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GBP1 / Guanylate-binding protein 1 · IHC design guide
Design Immunohistochemistry for GBP1
Plan GBP1 staining in paraffin sections using an IHC-validated antibody. This guide covers fixation, controls and interpretation of the cytoplasmic staining reported in a few tissues, most abundant in immune cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
GBP1 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 a few tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal, most abundant in immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03067-1) | |
| Positive control | Appendix+2 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Immune-cell staining is unannotated; RNA concordance is low (HPA tissue IHC) | |
| Regulation | Interferon inducible (UniProt) | |
| Isoform / epitope | No listed isoforms; residues 590–592 are processed (UniProt) |
Section 1
Recommended GBP1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A03067-1) with three published GBP1 IHC protocols (PMC10710186; PMC11865198; PMC5978929).
| Sample | Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A03067-1) |
| Fixation | Image fixative and duration unreported (datasheet A03067-1); 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 A03067-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03067-1) |
| Primary antibody | Rabbit anti-GBP1, 2-5 μg/ml (datasheet A03067-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03067-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03067-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | GBP1-positive staining in lymphoid tissue of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in a few tissues, most abundant in immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A03067-1). Citrate at pH 6.0 was reported for salivary gland sections (PMC10710186).
Section 2
What Is the Expected GBP1 Staining Pattern?
GBP1 is chiefly cytosolic and can associate with cytoplasmic vesicle, Golgi, and cell membranes; it has no transmembrane segment (UniProt P32455 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic staining in selected cells, including immune cells and some glandular cells (HPA tissue IHC). Interpret intensity cautiously: HPA rates tissue staining “Approved” but reports low consistency with RNA expression and does not annotate immune-cell staining individually (HPA tissue IHC).
| Cytoplasmic staining in appendix lymphoid tissue or small-intestinal or thyroid glandular cells, with a medium signal (HPA tissue IHC). | This matches the recorded tissue and cell patterns. Score the stained cell population and compartment together; a positive gland does not imply that every nearby cell should stain (HPA tissue IHC). |
| Predominantly nuclear staining, with little corresponding cytoplasmic signal. | Treat this as a localisation mismatch requiring investigation: UniProt lists cytosolic and membrane-associated locations, but no nuclear location (UniProt P32455 subcellular location). Review morphology, counterstain, and controls before assigning it to GBP1 (general IHC practice). |
| Strong staining in an HPA-listed negative cell population, such as adrenal glandular cells or bone-marrow hematopoietic cells (HPA tissue IHC). | Consider nonspecific binding or endogenous chromogen-detection activity, especially if a reagent-omission control also develops colour (general IHC practice). HPA's “not detected” result is cell-type specific and does not prove every cell in that tissue is negative (HPA tissue IHC). |
| Diffuse colour across cells and extracellular areas, obscuring cell boundaries. | This distribution is difficult to score as GBP1 because the documented tissue pattern is cytoplasmic in selected cells (HPA tissue IHC). Check background controls and staining development before interpreting weak cell-associated colour (general IHC practice). |
| No convincing signal in appendix lymphoid tissue or the listed positive glandular cells (HPA tissue IHC). | The expected comparator is medium staining in those specified cells, not uniform staining of the whole section (HPA tissue IHC). Assess whether those cells are present and whether the run controls worked before calling the specimen negative (general IHC practice). |
💡Expected GBP1 appearanceCall a result convincing when selected immune or glandular cells show discernible cytoplasmic signal, with medium staining in the listed positive populations (HPA tissue IHC); predominant nuclear colour or similarly strong staining in listed negative cell populations warrants a control check (UniProt P32455 subcellular location; HPA tissue IHC).
| Tissue and cell context | HPA records medium staining in appendix lymphoid tissue and small-intestinal and thyroid glandular cells; lung macrophages are low (HPA tissue IHC). Choose and score controls by the stated cell population, not by tissue name alone (general IHC practice). |
| Confidence in the tissue pattern | HPA rates the tissue IHC pattern “Approved,” reports low agreement with RNA, and leaves immune-cell staining unannotated in its summary (HPA tissue IHC). Resolve ambiguous cells against morphology and controls rather than treating the summary as a cell-by-cell map (general IHC practice). |
| GBP1 distribution | GBP1 has no transmembrane segment and is listed in cytosol and at several membranes (UniProt P32455 topology and subcellular location). Infection-associated recruitment to pathogen vacuoles or escaped bacteria is documented; do not require that specialised pattern in routine tissue (UniProt P32455 subcellular location). |
| Processing and epitope uncertainty | UniProt lists a 1–589 chain, a 590–592 propeptide, and modifications at residues 156, 589, and 590 (UniProt P32455 processing and modified residues). No antibody epitope is supplied, so these annotations alone cannot predict staining loss or a retrieval requirement. |
| IF/ICC Q: Can this IHC pattern be used as an IF control? | A: Use the UniProt locations as biological context, but HPA provides no ICC-IF images or main-location assignment, and the listed antibody has no ICC validation entry (UniProt P32455 subcellular location; HPA subcellular and antibody records). This page cannot establish an IF staining pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Listed positive cells are unstained while the run control also fails. | A run-level staining or detection problem is plausible (general IHC practice). The HPA comparator is medium staining in specified appendix, small-intestinal, or thyroid cell populations (HPA tissue IHC). | Confirm the expected cells are present, then review reagent application, detection, chromogen development, and the laboratory's antigen-retrieval conditions with a working control section (general IHC practice). |
| Listed positive cells are unstained but the run control works. | Sampling or cell identification may explain the discrepancy; the HPA result applies to named cell populations and has low RNA–staining consistency (HPA tissue IHC). | Recheck morphology and the cells actually scored; compare another listed positive population before concluding that this specimen lacks GBP1 (HPA tissue IHC; general IHC practice). |
| Nuclear colour dominates the section. | The observed compartment conflicts with the listed cytosolic and membrane-associated GBP1 locations (UniProt P32455 subcellular location). Counterstain overlap or nonspecific detection can complicate interpretation (general IHC practice). | Inspect an omission control and separate chromogen from counterstain by microscopy; score GBP1 only where cell-associated signal can be assigned confidently (general IHC practice). |
| Colour appears in listed negative cells, including adrenal glandular cells (HPA tissue IHC). | Nonspecific antibody binding or endogenous detection activity is possible (general IHC practice); HPA reports those particular cells as not detected (HPA tissue IHC). | Compare a primary-antibody omission control, review blocking and detection reagents, and assess whether the signal follows identifiable cells rather than tissue deposits (general IHC practice). |
| Diffuse background hides a weak cytoplasmic pattern. | Excess detection development, inadequate washing, or nonspecific binding can raise background in chromogenic IHC (general IHC practice). | Use the omission control to locate background, review washing and chromogen-development time, and compare a listed positive cell population in the same run (general IHC practice; HPA tissue IHC). |
| Staining is present in lung macrophages but weaker than in appendix lymphoid tissue. | This can fit the reported low lung-macrophage and medium appendix-lymphoid staining levels (HPA tissue IHC). | Identify macrophages by morphology and compare like cell populations under the same scoring criteria; do not label the weaker lung signal a failed run solely from its intensity (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing GBP1 staining
A03067-1 · IHCIHC analysis of GBP1 using anti-GBP1 antibody (A03067-1). GBP1 was detected in a paraffin-embedded section of human ovarian cancer tissue. H
A03067-1 · IHCIHC analysis of GBP1 using anti-GBP1 antibody (A03067-1). GBP1 was detected in a paraffin-embedded section of mouse heart tissue. Heat media
A03067-1 · IFIF analysis of GBP1 using anti-GBP1 antibody (A03067-1). GBP1 was detected in an immunocytochemical section of SiHa cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117228-GBP1/tissue/cerebral+cortex#imid_17892123
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117228-GBP1/tissue/colon#imid_17892033
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117228-GBP1/tissue/liver#imid_17892034
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000117228-GBP1/tissue/kidney#imid_17892035
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for GBP1 IHC & IF
🧪Run appendix first: lymphoid tissue should stain for GBP1 (HPA: Medium in appendix lymphoid tissue). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); on the appendix slide, assess non-lymphoid cells as internal negatives and expect no specific signal above control background.
Positive control tissue: Appendix (Lymphoid tissue, HPA Medium)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for GBP1; derive a cell-line control from the positive tissue's cell type (Lymphoid tissue) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) slide, a species- and isotype-matched rabbit IgG control (caption: rabbit primary), and GBP1-knockout material as a biological negative. Block endogenous peroxidase in appendix sections before chromogenic detection and compare any residual staining with the no-primary slide (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03067-1 paraffin-section caption does not state the fixative. That caption reports heat-mediated retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required under other conditions. The supplied evidence does not establish that frozen sections or IF are easier; for appendix IHC, endogenous peroxidase in blood or inflammatory cells can complicate interpretation (standard IHC practice).
HPA tissue IHC evidence for GBP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Antibody staining in immune cells not annotated, view images.). 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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | Medium | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GBP1 IHC Tips
Troubleshoot GBP1 staining in paraffin sections by checking retrieval, cellular localisation and controls before comparing staining across samples.
What retrieval should I try first if GBP1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03067-1). The selected antibody detected GBP1 in a human ovarian cancer section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A03067-1). If staining remains weak, check section adhesion, retrieval temperature and heating consistency before changing conditions (standard IHC practice). Compare any adjusted retrieval condition on adjacent sections with the same controls and detection reagents, since stronger DAB signal alone cannot establish specificity (standard IHC practice). Record the condition used for every scored slide (standard IHC practice).
Can fixation explain weak or uneven GBP1 staining?
The selected tissue caption describes a paraffin section but does not state its fixative, so GBP1-specific fixation sensitivity is unknown (datasheet A03067-1). Check the specimen’s fixation record, section thickness and tissue preservation when staining varies across a slide or between cases (standard IHC practice). Run similarly processed control sections together to distinguish a specimen-processing problem from a staining-run problem (standard IHC practice). Do not assign a GBP1-specific fixation effect based on tissue staining patterns or protein topology; neither supplies a fixation comparison (HPA tissue IHC; UniProt P32455 topology). Document processing differences before interpreting a negative result (standard IHC practice).
Where should GBP1 signal appear in a paraffin section?
Assess cytoplasmic signal first: tissue staining is described as cytoplasmic and most abundant in immune cells (HPA tissue IHC). GBP1 is also annotated in cytosol and at vesicle, Golgi and cell membranes, with recruitment to pathogen-containing vacuoles or exposed bacteria under relevant conditions (UniProt P32455 subcellular location). Its Golgi localisation depends on isoprenylation and another IFNG-induced factor, so punctate or membrane-associated signal needs cellular context (UniProt P32455 subcellular location). Score the stained cell type and subcellular pattern separately from intensity (standard IHC practice). Predominantly nuclear DAB without credible cytoplasmic staining warrants review of controls and morphology (HPA tissue IHC; standard IHC practice).
How should epitope uncertainty affect interpretation of GBP1 staining?
The supplied record lists 0 isoforms and no transmembrane segment, but gives no epitope map for this antibody (UniProt P32455 isoforms and topology; datasheet A03067-1). GBP1 has a GTPase domain at residues 35–278 and reported modifications at residues 156, 589 and 590 (UniProt P32455 domains and modified residues). Without epitope information, do not attribute a staining difference to a particular modification or protein terminus (standard IHC interpretation). Compare retrieval conditions and a suitable independent specificity control if signal changes unexpectedly (standard IHC practice). Report the antibody identifier and retrieval method so others can evaluate the result (standard IHC practice).
How can IF help clarify an ambiguous GBP1 IHC pattern?
Use IF as a separate localisation check when chromogenic GBP1 staining leaves cell identity or compartment uncertain (standard IHC and IF practice). Multiplex GBP1 with a validated marker for the cell type being assessed, since the tissue profile describes GBP1 as most abundant in immune cells (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence and include single-label controls to assess spectral spillover (standard IF practice). For a cytosolic epitope, use permeabilisation compatible with intracellular access; for a proposed membrane-facing epitope, first establish which side the antibody recognises (UniProt P32455 topology; standard IF practice). The selected caption establishes paraffin-section IHC conditions, not IF performance (datasheet A03067-1).
What should I check when GBP1 DAB staining is diffuse?
First compare the suspect section with a no-primary control and inspect whether colour tracks tissue edges, damaged regions or pigment rather than intact cells (standard IHC practice). Review peroxidase blocking, secondary-antibody background, wash quality and DAB development time using the same tissue-processing batch (standard IHC practice). The selected example used a goat-serum block, an anti-rabbit peroxidase secondary and DAB detection; these are reported conditions, not proof that every brown deposit is GBP1 (datasheet A03067-1). Interpret diffuse signal against the expected cytoplasmic profile and the local cell morphology (HPA tissue IHC; standard IHC practice). Adjust detection only after controls identify the background source (standard IHC practice).
How should I quantify GBP1 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and cell population before measuring GBP1, because the reported tissue profile is cytoplasmic and enriched in immune cells (HPA tissue IHC). For cell-based scoring, record the percentage of positive cells and an H-score from staining intensity and positive-cell fractions; for infiltrates, consider GBP1-positive cells per mm² (standard IHC practice). Normalise cell counts to viable tissue area or to the number of eligible cells in the defined compartment, and exclude folds and necrosis consistently (standard IHC practice). Keep retrieval, exposure to chromogen and scoring thresholds consistent across compared sections (standard IHC practice). Report cell-type-specific results alongside whole-section summaries (standard IHC practice).
When is GBP1 staining convincing rather than artefactual?
A convincing result follows intact cell boundaries and shows cytoplasmic staining in an identifiable population, with appropriate control behaviour (HPA tissue IHC; standard IHC practice). GBP1 can also associate with vesicle, Golgi or cell membranes, so assess those patterns alongside morphology and experimental context (UniProt P32455 subcellular location). Question isolated nuclear colour, edge-restricted deposits, necrotic staining or signal persisting in a no-primary control; review endogenous peroxidase blocking when DAB background is suspected (HPA tissue IHC; standard IHC practice). The HPA tissue entry is rated Approved but reports low consistency between antibody staining and RNA expression data (HPA tissue IHC). Treat an unexpected positive or negative as provisional until supported by controls and context (standard IHC practice).
Boster reagents
Best GBP1 / Guanylate-binding protein 1 IHC Antibodies
A03067-1 has IHC images from paraffin-embedded human ovarian cancer and mouse and rat heart tissue, plus IF imaging in SiHa cells (catalog image captions).
Real IHC data
Anti-GBP1 Antibody ®
Cat # A03067-1
A03067-1 has pictured IHC in paraffin-embedded human ovarian cancer and mouse and rat heart tissue, and pictured IF in SiHa cells (catalog image captions). M03067-1 lists human IHC and ICC/IF applications, but has no supplied IHC or IF image captions (catalog applications; image captions).
Which to pick: Choose A03067-1 for tissue IHC in paraffin sections: its own captions show human ovarian cancer and mouse and rat heart tissue; the fixative is unreported (A03067-1 IHC image captions). Choose A03067-1 for IF/ICC when a pictured example matters: its caption shows IF in SiHa cells, while M03067-1 lists ICC/IF without a supplied image caption (catalog IF image captions; applications). For cross-species work, A03067-1 lists human, mouse and rat reactivity; M03067-1 is a rabbit monoclonal listing human reactivity only (catalog reactivity; host; clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P32455 (GBP1_HUMAN, Guanylate-binding protein 1).
- Human Protein Atlas. GBP1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. GBP1 subcellular location (ICC-IF): Highest expression in GAMG: 67.1 nTPM.
- Human Protein Atlas. GBP1 antibody validation summary (1 antibodies).
- Guanine nucleotide-binding protein 1 is one of the key molecules contributing to cancer cell radioresistance. Cancer science 2014 — PMC4462352.
- Identification of Key Genes for Pyroptosis-Induced Salivary Gland Inflammation in Sjogren's Syndrome Based on Microarray Data and Immunohistochemistry Analysis. Journal of inflammation research 2023 — PMC10710186.
- Role of guanylate-binding protein 1 in the proliferation of invasive lung adenocarcinoma cells. Frontiers in oncology 2025 — PMC11865198.
- T lymphocytes facilitate brain metastasis of breast cancer by inducing Guanylate-Binding Protein 1 expression. Acta neuropathologica 2018 — PMC5978929.
- PubMed PMID:1715024 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for GBP1 (Guanylate-binding protein 1). Expected localisation Cytoplasmic staining in a few tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03067-1). Validated IHC antibody: A03067-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A03067-1); 2. Fixation: Image fixative and duration unreported (datasheet A03067-1); 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 A03067-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03067-1); 8. Primary antibody: Rabbit anti-GBP1, 2-5 μg/ml (datasheet A03067-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03067-1). Watch for: Fixation: Keep section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Immune-cell staining is unannotated; RNA concordance is low (HPA tissue IHC); Isoform / epitope: No listed isoforms; residues 590–592 are processed (UniProt); Regulation: Interferon inducible (UniProt). Evidence: UniProt P32455, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in GAMG: 67.1 nTPM, Datasheet A03067-1, PMC10710186, PMC11865198. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/GBP1/immunohistochemistry-ihc-guide.