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STING1 / Stimulator of interferon genes protein · IHC design guide
Design Immunohistochemistry for STING1
Plan chromogenic STING1 IHC in paraffin sections around cytoplasmic tissue staining (HPA tissue IHC) and an ER/perinuclear molecular expectation (UniProt). The guide highlights strong staining in lung macrophages (HPA tissue IHC) and the catalog antibody’s 2–5 μg/ml IHC range (datasheet A01871-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
STING1 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 tissue staining (HPA tissue IHC); ER/perinuclear location (UniProt) | |
| Staining pattern | Cytoplasmic staining in respiratory, fallopian and lymphoid cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01871-3) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Lung macrophages may dominate mixed-cell staining (HPA tissue IHC) | |
| Regulation | cGAMP triggers ER-to-ERGIC trafficking (UniProt) | |
| Isoform / epitope | No annotated isoforms; residues 135–379 face cytoplasm (UniProt) |
Section 1
Recommended STING1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A01871-3) is followed by published STING1 IHC methods for cholangiocarcinoma (PMC10468663) and papillary renal neoplasms (PMC11573472).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01871-3) |
| Fixation | Image fixative and duration unreported (datasheet A01871-3); 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 A01871-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01871-3) |
| Primary antibody | Rabbit anti-STING1, 2-5 μg/ml (datasheet A01871-3) |
| Primary incubation | Overnight at 4 °C (datasheet A01871-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01871-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | STING1-positive staining in basal cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues including cells in respiratory tract, fallopian tube and cells in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A01871-3); consider EDTA pH 9.0 if optimizing against the published cholangiocarcinoma method (PMC10468663).
Section 2
What Is the Expected STING1 Staining Pattern?
STING1 is a four-pass membrane protein with a large cytoplasmic C-terminal region; inactive STING1 resides at the ER and can move to the ER–Golgi intermediate compartment after activation (UniProt Q86WV6 topology and subcellular location). In tissue IHC, expect cytoplasmic staining in selected cells, especially lung macrophages and bronchial basal cells (HPA: High). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Cytoplasmic staining is strong in lung macrophages or bronchial basal cells, while neighboring cells differ in intensity (HPA: High in these cell types). | This fits the reported cell-specific tissue pattern. Judge signal in the named cells rather than treating every cell in a positive tissue as a positive control (HPA: tissue IHC). The cytoplasmic appearance also fits STING1 membrane topology and ER localization (UniProt Q86WV6). |
| The dominant signal is nuclear-only, with little cytoplasmic staining in otherwise recognizable positive cells. | This is unexpected for tissue IHC, which HPA describes as cytoplasmic (HPA: tissue IHC). It warrants an artefact check, but location alone cannot prove artefact: HPA reports nucleoplasmic ICC-IF staining with uncertain support (HPA: subcellular). |
| Strong staining appears in a cell population reported as not detected, such as skeletal-muscle myocytes or breast adipocytes (HPA: Not detected). | Assess possible cross-reactivity or endogenous detection activity before calling this STING1. These HPA observations apply to the specified cells, not every cell in those tissues (HPA: tissue IHC). Compare the suspicious signal with a known-positive cell population and a control lacking primary antibody (general IHC practice). |
| Broad, diffuse color covers tissue and spaces between cells, obscuring cellular boundaries. | That distribution does not resolve the cytoplasmic, cell-specific pattern reported by HPA (HPA: tissue IHC). Background from detection reagents or incomplete blocking is a general IHC possibility; the image alone does not identify its source. Review a control lacking primary antibody before scoring weak cells (general IHC practice). |
| No cytoplasmic signal is visible in lung macrophages or bronchial basal cells (HPA: High). | A negative result in these reported high-staining cells makes the run difficult to interpret (HPA: tissue IHC). Check that the relevant cells are present, then review antibody application and detection controls (general IHC practice). HPA tissue staining does not establish a STING1-specific fixation or retrieval failure. |
💡Expected STING1 appearanceCall the result positive when recognizable lung macrophages or bronchial basal cells show high cytoplasmic staining (HPA: tissue IHC); nuclear-only staining or uniform color across unrelated cells calls for control review, not automatic STING1 scoring (HPA: tissue IHC; general IHC practice).
| Topology and compartment | STING1 has 4 transmembrane segments and a cytoplasmic region spanning residues 135–379 (UniProt Q86WV6 topology). Its ER and related membrane locations make a cytoplasmic tissue pattern plausible (UniProt Q86WV6 subcellular location; HPA: tissue IHC). The supplied record does not identify the catalog antibody's epitope. |
| Activation state | Inactive STING1 localizes to the ER; after cyclic-dinucleotide activation it moves toward the ER–Golgi intermediate compartment (UniProt Q86WV6 subcellular location). A shift in intracellular distribution can therefore be biologically plausible. Routine tissue staining alone does not establish activation state or prove that trafficking occurred. |
| Tissue and antibody evidence | HPA reports High staining in lung macrophages, bronchial and nasopharyngeal basal cells, fallopian-tube ciliated cells, and tonsil squamous epithelial cells (HPA: tissue IHC). Its tissue reliability is Enhanced, with medium staining–RNA consistency (HPA: tissue IHC). HPA038116 and HPA038534 have Enhanced IHC validation (HPA: antibodies); this does not validate every antibody. |
| IF/ICC: what localization is expected? | HPA reports mainly cytosolic ICC-IF localization and additionally supported vesicle staining (HPA: subcellular). It labels nucleoplasm, primary cilium and basal body locations uncertain (HPA: subcellular). Use those observations to interpret IF images; the IHC tissue pattern remains the reference for this paraffin-section guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive lung macrophages are unstained, and other expected positive cells are also blank (HPA: High in lung macrophages). | The cells may be absent from the examined field, or the primary-antibody or detection step may have failed (general IHC practice). The supplied sources do not identify a STING1-specific fixation effect. | Confirm cell identity in the section, inspect a positive control, and verify that the primary antibody and chromogenic detection steps were applied (general IHC practice). |
| A positive tissue shows weak staining, making cell boundaries and localization hard to judge. | Low signal or an unsuitable staining run can prevent a clear comparison with HPA's cytoplasmic pattern; HPA intensity is cell-specific (HPA: tissue IHC). The payload gives no catalog dilution or retrieval condition. | Compare the named positive cells with a run control; review the antibody's documented IHC conditions, including retrieval and dilution, before changing them (general IHC practice). Do not infer a STING1-specific retrieval requirement from HPA. |
| Color is widespread even where the primary antibody was omitted. | A control lacking primary antibody can reveal signal from detection chemistry or tissue-associated endogenous activity (general IHC practice). That signal cannot be attributed to STING1 from appearance alone. | Check the detection-system controls and apply the appropriate endogenous-activity blocking step for that system (general IHC practice). Reassess cellular localization after background is controlled. |
| Strong nuclear-only staining dominates an IHC section. | The pattern conflicts with HPA's reported cytoplasmic tissue staining (HPA: tissue IHC). HPA's uncertain nucleoplasmic ICC-IF observation does not by itself establish nuclear-only paraffin IHC as the expected result (HPA: subcellular). | Review the control lacking primary antibody and compare cytoplasmic staining in named positive cells; seek confirmation with an independently validated IHC antibody if the interpretation matters (general IHC practice; HPA: antibodies). |
| Skeletal-muscle myocytes or breast adipocytes stain as strongly as the selected positive cells (HPA: Not detected in those cell types). | Cross-reactivity or nonspecific detection is possible (general IHC practice). HPA's negative calls concern the listed cell types, so staining elsewhere in the same tissue is a separate observation (HPA: tissue IHC). | Verify cell identity and compare a control lacking primary antibody, then review antibody specificity before scoring the unexpected cells as STING1-positive (general IHC practice). |
| A section mixes high, medium and absent staining across distinct cell populations. | HPA reports cell-level differences: lung macrophages are High, appendix glandular cells Medium, and adrenal gland glandular cells Not detected (HPA: tissue IHC). Such variation alone does not mean the run failed. | Record intensity and cytoplasmic localization by cell type. Compare each cell population with its own HPA reference and interpret borderline staining alongside controls (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing STING1 staining
A01871-3 · IHCIHC analysis of TMEM173/STING using anti-TMEM173/STING antibody (A01871-3). TMEM173/STING was detected in a paraffin-embedded section of hum
A01871-3 · IFIF analysis of TMEM173/STING using anti-TMEM173/STING antibody (A01871-3) and anti-Beta Tubulin antibody (M01857-3). TMEM173/STING was detec
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184584-STING1/tissue/cerebral+cortex#imid_10021936
HPA · BronchusHuman Protein Atlas IHC, Bronchus. Source: https://www.proteinatlas.org/ENSG00000184584-STING1/tissue/bronchus#imid_10021909
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184584-STING1/tissue/colon#imid_10010838
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184584-STING1/tissue/liver#imid_10010831
![HPA ICC-IF BJ [Human fibroblast] — STING1 (HPA038534)](https://images.proteinatlas.org/38534/1658_C1_2_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA038534; green = STING1, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — STING1 (HPA038534)](https://images.proteinatlas.org/38534/1658_C1_3_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA038534; green = STING1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for STING1 IHC & IF
🧪Run lung first and look for staining in its macrophages (HPA: High in lung macrophages). Run adrenal gland as a negative tissue, assessing its glandular cells (HPA: Not detected in adrenal gland glandular cells); on the lung slide, use neighboring cells that lack specific staining as an internal background reference without assuming every nonmacrophage cell is STING1-negative (HPA: High in lung macrophages).
Positive control tissue: Bronchus (Basal cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show STING1 in BJ [Human fibroblast], EFO-21, U2OS, THP-1, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched control IgG controls; the captioned primary is rabbit-derived (selected-SKU tissue-IHC caption). Use STING1-knockout tissue or cells as a biological specificity control, and quench endogenous peroxidase and inspect lung macrophage pigment before interpreting DAB signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A01871-3 paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption). That caption used heat-mediated EDTA retrieval at pH 8.0, but it does not establish that retrieval is required (selected-SKU tissue-IHC caption). The supplied evidence supports a paraffin IHC starting point without establishing that frozen sections or IF/ICC are easier; assess lung macrophage pigment and endogenous peroxidase when scoring chromogenic staining (selected-SKU tissue-IHC caption; standard IHC practice).
HPA tissue IHC evidence for STING1
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 |
|---|---|---|---|---|
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Tonsil | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced STING1 IHC Tips
Troubleshoot STING1 staining in paraffin sections by checking retrieval, cell type, subcellular pattern and controls before interpreting chromogenic signal.
How should I retrieve STING1 when paraffin sections stain weakly?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A01871-3). The selected paraffin-section example used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody, so reproduce that combination when establishing a baseline (datasheet A01871-3). If staining remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration, detection and development time constant (standard IHC practice). Score specific cellular signal alongside tissue damage and background, using lung macrophages as a positive reference where available (HPA: High in lung macrophages).
Could fixation explain weak or inconsistent STING1 staining?
The selected image documents a paraffin-embedded human lung cancer section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A01871-3: fixative not stated). Record the actual fixative and processing history for each specimen before comparing staining intensity across blocks (standard IHC practice). On matched sections, keep EDTA retrieval at pH 8.0 and the 2 μg/ml primary incubation constant while assessing morphology, specific signal and background (datasheet A01871-3; standard IHC practice). Use a concurrently processed positive control to distinguish a processing problem from variation in the sampled cell population (standard IHC practice).
Which STING1 staining pattern is plausible in chromogenic IHC?
Expect predominantly cytoplasmic or perinuclear staining, with a pattern compatible with membrane-associated intracellular compartments (HPA: cytoplasmic tissue expression; UniProt Q86WV6: ER and perinuclear localisation). Inactive STING1 resides at the ER, while activated protein can move through the ER-Golgi intermediate compartment and Golgi-related membranes (UniProt Q86WV6: subcellular localisation). Interpret apparent diffuse nuclear-only DAB cautiously because the protein has 4 transmembrane segments and a cytoplasmic C-terminal region (UniProt Q86WV6: topology). Compare suspicious staining with adjacent-section controls and cellular morphology before assigning a compartment or activation state (standard IHC practice).
How do epitope position and processing affect STING1 IHC interpretation?
The supplied record lists 0 isoforms and a single 1–379 chain, so an isoform-specific staining explanation is unsupported here (UniProt Q86WV6: isoforms and processing). STING1 has 4 transmembrane segments within residues 18–134 and a cytoplasmic region spanning 135–379 (UniProt Q86WV6: topology). The antibody epitope is not supplied, so check its documented immunogen before attributing weak staining to membrane accessibility or a modified residue (datasheet A01871-3: epitope not supplied; standard IHC practice). Keep retrieval and detection matched across sections when comparing samples with different staining patterns (standard IHC practice).
How should I follow up an IHC pattern with multiplex IF?
Use a separate IF/ICC-validated workflow to test whether STING1 signal overlaps an expected cell-type marker, such as a macrophage marker in lung tissue (HPA: High in lung macrophages; standard IF practice). Include single-stain controls and select spectrally separated fluorophores after checking tissue autofluorescence in each channel (standard IF practice). Set permeabilisation according to the documented antibody epitope: the 135–379 region faces the cytoplasm, while short loops at 35–44 and 107–116 face the lumen (UniProt Q86WV6: topology). The paraffin-section caption does not establish IF fixation or permeabilisation conditions for this antibody (datasheet A01871-3: IHC caption).
What should I change when DAB staining obscures specific STING1 signal?
Check a no-primary control and inspect whether the apparent signal follows tissue edges, damaged areas or broadly stained extracellular material (standard IHC practice). The selected example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked detection system with DAB (datasheet A01871-3). Include an endogenous peroxidase block for chromogenic detection, then adjust primary concentration or DAB development only after verifying retrieval and washes (standard IHC practice). Judge improvement by retained cellular staining in a positive control, such as lung macrophages, alongside reduced no-primary background (HPA: High in lung macrophages; standard IHC practice).
How should I quantify STING1 staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and region before reading slides, since HPA reports High staining in lung macrophages but other tissue compartments vary (HPA: tissue IHC). For cellular intensity, record the percentage at each prespecified intensity grade and calculate an H-score; alternatively report % positive cells within the defined population (standard IHC practice). For infiltrating positive cells, report density per mm² of viable analysed tissue, with the same segmentation and exclusion rules across samples (standard IHC practice). Normalise comparisons to the number of eligible cells or viable area, and record cytoplasmic versus punctate patterns separately (standard IHC practice; UniProt Q86WV6: subcellular localisation).
How can I separate true STING1 positivity from staining artefacts?
A credible result combines cellular staining in a plausible compartment with an appropriate positive control: HPA reports High staining in lung macrophages and cytoplasmic expression across several tissues (HPA: tissue IHC). ER-associated or perinuclear signal is compatible with inactive STING1, while redistribution toward Golgi-related compartments may occur after activation (UniProt Q86WV6: subcellular localisation). Treat isolated nuclear-only staining, section-edge accentuation and necrotic-area deposits as suspect until controls and morphology support them (UniProt Q86WV6: topology; standard IHC practice). Compare a no-primary control and peroxidase-blocked section when brown signal could reflect endogenous enzyme activity rather than specific DAB detection (standard IHC practice).
Boster reagents
Best STING1 / Stimulator of interferon genes protein IHC Antibodies
A01871-3 has IHC data from human lung cancer paraffin sections and IF/ICC data from SIHA cells; the catalog lists human, mouse and rat reactivity (A01871-3 image captions; catalog reactivity).
Real IHC data
Anti-TMEM173/STING Antibody ®
Cat # A01871-3
A01871-3 was demonstrated by IHC on a human lung cancer paraffin section (A01871-3 IHC caption). The same SKU was demonstrated by IF/ICC in SIHA cells; its listed applications include IHC, IF and ICC (A01871-3 IF caption; catalog applications).
Which to pick: Choose A01871-3 for tissue IHC because its own caption documents staining of a paraffin section with 2 μg/ml primary antibody after EDTA retrieval (A01871-3 IHC caption). Choose the same SKU for IF/ICC because its IF caption documents SIHA cell staining at 5 μg/ml (A01871-3 IF caption). For mouse or rat samples, the catalog lists reactivity, but the supplied IHC and IF examples are human; the tissue fixative is unreported (catalog reactivity; A01871-3 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q86WV6 (STING_HUMAN, Stimulator of interferon genes protein).
- Human Protein Atlas. STING1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. STING1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm, vesicles, primary cilium and basal body..
- Human Protein Atlas. STING1 antibody validation summary (3 antibodies).
- Proteomic Profiling Identifies Predictive Signatures for Progression Risk in Patients with Advanced-Stage Follicular Lymphoma. Cancers 2024 — PMC11476298.
- SMAD4 regulates the progression of cholangiocarcinoma by modulating the expression of STING1. Journal of cellular and molecular medicine 2023 — PMC10468663.
- Histopathologic classification and immunohistochemical features of papillary renal neoplasm with potential therapeutic targets. Journal of pathology and translational medicine 2024 — PMC11573472.
- Stimulator of interferon genes (STING) expression in the enteric nervous system and contributions of glial STING in disease. Neurogastroenterology and motility 2023 — PMC10266835.
- PubMed PMID:18818105 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for STING1 (Stimulator of interferon genes protein). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER/perinuclear location (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01871-3). Validated IHC antibody: A01871-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Basal cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01871-3); 2. Fixation: Image fixative and duration unreported (datasheet A01871-3); 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 A01871-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01871-3); 8. Primary antibody: Rabbit anti-STING1, 2-5 μg/ml (datasheet A01871-3); 9. Primary incubation: Overnight at 4 °C (datasheet A01871-3). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Lung macrophages may dominate mixed-cell staining (HPA tissue IHC); Isoform / epitope: No annotated isoforms; residues 135–379 face cytoplasm (UniProt); Regulation: cGAMP triggers ER-to-ERGIC trafficking (UniProt). Evidence: UniProt Q86WV6, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A01871-3, PMC10468663, PMC11573472. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/STING1/immunohistochemistry-ihc-guide.