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GBF1 / Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 · IHC design guide
Design Immunohistochemistry for GBF1
Plan GBF1 paraffin-section IHC using the catalog antibody’s 2–5 μg/ml range (datasheet A01817-1) and the granular cytoplasmic tissue pattern (HPA tissue IHC). Use cell-specific staining to choose controls; Golgi association is a molecular expectation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GBF1 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 | Granular cytoplasm in several tissues (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm in glandular and neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01817-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 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 A01817-1) | |
| Caveat | Tissue staining has only medium RNA concordance (HPA tissue IHC) | |
| Regulation | Ubiquitous expression (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended GBF1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A01817-1) with the published human brain paraffin-section preparation (PMC13084705 methods).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A01817-1) |
| Fixation | Image fixative and duration unreported (datasheet A01817-1); 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 A01817-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01817-1) |
| Primary antibody | Rabbit anti-GBF1, 2-5 μg/ml (datasheet A01817-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01817-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01817-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GBF1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Granular cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A01817-1); sodium citrate at 95°C for 30 minutes is a published alternative (PMC13084705 methods).
Section 2
What Is the Expected GBF1 Staining Pattern?
GBF1 is a cytoplasmic protein associated with the Golgi, including the cis-Golgi network, and the ER–Golgi intermediate compartment; it has no transmembrane segment (UniProt Q92538). In paraffin-section IHC, expect granular cytoplasmic staining in several cell types, including glandular, respiratory epithelial, and neuronal cells reported at medium intensity (HPA tissue IHC). Treat this pattern as provisional because HPA rates tissue-IHC reliability Uncertain (HPA tissue IHC).
| Granular cytoplasmic staining in colon glandular cells, bronchial respiratory epithelium, or cerebellar Purkinje cells. | These are examples of medium staining reported by HPA (HPA tissue IHC). A compact perinuclear component is consistent with Golgi association (UniProt Q92538; HPA subcellular ICC-IF), but chromogenic IHC alone cannot identify the stained organelle. Score cell type, intensity, and distribution together. |
| Predominantly nuclear staining, or staining confined to cell surfaces or extracellular material. | That distribution does not match the reported granular cytoplasmic tissue pattern or supported Golgi location (HPA tissue IHC; HPA subcellular ICC-IF). Consider nonspecific binding, detection background, or a scoring error; morphology alone cannot determine which explanation applies. |
| Strong staining in adipocytes or duodenal glandular cells while the expected positive cells are weak. | HPA reports GBF1 as not detected in those cells, so disproportionate signal raises concern for cross-reactivity or detection background (HPA tissue IHC). It is not proof of a false positive: the tissue-IHC assessment is Uncertain, and absence of HPA staining does not establish biological absence (HPA tissue IHC). |
| Diffuse color covers most cells or tissue structures without a discernible granular cytoplasmic pattern. | The distribution is difficult to interpret as GBF1 because the reported tissue pattern is granular cytoplasmic (HPA tissue IHC). Review a no-primary control and the detection chemistry using general IHC practice; background, excessive detection signal, and poor contrast can obscure compartment and cell-type boundaries. |
| No signal in the chosen positive tissue, including the cells expected to stain. | First confirm that the section contains the relevant cell type: HPA reports medium staining in colon glandular cells, among other examples (HPA tissue IHC). An absent result can reflect assay performance or sampling; HPA's Uncertain tissue-IHC rating prevents treating one negative section as definitive evidence that GBF1 is absent (HPA tissue IHC). |
💡Expected GBF1 appearanceA plausible positive is medium, granular cytoplasmic staining in an HPA-reported cell type such as colon glandular cells, with possible perinuclear enrichment consistent with Golgi association; isolated nuclear, surface, or extracellular color is suspect (HPA tissue IHC; HPA subcellular ICC-IF; UniProt Q92538).
| Compartment and dynamics | GBF1 cycles rapidly on and off early Golgi membranes and is also listed in cytoplasm and the ER–Golgi intermediate compartment (UniProt Q92538). Expect a cytoplasmic pattern that may include Golgi-region enrichment, without requiring every positive cell to show an identical compact spot. |
| Tissue and cell selection | HPA reports medium staining in several glandular, respiratory epithelial, and neuronal populations, but not detected staining in selected other cells (HPA tissue IHC). Select and score a documented cell population rather than assigning one expected intensity to an entire tissue. |
| Validation limit | The tissue-IHC profile has Uncertain reliability; both listed HPA antibodies have Uncertain IHC assessments, while HPA037759 has Supported ICC assessment (HPA tissue IHC; HPA antibodies). These ratings support cautious interpretation and do not establish IHC specificity for a different catalog antibody. |
| Topology and processing | GBF1 has no transmembrane segment or signal peptide, and UniProt lists a single chain spanning residues 1–1860 (UniProt Q92538). A membrane outline or extracellular deposit is therefore not the expected primary pattern; these annotations do not establish how a particular epitope behaves in paraffin sections. |
| Isoforms and modified residues | UniProt lists four isoforms and multiple phosphorylated residues (UniProt Q92538). Their effect on staining cannot be predicted without the antibody epitope and validation data; do not interpret intensity differences as isoform expression or phosphorylation from this IHC pattern alone. |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells show no chromogenic signal. | The relevant cells may be absent from the section, or the staining run may have failed; HPA's reported positives are cell-specific (HPA tissue IHC). | Check the section for an HPA-reported medium cell population, such as colon glandular cells, and compare a concurrently stained positive control (HPA tissue IHC; general IHC practice). Review the catalog antibody's IHC-P instructions for the actual retrieval, dilution, and detection settings. |
| Color is widespread and obscures cellular detail. | The reported GBF1 tissue pattern is granular cytoplasmic, so diffuse color may reflect assay background (HPA tissue IHC). | Compare with a no-primary control; assess blocking, washing, and detection development as general IHC practice. If an enzyme-based detection system is used, check its relevant endogenous activity control before attributing color to GBF1. |
| A nuclear or membrane-outline pattern dominates. | Those compartments conflict with HPA's granular cytoplasmic IHC profile and supported Golgi ICC-IF location (HPA tissue IHC; HPA subcellular ICC-IF). | Check counterstain and tissue morphology, then compare the suspect pattern with a no-primary control and an HPA-reported positive cell population (HPA tissue IHC; general IHC practice). Do not score compartment-mismatched color as a confirmed GBF1 positive. |
| Cells reported as not detected stain as strongly as the expected positives. | The discordance may reflect background or cross-reactivity, but HPA's tissue-IHC reliability is Uncertain (HPA tissue IHC). | Score each cell type separately and compare signal with a no-primary control and a documented medium population (HPA tissue IHC; general IHC practice). Treat disagreement as a reason to validate the catalog antibody in the specimen, not as proof that the HPA profile is universal. |
| Only a diffuse cytoplasmic haze remains after background checks. | GBF1 is listed in cytoplasm and cycles on and off Golgi membranes, although HPA describes granular cytoplasmic tissue staining (UniProt Q92538; HPA tissue IHC). | Assess whether reproducible granules or perinuclear enrichment can be resolved in the appropriate cells; compare the same compartments in controls (HPA tissue IHC; general IHC practice). If resolution is insufficient, record the result as cytoplasmic staining without claiming Golgi localization. |
| Can IF/ICC resolve an ambiguous Golgi-region IHC pattern? | HPA reports a Supported Golgi location by ICC-IF, whereas its tissue-IHC profile is Uncertain (HPA subcellular ICC-IF; HPA tissue IHC). | Use the supported ICC-IF localization as a compartment cross-check, not as proof that paraffin-section IHC is specific (HPA subcellular ICC-IF; HPA tissue IHC). Consult the separate IF/ICC guide for that application; evaluate this slide with IHC controls and cell-specific tissue evidence. |
Experiment evidence
IHC & IF images showing GBF1 staining
A01817-1 · IHCIHC analysis of GBF1 using anti-GBF1 antibody (A01817-1). GBF1 was detected in a paraffin-embedded section of human prostate cancer tissue.
A01817-1 · IHCIHC analysis of GBF1 using anti-GBF1 antibody (A01817-1). GBF1 was detected in a paraffin-embedded section of rat brain tissue. Heat mediate
A01817-1 · IFIF analysis of GBF1 using anti-GBF1 antibody (A01817-1). GBF1 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107862-GBF1/tissue/cerebral+cortex#imid_9406088
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000107862-GBF1/tissue/cerebellum#imid_9406083
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107862-GBF1/tissue/colon#imid_9408310
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107862-GBF1/tissue/liver#imid_9408327
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037759; green = GBF1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA037759; green = GBF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GBF1 IHC & IF
🧪Run adrenal gland first: glandular cells should stain (HPA: Medium in adrenal gland glandular cells). Use duodenum glandular cells as a negative comparator (HPA: Not detected); HPA identifies no negative cell population within the adrenal gland slide, so any unstained cells serve only as background comparators, not validated biological negatives.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GBF1 in A-431, U-251MG, U2OS, hTERT-RPE1 (serum starved), with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and rabbit IgG isotype control matched to the primary antibody’s clonality and concentration (selected-SKU caption: rabbit anti-GBF1); use GBF1 knockout material as a biological negative, or peptide competition if the immunizing peptide is available. Quench endogenous peroxidase and assess background before interpreting DAB staining (selected-SKU caption: peroxidase detection with DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected-SKU tissue-IHC caption does not state a fixative. The caption reports heat-mediated EDTA retrieval at pH 8.0 for a paraffin-section example, but does not establish a retrieval requirement for adrenal gland (selected-SKU tissue-IHC caption). IF is feasible in principle because Golgi localization is supported by ICC-IF images (HPA subcellular: Golgi apparatus), but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC or identify an adrenal-specific artefact.
HPA tissue IHC evidence for GBF1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | 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 → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GBF1 IHC Tips
Troubleshoot GBF1 staining in paraffin sections by checking retrieval, cytoplasmic localization, background controls, and scoring consistency (UniProt Q92538; HPA tissue IHC).
Which retrieval conditions should I start with for GBF1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for GBF1 paraffin sections (datasheet A01817-1). The selected tissue image used this retrieval before incubation with 2 μg/ml antibody overnight at 4°C, so keep those conditions together for an initial comparison (caption A01817-1). If staining is weak, vary heating and cooling time on adjacent sections while keeping the antibody concentration and detection method fixed (standard IHC practice). Check tissue preservation and an omission control before calling faint staining a retrieval failure (standard IHC practice). Record whether changes improve granular cytoplasmic staining without raising diffuse background (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or uneven GBF1 staining?
GBF1-specific fixation sensitivity is unknown from the supplied evidence; the selected image describes paraffin-embedded tissue but does not state its fixative (caption A01817-1). Compare sections with documented fixation histories under the same EDTA pH 8.0 retrieval and staining conditions (datasheet A01817-1; standard IHC practice). Inconsistent fixation or processing can alter antigen accessibility and tissue morphology, so inspect both before adjusting antibody concentration (standard IHC practice). Use an internal staining reference and an omission control on each run to separate processing variation from detection background (standard IHC practice). Do not assign a GBF1-specific fixation effect without a controlled comparison (standard IHC practice).
What staining pattern is plausible, and when should I suspect mislocalization?
Expect granular cytoplasmic staining in tissue sections, with possible concentration near the Golgi, rather than a uniformly nuclear pattern (HPA tissue IHC; HPA subcellular). GBF1 cycles between cytoplasm and early Golgi membranes and is also annotated at the ER–Golgi intermediate compartment and trans-Golgi network (UniProt Q92538). Its rapid membrane recruitment and release mean that a single chromogenic section cannot resolve every trafficking state (UniProt Q92538; standard IHC interpretation). Compare similarly processed cells within the same section before treating differences in granule prominence as biological changes (standard IHC practice). Strong nuclear-only or luminal staining warrants review of morphology, background controls, and antibody specificity (UniProt Q92538; standard IHC practice).
How do isoforms and epitope location affect interpretation of a negative section?
GBF1 has 4 annotated isoforms and a SEC7 domain at residues 693–883 (UniProt Q92538). The supplied antibody caption does not identify its epitope, so a negative section cannot establish that all isoforms are absent (caption A01817-1; UniProt Q92538). Check the antibody's documented immunogen or epitope against isoform sequences before making an isoform-specific claim (standard IHC practice). GBF1 has no transmembrane segment and has multiple annotated phosphorylation sites, but neither fact establishes how this antibody recognizes processed tissue (UniProt Q92538). Compare retrieval and staining controls before interpreting a weak result as epitope loss (standard IHC practice).
How should I assess GBF1 in multiplex IF alongside the IHC findings?
For a separate IF experiment, pair GBF1 with a marker identifying the expected cell population, such as an epithelial marker when examining glandular cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with lower tissue autofluorescence, verified using an unstained control (standard IF practice). GBF1 lacks a transmembrane segment and associates with Golgi membranes, but the catalog antibody's epitope side is unspecified (UniProt Q92538; caption A01817-1). Test mild permeabilisation for access to a cytoplasmic epitope and assess whether stronger treatment disrupts granular localization (standard IF practice). Validate IF staining independently; the selected antibody evidence describes chromogenic IHC in paraffin sections (caption A01817-1).
How can I reduce diffuse or granular background without losing GBF1 signal?
The selected IHC image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary before DAB development (caption A01817-1). Begin with a primary-omission control and a peroxidase block to assess nonspecific detection and endogenous enzyme activity (standard IHC practice). If diffuse color persists, titrate the primary antibody and review wash stringency on matched sections while keeping development time constant (standard IHC practice). Distinguish isolated DAB precipitate from the granular cytoplasmic pattern reported for GBF1 (HPA tissue IHC; standard IHC practice). Check section edges and damaged areas before attributing unusually intense deposits to GBF1 (standard IHC practice).
What should I measure when GBF1 staining varies across a section? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and region of interest before scoring granular cytoplasmic GBF1 staining (HPA tissue IHC; standard IHC practice). For comparable cell populations, report percentage of positive cells plus an H-score from 0–300 based on staining intensity and the fraction at each intensity (standard IHC practice). If the question concerns stained-cell abundance, report positive-cell density per mm² of viable tissue instead (standard IHC practice). Normalize to the number of eligible cells or viable tissue area, and hold retrieval, DAB development, and image thresholds constant across sections (standard IHC practice). Record excluded necrotic and edge regions before comparing groups (standard IHC practice).
Which findings support true GBF1 staining rather than an artefact?
A credible result combines granular cytoplasmic staining with appropriate tissue morphology and a clean primary-omission control (HPA tissue IHC; standard IHC practice). Golgi-associated signal is consistent with GBF1 biology, whereas strong nuclear-only staining needs additional specificity checks (HPA subcellular; UniProt Q92538). Judge the stained cell population explicitly: HPA reports medium staining in several glandular cell populations but rates its tissue IHC reliability uncertain (HPA tissue IHC). Exclude necrotic areas, section edges, and deposits reproduced in the omission control; assess endogenous peroxidase when DAB signal persists without primary antibody (standard IHC practice). Treat isolated staining differences cautiously until they reproduce across sections and runs (standard IHC practice).
Boster reagents
Best GBF1 / Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 IHC Antibodies
The IHC-validated antibody has paraffin-section images from human prostate and breast cancer tissue and rat and mouse brain (catalog IHC captions); IF data show U2OS cells (catalog IF caption).
Real IHC data
Anti-GBF1 Antibody ®
Cat # A01817-1
A01817-1 is listed for IHC, ICC and IF in human, mouse and rat samples (catalog applications and reactivity). Its IHC captions show human prostate and breast cancer tissue and rat and mouse brain paraffin sections; its IF caption shows U2OS cells (catalog image captions).
Which to pick: Choose A01817-1 for paraffin-section IHC: its own caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody in human prostate cancer tissue (A01817-1 IHC caption); the fixative is unreported (A01817-1 IHC caption). For IF/ICC, the same SKU has a U2OS cell image at 5 μg/ml (A01817-1 IF caption). For cross-species IHC, it lists human, mouse and rat reactivity and has paraffin-section images for all three; the host is rabbit and clonality is unspecified (catalog reactivity, host and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92538 (GBF1_HUMAN, Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1).
- Human Protein Atlas. GBF1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. GBF1 subcellular location (ICC-IF): Localized to the Golgi apparatus..
- Human Protein Atlas. GBF1 antibody validation summary (2 antibodies).
- Intracellular protein GBF1 displays significant associations with amyloid pathology in Alzheimer's disease. Alzheimer's & dementia : the journal of the Alzheimer's Association 2026 — PMC13084705.
- PubMed PMID:9828135 — UniProt-cited evidence.
- PubMed PMID:9039502 — UniProt-cited evidence.
- PubMed PMID:12168954 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GBF1 (Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1). Expected localisation Granular cytoplasm in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01817-1). Validated IHC antibody: A01817-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A01817-1); 2. Fixation: Image fixative and duration unreported (datasheet A01817-1); 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 A01817-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01817-1); 8. Primary antibody: Rabbit anti-GBF1, 2-5 μg/ml (datasheet A01817-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01817-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01817-1); Caveat: Tissue staining has only medium RNA concordance (HPA tissue IHC); Isoform / epitope: 4 isoforms; epitope coverage is unspecified (UniProt); Regulation: Ubiquitous expression (UniProt). Evidence: UniProt Q92538, HPA tissue IHC · Uncertain, HPA ICC-IF · Golgi apparatus., Datasheet A01817-1, PMC13084705. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GBF1/immunohistochemistry-ihc-guide.