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GBP5 / Guanylate-binding protein 5 · IHC design guide
Design Immunohistochemistry for GBP5
Plan GBP5 staining in paraffin sections around cytoplasmic signal in a subset of immune cells (HPA tissue IHC). Use lung macrophages as a positive tissue reference and account for IFN-inducible expression when interpreting intensity (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GBP5 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 | Cytoplasm in a subset of immune cells (HPA tissue IHC) | |
| Staining pattern | Scattered immune cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Gallbladder+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 sections (standard IHC practice; not target-specific) | |
| Caveat | Sparse positive cells may be missed in sampled fields (HPA tissue IHC) | |
| Regulation | IFN-inducible (UniProt) | |
| Isoform / epitope | Two isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended GBP5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published GBP5 IHC methods for placental tissue, intestine, and NSCLC tissue microarrays (PMC13093737; PMC9201962; PMC10183185).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07110); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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 anti-GBP5, 1:100–1:500 starting range (standard) |
| 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 | GBP5-positive staining in glandular cells of gallbladder (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a subset of immune cells in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval setting); compare EDTA retrieval when adapting the placental or NSCLC methods (PMC13093737; PMC10183185).
Section 2
What Is the Expected GBP5 Staining Pattern?
GBP5 should appear mainly cytoplasmic, sometimes punctate or concentrated near intracellular membranes, consistent with its cytoplasmic, vesicle membrane and Golgi membrane locations and lack of a transmembrane segment (UniProt Q96PP8 topology and subcellular location). In tissue sections, expect staining in a subset of immune cells, with some other HPA-listed cell types also positive (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between staining and RNA data (HPA tissue IHC).
| Cytoplasmic stain in scattered lung macrophages or cells in splenic red pulp, around the reported Medium level (HPA tissue IHC). | This fits the reported cell distribution and GBP5's cytoplasmic and intracellular membrane locations (HPA tissue IHC; UniProt Q96PP8 subcellular location). Assess the named cells rather than expecting every cell in the section to stain (HPA tissue IHC). |
| Predominantly nuclear staining, with little cytoplasmic signal. | An isolated nuclear pattern does not match the listed GBP5 locations (UniProt Q96PP8 subcellular location). Treat it as suspect and check controls and detection conditions before interpreting it as target staining (standard IHC practice). |
| Strong signal in adipocytes or adrenal gland glandular cells. | These named cell populations were Not detected by HPA tissue IHC; staining there merits a cross-reactivity or endogenous detection-activity check (HPA tissue IHC; standard IHC practice). Cell identity matters: gallbladder glandular cells were Medium, so glandular staining alone is not disqualifying (HPA tissue IHC). |
| Diffuse stain across tissue and empty areas, without a discernible cellular pattern. | That distribution cannot establish the reported cytoplasmic, cell-selective pattern (HPA tissue IHC). Review background and no-primary controls for nonspecific binding or detection background before scoring cells (standard IHC practice). |
| No signal in lung macrophages on a section expected to contain them. | HPA reports Medium staining in lung macrophages, so absent signal calls for a run check; it does not alone prove GBP5 is absent from the specimen (HPA tissue IHC). Confirm the cells are present, then review the IHC workflow and controls (standard IHC practice). |
💡Expected GBP5 appearanceCall a result positive when identifiable HPA-listed cells show cytoplasmic staining, sometimes with intracellular puncta, at an intensity compatible with the reported Medium examples; isolated nuclear staining or indiscriminate background is suspect (HPA tissue IHC; UniProt Q96PP8 subcellular location; standard IHC practice).
| Cell selection for a positive comparison | Lung macrophages, splenic red-pulp cells and tonsillar non-germinal-center cells are reported Medium; choose a section containing the specified cells and score them directly (HPA tissue IHC). |
| Interpretation of epithelial staining | HPA reports Medium staining in gallbladder glandular cells and urinary-bladder urothelial cells, alongside immune-cell staining; epithelial signal needs tissue and cell context (HPA tissue IHC). |
| Location and topology | GBP5 is listed in cytoplasm and on cytoplasmic vesicle and Golgi membranes, with no transmembrane segment; intracellular granular or perinuclear accentuation can fit this distribution (UniProt Q96PP8 subcellular location and topology). |
| Strength of tissue evidence | HPA calls the tissue pattern Supported and describes medium consistency with RNA expression; this supports comparison with its images but leaves room for specimen-to-specimen differences (HPA tissue IHC). |
| Antibody evidence | The listed rabbit polyclonal antibody HPA028656 has Supported IHC status; the payload does not report Enhanced IHC validation or provide a catalog-antibody dilution (HPA antibodies). |
| Isoforms and processing | Two isoforms and a chain spanning residues 1–583 are listed, but no antibody epitope is supplied; the record cannot establish whether this stain distinguishes isoforms or processing states (UniProt Q96PP8 isoforms and processing). |
| IF/ICC Q&A: Is a GBP5 IF pattern established here? | No HPA ICC-IF image or main subcellular location is available; the UniProt locations are a biological expectation, not an image-validated IF pattern or IF protocol (HPA subcellular ICC-IF; UniProt Q96PP8 subcellular location). |
| Situation | Likely cause | Next action |
|---|---|---|
| A positive comparison section appears blank. | The selected section may lack the relevant HPA-listed cells, or the IHC run may have failed; the image alone cannot distinguish these possibilities (HPA tissue IHC; standard IHC practice). | Verify cell presence on the counterstain and compare a separate known-positive section processed in the same run; then review detection controls (HPA tissue IHC; standard IHC practice). |
| Only nuclei are colored. | A nuclear-only pattern conflicts with the recorded cytoplasmic and intracellular membrane locations and may represent nonspecific staining (UniProt Q96PP8 subcellular location; standard IHC practice). | Inspect a no-primary control and compare the staining compartment in HPA-listed positive cells before accepting the signal (standard IHC practice; HPA tissue IHC). |
| Adipocytes stain broadly. | HPA lists adipocytes in adipose tissue as Not detected; broad staining there may reflect nonspecific antibody binding or endogenous detection activity (HPA tissue IHC; standard IHC practice). | Check no-primary and detection controls, then compare staining in an HPA-listed positive cell population under the same run conditions (standard IHC practice; HPA tissue IHC). |
| Background obscures individual cells. | Diffuse background prevents assessment of the cell-selective cytoplasmic pattern HPA describes; its precise source cannot be inferred from this record (HPA tissue IHC; standard IHC practice). | Use no-primary and detection controls to localize background, then review blocking, washes and detection conditions as general IHC workflow checks (standard IHC practice). |
| Only a few cells stain in an otherwise pale section. | Sparse positive cells can fit HPA's description of cytoplasmic expression in a subset of immune cells; tissue-wide uniformity is not the reported pattern (HPA tissue IHC). | Identify the stained cells on the counterstain and compare the same cell type with HPA's tissue images before calling the run weak (HPA tissue IHC; standard IHC practice). |
| IF shows puncta but lacks a matching HPA ICC-IF image. | UniProt lists vesicle and Golgi membrane locations, while HPA supplies no ICC-IF images or main-location call for GBP5 (UniProt Q96PP8 subcellular location; HPA subcellular ICC-IF). | Describe the IF observation as provisional and assess appropriate IF controls; use this page's HPA evidence for IHC tissue interpretation, not IF image validation (HPA tissue IHC; HPA subcellular ICC-IF; standard IF practice). |
Experiment evidence
IHC & IF images showing GBP5 staining
A07110 · IHCImmunohistochemistry of GBP5 in human liver tissue with GBP5 antibody at 2.5 μg/ml.
A07110 · IFImmunofluorescence of GBP5 in human liver tissue with GBP5 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000154451-GBP5/tissue/cerebral+cortex#imid_20604819
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000154451-GBP5/tissue/colon#imid_20604737
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000154451-GBP5/tissue/liver#imid_20604754
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000154451-GBP5/tissue/kidney#imid_20604747
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GBP5 IHC & IF
🧪Run lung first and look for staining in macrophages (HPA: lung macrophages, Medium). Run adipose tissue as the negative comparator and score adipocytes for absent staining (HPA: adipocytes, Not detected); on the lung slide, use neighboring cells without specific chromogenic deposit as internal background comparators (standard IHC practice).
Positive control tissue: Gallbladder (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GBP5; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched isotype control of the primary antibody’s host species and clonality; confirm specificity with a GBP5 knockout specimen or a peptide block if a suitable immunizing peptide is available (standard IHC practice). Lung macrophage pigment can resemble chromogenic signal, so compare its appearance with the control slides (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A07110 tissue-IHC caption also does not report its fixative (selected-SKU caption). Use the catalog antibody’s IHC-P protocol as the starting point for paraffin-section retrieval (application evidence: IHC-P protocol). Frozen-section or IF staining is not established as easier here, and lung macrophage pigment may complicate chromogenic scoring (HPA: no ICC-IF images; standard IHC practice).
HPA tissue IHC evidence for GBP5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Medium | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Urinary bladder | Urothelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GBP5 IHC Tips
Troubleshoot GBP5 staining in paraffin section chromogenic IHC by checking retrieval, cell identity, intracellular pattern, and assay controls before interpreting signal.
How should I retrieve GBP5 in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat retrieval at 95–98 °C for 20 minutes (page retrieval rule). Cool sections consistently, then compare GBP5 signal and tissue preservation against a section processed without retrieval. If staining remains weak, test an EDTA buffer at pH 9.0 as a fallback on matched sections, holding antibody concentration and detection time constant. Inspect both expected cytoplasmic signal in immune cell subsets (HPA: tissue IHC profile) and nonspecific staining before adopting the alternative. The selected antibody image reports human liver staining at 2.5 µg/ml, but its caption does not specify retrieval conditions (catalog antibody A07110 caption).
Could fixation explain inconsistent GBP5 staining between paraffin blocks?
GBP5 specific sensitivity to fixation is unknown from the supplied evidence; the selected human liver IHC caption does not state a fixative (catalog antibody A07110 caption). For a controlled comparison, cut adjacent sections and keep deparaffinisation, citrate pH 6.0 retrieval, and chromogenic development consistent across blocks (page retrieval rule). Record each block’s fixation and processing history, then compare signal in the same cell population rather than across different tissue compartments. Include a known staining section in each run to distinguish run variation from block variation. Do not infer fixation sensitivity from tissue staining patterns (HPA: tissue IHC profile) or GBP5 topology (UniProt Q96PP8 topology).
Where should GBP5 staining appear, and what pattern warrants investigation?
Assess staining primarily in the cytoplasm, allowing a vesicular or perinuclear membrane associated pattern because GBP5 is annotated in cytoplasm, cytoplasmic vesicle membrane, and Golgi membrane (UniProt Q96PP8 subcellular location). GBP5 has no transmembrane segment, so membrane association alone does not identify an antibody’s epitope side (UniProt Q96PP8 topology). In tissue, compare staining with the reported cytoplasmic signal in subsets of immune cells (HPA: tissue IHC profile). Isolated nuclear staining should prompt review of counterstain, detection background, and the no primary control. Compare serial sections under identical development conditions before assigning a faint punctate pattern to a specific organelle.
How can isoforms and terminal processing affect interpretation of GBP5 IHC?
GBP5 has 2 annotated isoforms, but the supplied antibody information does not map its epitope to either one (UniProt Q96PP8 isoforms; catalog antibody A07110 caption). The annotated chain spans residues 1–583, with a 584–586 propeptide and a modified cysteine at residue 583 (UniProt Q96PP8 processing; modified residues). Check the antibody’s immunogen or epitope documentation before claiming isoform specificity or attributing weak staining to terminal processing. If that information remains unavailable, report the result as antibody detected GBP5 immunoreactivity, without assigning an isoform. Confirm a disputed pattern with an independently validated reagent or a biologically appropriate loss of signal control.
How should I adapt GBP5 localisation checks to multiplex immunofluorescence?
For the separate IF/ICC workflow, pair GBP5 with a macrophage marker when examining the reported macrophage positive lung compartment (HPA: Medium in lung macrophages). Choose a far red GBP5 fluorophore and measure an unstained section’s autofluorescence before setting exposure or spectral separation. GBP5 is cytoplasmic and associates with vesicle and Golgi membranes, but its antibody epitope side is unspecified (UniProt Q96PP8 subcellular location; catalog antibody A07110 caption). Optimise mild permeabilisation on matched samples so a cytoplasm facing epitope is accessible, without assuming access to a lumen facing epitope. Include single colour and no primary controls to distinguish overlap from bleed through and tissue background.
How do I separate true GBP5 signal from chromogenic background?
In paraffin section IHC, compare the antibody stained section with a no primary control after the same peroxidase block and DAB development; those steps are general chromogenic workflow controls. Review signal in cell populations reported as not detected, such as adipocytes in adipose tissue, alongside a suitable positive population (HPA: adipose adipocytes Not detected; lung macrophages Medium). Reduce excessive DAB development or antibody concentration only after checking whether the positive compartment remains visible. The selected image uses 2.5 µg/ml in human liver, without reporting its detection chemistry (catalog antibody A07110 caption). Diffuse staining of every cell, tissue edges, or pigment deposits merits control review before biological interpretation.
What GBP5 IHC scoring method best handles mixed immune cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, since GBP5 staining is reported in subsets of immune cells rather than uniformly across tissue (HPA: tissue IHC profile). For a cell based readout, record the percentage of positive cells and an H-score from 0–300, using fixed intensity categories and identical DAB development. For infiltrates, count GBP5 positive cells per mm² of viable tissue and also report their fraction of the annotated immune cell population. Normalise comparisons to that population or measured viable area, rather than whole section area when cellular composition differs. Exclude folds, necrosis, and edge staining using rules set before reviewing groups.
When is a GBP5 positive IHC result biologically credible?
A credible result places signal in the cytoplasm of an appropriate cell population, consistent with GBP5 localisation and the reported immune cell subset pattern (UniProt Q96PP8 subcellular location; HPA: tissue IHC profile). Lung macrophages offer a reported medium staining comparator, while adipose adipocytes are reported as not detected (HPA: lung macrophages Medium; adipose adipocytes Not detected). Check whether staining persists away from section edges and necrosis, and whether the no primary control excludes endogenous enzyme or pigment related signal. Predominantly nuclear staining or uniform staining across unrelated cells needs additional validation before calling it GBP5. Interpret borderline findings cautiously because the tissue IHC evidence has supported reliability with medium staining and RNA consistency (HPA: reliability description).
Boster reagents
Best GBP5 / Guanylate-binding protein 5 IHC Antibodies
GBP5 antibody A07110 has pictured IHC and IF staining in human liver tissue (catalog: A07110 image captions). Its listed reactivity covers human, mouse, and rat (catalog: A07110 reactivity).
Real IHC data
Anti-Guanylate-binding protein 5 GBP5 Antibody
Cat # A07110
A07110 is the sole rendered card; its captions show IHC in human liver tissue at 2.5 μg/ml and IF in human liver tissue at 20 μg/ml (catalog: A07110 image captions). IHC-P and IF are listed applications, but the supplied staining images document human tissue only (catalog: A07110 applications and image captions).
Which to pick: For tissue IHC, pick A07110: IHC-P is listed, and its own caption shows human liver staining at 2.5 μg/ml; the fixative is unreported (catalog: A07110 applications and IHC image caption). For IF, A07110 has a human liver image; for ICC, A07110-1 lists ICC and IF at 1:50–400 but has no supplied staining image (catalog: A07110 IF image caption; A07110-1 applications, dilution data, and image captions). For cross-species work, A07110 lists human, mouse, and rat, while rabbit polyclonal A07110-1 lists human and mouse; validate staining in the chosen species because the supplied images show human tissue only (catalog: reactivity, host, clonality, and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96PP8 (GBP5_HUMAN, Guanylate-binding protein 5).
- Human Protein Atlas. GBP5 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. GBP5 subcellular location (ICC-IF): Highest expression in NB4: 5.7 nTPM.
- Human Protein Atlas. GBP5 antibody validation summary (1 antibodies).
- Clinicopathologic and molecular landscape of villitis of unknown etiology: insights from a large-scale case-control study in PR China. The journal of pathology. Clinical research 2026 — PMC13093737.
- The Proinflammatory Role of Guanylate-Binding Protein 5 in Inflammatory Bowel Diseases. Frontiers in microbiology 2022 — PMC9201962.
- Helicobacter pylori Infection-Related Long Non-Coding RNA Signatures Predict the Prognostic Status for Gastric Cancer Patients. Frontiers in oncology 2021 — PMC8353258.
- GBP5 Identifies Immuno-Hot Tumors and Predicts the Therapeutic Response to Immunotherapy in NSCLC. International journal of general medicine 2023 — PMC10183185.
- PubMed PMID:15175044 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GBP5 (Guanylate-binding protein 5). Expected localisation Cytoplasm in a subset of immune cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A07110 at 1:100–1:500 (standard starting range) (the product's own IHC validation image is shown). Controls (HPA): positive Gallbladder (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A07110); 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 citrate buffer, pH 6.0 , 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 anti-GBP5, 1:100–1:500 starting range (standard); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Sparse positive cells may be missed in sampled fields (HPA tissue IHC); Isoform / epitope: Two isoforms; verify antibody epitope coverage (UniProt); Regulation: IFN-inducible (UniProt). Evidence: UniProt Q96PP8, HPA tissue IHC · Supported, HPA cell-line RNA · Highest expression in NB4: 5.7 nTPM, Datasheet A07110, PMC13093737, PMC9201962. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GBP5/immunohistochemistry-ihc-guide.