniProt Q14789, HPA tissue IHC · Enhanced, HPA ICC-IF · Golgi apparatus., Datasheet A08226-1, PMC12887002. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GOLGB1/immunohistochemistry-ihc-guide.