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GBP4 / Guanylate-binding protein 4 · IHC design guide
Design Immunohistochemistry for GBP4
Plan chromogenic IHC for GBP4 in paraffin sections using the catalog antibody at 1:100–1:300 (datasheet: A11011). Assess cytoplasmic staining in glandular or kidney tubular cells, with adipocytes as a low-signal comparison (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GBP4 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 in several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in glandular and kidney tubular cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | GBP1/2/5 heterodimers can shift GBP4 location (UniProt) | |
| Regulation | IFN-inducible (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–640 chain (UniProt) |
Section 1
Recommended GBP4 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 2 published GBP4 IHC methods for ovarian and pancreatic cancer tissue (PMC8155613; PMC11306721).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A11011) |
| Fixation | Image fixative and duration unreported (datasheet A11011); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit monoclonal (clone 3A8) anti-GBP4, 1:100-1:300 (datasheet A11011) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GBP4-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page antigen retrieval); the published excerpts provide no retrieval conditions (PMC8155613; PMC11306721).
Section 2
What Is the Expected GBP4 Staining Pattern?
GBP4 is reported in the cytoplasm, perinuclear region, Golgi membrane and nucleus, with no transmembrane segment (UniProt Q96PP9 localization and topology). In paraffin sections, expect cytoplasmic staining in HPA-positive glandular cells, kidney tubules and lymph node non-germinal center cells (HPA tissue IHC). HPA rates its tissue IHC evidence Enhanced, with medium consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Cytoplasmic staining in colon or duodenal glandular cells, kidney tubules, or lymph node non-germinal center cells (HPA tissue IHC). | This matches the reported high-staining cell populations (HPA tissue IHC). Assess the named cells against nearby cells and the counterstain; a perinuclear accent is compatible with GBP4 localization but is not required to call a paraffin section positive (UniProt Q96PP9 localization). |
| Dominant staining in an unexpected compartment, with little staining in the expected cytoplasm. | Check section morphology and detection artefact before assigning that pattern to GBP4. HPA reports cytoplasmic tissue staining, while UniProt also lists nuclear and Golgi-associated localization; nuclear signal alone cannot be ruled out categorically from these sources (HPA tissue IHC; UniProt Q96PP9 localization). |
| Strong signal in adipocytes, hippocampal glial cells, or ovarian stromal cells. | These named populations were not detected in HPA tissue IHC (HPA tissue IHC). Review antibody specificity and endogenous chromogenic activity, especially if the positive-control cell pattern is absent. A signal in another, unlisted cell population is not automatically cross-reactivity: HPA reports low tissue RNA specificity (HPA tissue IHC). |
| Diffuse chromogen across cells and extracellular space, without a readable cellular pattern. | Treat the distribution as background until controls establish otherwise. Review blocking, washes, primary-antibody concentration and detection controls as general IHC practice; HPA's cytoplasmic tissue profile does not validate uniform extracellular staining (HPA tissue IHC). |
| No staining in the expected cells of a known-positive section. | Colon glandular cells and kidney tubule cells are reported as high-staining examples (HPA tissue IHC). First check that those cells are present and interpretable; then review the antibody's IHC-P instructions and detection controls. A blank section alone cannot establish GBP4 absence. |
💡Expected GBP4 appearanceCall a positive result when the expected glandular, tubular or non-germinal center cells show interpretable cytoplasmic chromogen, strongest in HPA high-staining examples; diffuse extracellular colour or dominant signal in HPA not-detected cells warrants a control review (HPA tissue IHC).
| Cell population and comparison tissue | HPA reports high staining in several glandular populations, kidney tubules and lymph node non-germinal center cells, but no detection in three specified cell populations (HPA tissue IHC). Score the named cells; a whole-tissue positive or negative label can hide that distinction. |
| Intracellular distribution | Golgi, cytoplasmic, perinuclear and nuclear locations are listed by UniProt; GBP1, GBP2 and GBP5 heterodimers can alter GBP4 location (UniProt Q96PP9 localization and subunit). HPA tissue IHC describes cytoplasmic expression, so do not require a sharply defined Golgi pattern in chromogenic sections (HPA tissue IHC). |
| Strength of the evidence | The tissue IHC profile is Enhanced, with medium staining-to-RNA consistency (HPA tissue IHC reliability). Antibody HPA030101 has Enhanced IHC validation, whereas HPA062039 has Supported ICC validation (HPA antibody validation); these are application-specific assessments. |
| Chromogenic detection controls | Endogenous enzyme activity can create colour unrelated to primary-antibody binding (general IHC practice). Check a suitable detection control when staining is widespread or appears in HPA not-detected cells; that observation does not, by itself, identify the cause (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected cells stain weakly in a positive-control paraffin section. | The reported HPA level is an observed tissue pattern, not a promised intensity under every staining setup (HPA tissue IHC). The run may lack adequate detectable contrast. | Verify cell identity and counterstain, then review the catalog antibody's IHC-P instructions for retrieval, dilution and detection settings. Adjust one general IHC variable at a time against the same positive control; no GBP4-specific retrieval setting is supplied here. |
| Colour appears chiefly outside cells or uniformly across the section. | A widespread pattern does not match HPA's cytoplasmic tissue profile (HPA tissue IHC); nonspecific detection or residual chromogen can obscure cellular boundaries (general IHC practice). | Inspect the no-primary detection control, blocking and washes. Reassess staining only where intact cells and their counterstained boundaries can be identified (general IHC practice). |
| Adipocytes, hippocampal glial cells or ovarian stromal cells appear strongly positive. | HPA records these specific populations as not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible, but the staining alone cannot distinguish them. | Compare a positive tissue and a no-primary control in the same run. If unexpected staining persists only with primary antibody, examine IHC validation and repeat at an appropriately optimized antibody dilution (general IHC practice). |
| The section looks predominantly nuclear, without convincing cytoplasmic staining. | UniProt includes nuclear localization, but HPA describes cytoplasmic staining in several tissues (UniProt Q96PP9 localization; HPA tissue IHC). A nuclear-only slide therefore needs context, not an automatic positive call. | Check morphology, counterstain and detection controls, then compare the same cell type in an HPA high-staining tissue. Record nuclear staining separately from the expected cytoplasmic pattern (HPA tissue IHC). |
| A tissue seems negative despite visible staining elsewhere on the slide. | HPA levels refer to particular cell populations; low-staining cells include hepatocytes and splenic red-pulp cells (HPA tissue IHC). Section-wide scoring can misclassify a sparse cell-specific pattern. | Identify and score the relevant cells individually, then compare with a named high-staining control population. Do not infer failure solely from weak signal in an HPA low-staining cell type (HPA tissue IHC). |
| Can the paraffin IHC pattern be used as the IF/ICC protocol or its expected image? | HPA ICC-IF places GBP4 mainly at the Golgi (supported) and additionally at the plasma membrane (uncertain); its image cell lines are HEL, RT-4 and U2OS (HPA subcellular ICC-IF). Those observations use a different application from tissue IHC. | Use the separate IF/ICC guide for that application. Interpret a Golgi-enriched IF signal with the ICC evidence, and treat plasma-membrane signal cautiously; do not turn the tissue IHC pattern into an IF/ICC protocol (HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing GBP4 staining
A11011 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma, using GBP4 Antibody. The picture on the right is blocked with the
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162654-GBP4/tissue/cerebral+cortex#imid_7628880
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000162654-GBP4/tissue/duodenum#imid_7628805
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162654-GBP4/tissue/colon#imid_7628814
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162654-GBP4/tissue/liver#imid_7628823
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA062039; green = GBP4, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA062039; green = GBP4, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GBP4 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, assess adjacent nonglandular cells for background staining without assuming they are GBP4-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show GBP4 in HEL, RT-4, U2OS, with annotated localisation: Golgi apparatus (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls matched to the primary antibody’s host species and clonality (standard IHC practice). Use a peptide-block control for the catalog antibody (A11011 tissue-IHC caption: staining was peptide-blocked), and quench endogenous peroxidase in the colon section before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the A11011 paraffin-section caption does not state a fixative (A11011 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier than paraffin IHC; HPA reports ICC-IF images in HEL, RT-4 and U2OS, but no tissue IF result is supplied (HPA subcellular). In colon, inflammatory cells can contribute endogenous peroxidase background, so interpret glandular staining against the controls (standard IHC practice).
HPA tissue IHC evidence for GBP4
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced GBP4 IHC Tips
Use compartment, cell type, and matched controls to troubleshoot GBP4 staining in paraffin sections; assess IF separately.
How should I troubleshoot weak GBP4 staining after antigen retrieval?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Keep section thickness, cooling, and detection conditions consistent while comparing runs, so a change in signal can be attributed to retrieval (standard IHC practice). Include a known positive section, such as kidney tubules with high reported expression (HPA: High in kidney tubular cells). If staining remains weak, compare a gentler retrieval condition on adjacent sections and check whether tissue morphology and background improve (standard IHC practice). The A11011 breast carcinoma caption documents paraffin sections but does not report a fixative or retrieval method (A11011 caption).
Could fixation explain weak or uneven GBP4 staining?
GBP4-specific sensitivity to fixation is unknown from the supplied evidence; the A11011 image caption identifies paraffin-embedded breast carcinoma but does not state its fixative (A11011 caption). Record the actual fixative, fixation duration, processing history, and section age for each specimen before comparing staining (standard IHC practice). Run sections with different processing histories in the same staining batch, alongside a reference section, to distinguish specimen effects from reagent drift (standard IHC practice). Excessively harsh processing can reduce accessible epitopes in general, so assess morphology and control staining before changing retrieval (standard IHC practice). Do not infer fixation tolerance from GBP4 localisation or processing annotations (UniProt Q96PP9).
Which GBP4 staining patterns should I expect in paraffin sections?
Expect predominantly cytoplasmic tissue staining as a starting pattern (HPA: cytoplasmic expression in several tissues). GBP4 is also annotated in the Golgi apparatus, nucleus, and perinuclear cytoplasm, with localisation influenced by heterodimer partners (UniProt Q96PP9 subcellular location; PubMed:21151871). Assess a discrete perinuclear or Golgi-like signal against overall cytoplasmic staining, tissue morphology, and a no-primary control before assigning it to GBP4 (standard IHC practice). HPA supports Golgi localisation in cell imaging, while its additional plasma-membrane localisation is uncertain (HPA subcellular). Diffuse staining across every tissue compartment, especially where morphology is damaged, warrants a background check before biological interpretation (standard IHC practice).
How can epitope accessibility affect GBP4 staining?
Confirm the immunogen or mapped epitope for the exact antibody before attributing a weak result to GBP4 biology (standard IHC practice). The supplied GBP4 record lists a single 1–640 chain, no annotated isoforms, no transmembrane segment, and no glycosylation sites (UniProt Q96PP9 processing, isoforms, topology, glycosylation). These annotations do not establish whether this antibody’s epitope survives tissue processing (UniProt Q96PP9; A11011 caption: fixative unreported). Compare adjacent sections under the stated retrieval condition and evaluate staining with appropriate positive and no-primary controls (page retrieval setting; standard IHC practice). A peptide-blocked image accompanies the A11011 paraffin breast carcinoma example; interpret it as evidence for that image’s antibody-reactive signal, not universal tissue specificity (A11011 caption).
How should I adapt GBP4 localisation checks to multiplex IF?
For the separate IF/ICC workflow, pair GBP4 with a marker identifying the cell population being evaluated, then inspect each channel and the merged image (standard IF practice). Choose fluorophores after measuring tissue autofluorescence and favor a spectrally distinct, brighter channel for the weaker target signal (standard IF practice). GBP4 has no transmembrane segment and is annotated in cytoplasm and nucleus, so validate permeabilisation for access to intracellular epitopes (UniProt Q96PP9 topology and subcellular location; standard IF practice). Include single-stain and no-primary controls to assess bleed-through and nonspecific fluorescence (standard IF practice). HPA cell images support a principal Golgi localisation; its additional plasma-membrane assignment is uncertain (HPA subcellular).
What should I check when GBP4 chromogenic staining is widespread?
Compare the stained section with no-primary and matched detection controls before calling diffuse chromogen a GBP4 signal (standard IHC practice). Check endogenous peroxidase blocking, secondary-antibody background, chromogen development, and incomplete washing as possible causes of widespread colour (standard chromogenic IHC practice). Examine tissue edges, folds, damaged areas, and necrosis separately because these regions can retain reagents or show misleading deposits (standard IHC practice). High GBP4 expression is reported in kidney tubular cells and several glandular cell populations, whereas adipocytes are reported as not detected (HPA tissue IHC). Use those cell-level patterns as contextual checks, while allowing for specimen-specific biology and the HPA profile’s medium RNA–staining consistency (HPA tissue IHC).
How should I score GBP4 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cell population before scoring, and apply the same rule to every section (standard IHC practice). For cellular staining, report the percentage of positive eligible cells and an H-score using intensity grades 0–3, giving a 0–300 range (standard IHC scoring practice). If the question concerns infiltrating positive cells, report their density per mm² of viable tissue instead (standard IHC quantification practice). Normalize counts to the corresponding eligible cells or viable tissue area, and exclude folds, edges, and necrosis using a predefined rule (standard IHC practice). Score cytoplasmic and distinct perinuclear patterns separately when resolution permits (HPA tissue IHC; UniProt Q96PP9 subcellular location).
How can I distinguish true GBP4 positivity from artefact?
Look for staining in intact cells with a plausible cytoplasmic or perinuclear distribution, then compare cell identity and location with matched controls (HPA tissue IHC; UniProt Q96PP9 subcellular location; standard IHC practice). Kidney tubular cells and several glandular populations are reported high, while adipocytes, hippocampal glia, and ovarian stromal cells are reported not detected (HPA tissue IHC). Treat staining confined to section edges, necrotic regions, or areas positive in the no-primary control as suspect (standard IHC practice). Check endogenous enzyme activity when chromogen appears independently of the primary antibody (standard chromogenic IHC practice). The A11011 peptide-blocked paraffin-section image supports interpretation of that example, but specimen-level conclusions still require appropriate controls (A11011 caption; standard IHC practice).
Boster reagents
Best GBP4 / Guanylate-binding protein 4 IHC Antibodies
Anti-GBP4 A11011 has an IHC image from paraffin-embedded human breast carcinoma (catalog image caption); IF is listed, but no IF image is provided (catalog applications; catalog IF image alts).
Real IHC data
Anti-Guanylate-binding protein 4 GBP4 Antibody
Cat # A11011
A11011 will render with an IHC image of paraffin-embedded human breast carcinoma, including a peptide-blocked comparison (catalog image caption). IHC and IF are listed applications, and Human and Mouse are listed reactivities; no IF image is provided (catalog applications; catalog reactivity; catalog IF image alts).
Which to pick: For tissue IHC, choose A11011: its own image shows paraffin-embedded human breast carcinoma (catalog image caption), and its IHC dilution is 1:100–1:300 (datasheet: IHC dilution). The fixative is unreported (catalog image caption). For IF/ICC or human–mouse work, A11011 is the only listed option: it is rabbit clone 3A8, lists IF as an application and Human and Mouse as reactivities, but provides no IF image or ICC validation (catalog host and clone; catalog applications; catalog reactivity; catalog IF image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96PP9 (GBP4_HUMAN, Guanylate-binding protein 4).
- Human Protein Atlas. GBP4 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. GBP4 subcellular location (ICC-IF): Mainly localized to the Golgi apparatus. In addition localized to the plasma membrane..
- Human Protein Atlas. GBP4 antibody validation summary (2 antibodies).
- NLRP3 activation by lipopolysaccharide (LPS) mediates the pyroptosis of human spermatogonial stem cells via GBP4 regulation. Asian journal of andrology 2026 — PMC13423383.
- Identification of a Prognostic Signature for Ovarian Cancer Based on the Microenvironment Genes. Frontiers in genetics 2021 — PMC8155613.
- DNA hypo-methylation and expression of GBP4 induces T cell exhaustion in pancreatic cancer. Cancer immunology, immunotherapy : CII 2024 — PMC11306721.
- Integrated transcriptome and single-cell RNA sequencing analysis revealed the prognostic significance of GBP4 in pancreatic adenocarcinoma. Translational oncology 2025 — PMC12496431.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:12693554 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GBP4 (Guanylate-binding protein 4). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A11011 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A11011); 2. Fixation: Image fixative and duration unreported (datasheet A11011); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit monoclonal (clone 3A8) anti-GBP4, 1:100-1:300 (datasheet A11011); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: GBP1/2/5 heterodimers can shift GBP4 location (UniProt); Isoform / epitope: No annotated isoforms; one 1–640 chain (UniProt); Regulation: IFN-inducible (UniProt). Evidence: UniProt Q96PP9, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the Golgi apparatus. In addition localized to…, Datasheet A11011, PMC8155613, PMC11306721. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GBP4/immunohistochemistry-ihc-guide.