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STXBP6 / Syntaxin-binding protein 6 · IHC design guide
Design Immunohistochemistry for STXBP6
Plan chromogenic STXBP6 IHC in paraffin sections using the IHC-validated antibody at 2–5 μg/ml (datasheet: A11292-2). Compare cytoplasmic staining in Purkinje cells and cortical neurons with the listed unstained cell populations (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
STXBP6 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 (HPA tissue IHC); membrane association (UniProt) | |
| Staining pattern | Variable cytoplasmic staining in many cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11292-2) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11292-2) | |
| Caveat | Staining varies by tissue and cell type (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended STXBP6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by two published mouse tumor IHC protocols (PMC7418817; PMC10355827).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A11292-2) |
| Fixation | Image fixative and duration unreported (datasheet A11292-2); 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 A11292-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11292-2) |
| Primary antibody | Rabbit anti-STXBP6, 2-5 μg/ml (datasheet A11292-2) |
| Primary incubation | Overnight at 4 °C (datasheet A11292-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11292-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | STXBP6-positive staining in purkinje cells of cerebellum (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression of varying levels in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A11292-2). The published methods used citrate pH 6.0 or EDTA pH 9.0 (PMC7418817; PMC10355827).
Section 2
What Is the Expected STXBP6 Staining Pattern?
STXBP6 is annotated in the cytoplasm and at membranes, with no transmembrane segment (UniProt Q8NFX7: location and topology). In paraffin section IHC, expect cytoplasmic staining in selected neurons and glandular cells, including cerebellar Purkinje cells and colonic glandular cells at medium intensity (HPA: tissue IHC). HPA rates the tissue pattern Approved, pending external verification (HPA: reliability).
| Cytoplasmic staining in cerebellar Purkinje cells, cortical or hippocampal neurons, or colonic glandular cells. | This matches the reported medium staining in those cell types (HPA: tissue IHC). Assess the named cells and compartment against the local negative control; membrane association is compatible with the UniProt annotation (UniProt Q8NFX7: location). |
| Predominantly nuclear staining, especially when cytoplasmic staining is absent. | A nuclear pattern falls outside the reported cytoplasmic tissue profile and UniProt location annotation (HPA: tissue IHC; UniProt Q8NFX7: location). Check control sections and staining conditions before interpreting it as STXBP6. |
| Strong signal in adipocytes or cardiomyocytes while the expected positive cells are unstained. | Those cell types were reported as not detected (HPA: tissue IHC). Consider nonspecific antibody binding or endogenous chromogenic activity (standard IHC practice); a signal elsewhere is not automatically wrong because HPA describes varying cytoplasmic expression across most tissues (HPA: tissue IHC). |
| Diffuse colour across cells and tissue spaces without a discernible cellular pattern. | This cannot be scored as the reported cytoplasmic pattern (HPA: tissue IHC). Assess background with a no-primary control, then review blocking, washes, antibody concentration and detection chemistry (standard IHC practice). |
| No signal in a cerebellum or colon control section. | The sampled Purkinje or glandular cells should provide a medium-level reference (HPA: tissue IHC). Confirm that those cells are present, then check antibody and detection controls; HPA approval remains pending external verification (HPA: reliability). |
💡Expected STXBP6 appearanceCall a result consistent when the named neurons or glandular cells show discernible, approximately medium cytoplasmic staining (HPA: tissue IHC); isolated nuclear signal or strong signal restricted to HPA not-detected cells warrants control-based review (HPA: tissue IHC; UniProt Q8NFX7: location).
| Tissue and cell selection. | Use a reported medium-staining population as a positive reference, such as Purkinje cells or colonic glandular cells (HPA: tissue IHC). Adipocytes and cardiomyocytes were not detected in the HPA examples (HPA: tissue IHC); evaluate the specified cell type, not the entire organ. |
| Topology and processing. | STXBP6 is annotated as cytoplasmic and membrane-associated without a transmembrane segment, signal peptide or propeptide (UniProt Q8NFX7: location, topology and processing). These annotations guide compartment interpretation; they do not establish an antibody epitope or a target-specific retrieval condition. |
| Isoforms and antibody validation. | UniProt lists three isoforms (UniProt Q8NFX7: isoforms). The supplied HPA records mark HPA003552 and HPA074869 Approved for IHC; neither record reports ICC validation (HPA: antibodies). Epitope coverage across isoforms is not supplied, so do not infer isoform-specific staining. |
| IF/ICC: what pattern is established? | HPA supplies no main ICC/IF location and no cell-line ICC/IF images (HPA: subcellular). Cytoplasm and membrane are UniProt annotations, not a demonstrated IF pattern (UniProt Q8NFX7: location). Interpret an IF result with its own controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive control has no visible staining. | The sampled section may lack the relevant cells, or the IHC detection workflow may have failed (standard IHC practice). | Locate Purkinje cells or colonic glandular cells, reported at medium intensity (HPA: tissue IHC); verify the antibody, detection reagents and control slide (standard IHC practice). |
| Signal is weak in a reported positive population. | Low signal may reflect staining conditions; the supplied sources do not establish STXBP6-specific fixation or retrieval sensitivity. | Check the section and detection controls, then optimize retrieval and antibody concentration within the chosen IHC workflow (standard IHC practice). Do not assume a target-specific retrieval requirement. |
| Brown signal persists when primary antibody is omitted. | Primary-independent signal points to endogenous detection activity or reagent background (standard chromogenic IHC practice). | Review the detection system, apply the relevant endogenous-activity block and compare with the no-primary control (standard chromogenic IHC practice). |
| Diffuse staining obscures cell boundaries. | High antibody concentration, insufficient blocking or incomplete washes can raise background (standard IHC practice). | Optimize dilution, blocking and washing against the positive section and no-primary control (standard IHC practice); score only a resolved cellular pattern. |
| Nuclear signal dominates the slide. | That compartment disagrees with the reported cytoplasmic tissue pattern (HPA: tissue IHC) and UniProt location annotation (UniProt Q8NFX7: location). | Compare positive and no-primary controls, then review counterstain and detection conditions (standard IHC practice) before assigning STXBP6 positivity. |
| A reportedly not-detected cell type stains strongly. | The observation conflicts with that HPA cell-type example (HPA: tissue IHC); nonspecific binding or endogenous activity is possible (standard IHC practice). | Confirm cell identity, examine the no-primary control and compare a reported positive population (standard IHC practice; HPA: tissue IHC). Treat the discrepancy as unresolved until controls support it. |
Experiment evidence
IHC & IF images showing STXBP6 staining
A11292-2 · IHCIHC analysis of STXBP6 using anti-STXBP6 antibody (A11292-2). STXBP6 was detected in a paraffin-embedded section of mouse brain tissue. Heat
A11292-2 · IHCIHC analysis of STXBP6 using anti-STXBP6 antibody (A11292-2). STXBP6 was detected in a paraffin-embedded section of mouse brain tissue. Heat
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168952-STXBP6/tissue/cerebral+cortex#imid_1383
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168952-STXBP6/tissue/cerebral+cortex#imid_20726480
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168952-STXBP6/tissue/colon#imid_20726398
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168952-STXBP6/tissue/liver#imid_20726415
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for STXBP6 IHC & IF
🧪Run cerebellum first and look for staining in Purkinje cells (HPA: Medium in cerebellar Purkinje cells); use esophageal squamous epithelium as the negative tissue (HPA: Not detected in esophageal squamous epithelial cells). On the cerebellar section, use nearby non-Purkinje cells with no signal above background as internal negative comparators, without assuming all neighboring cells lack STXBP6 (HPA: Purkinje-cell staining; standard IHC practice).
Positive control tissue: Cerebellum (Purkinje 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 STXBP6; derive a cell-line control from the positive tissue's cell type (Purkinje cells) and confirm it by RNA or western blot first.
Technical controls: Include a secondary-only control with the primary omitted and a concentration-matched rabbit control IgG matched to the primary’s isotype and clonality where applicable (caption: rabbit primary; standard IHC practice). Confirm specificity with STXBP6 knockout material or immunizing-peptide competition if available, and quench endogenous peroxidase for cerebellar HRP/DAB staining (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU paraffin-section caption does not state the fixative (caption: A11292-2, fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required (caption: A11292-2). The supplied evidence does not establish that frozen sections or IF are easier; if evaluating cerebellar IF, check neuronal autofluorescence (standard IF practice).
HPA tissue IHC evidence for STXBP6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced STXBP6 IHC Tips
Troubleshoot STXBP6 staining in paraffin sections by checking retrieval, compartment, cell type, and assay controls before assigning biological meaning.
How should I retrieve STXBP6 in paraffin sections when staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A11292-2). The catalog antibody stained a paraffin-embedded mouse brain section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A11292-2). If staining is weak, compare retrieval heating and cooling conditions on adjacent sections while keeping antibody concentration and detection constant. Use a tissue region expected to stain, such as cerebellar Purkinje cells, and review background alongside signal (HPA: Medium in Purkinje cells). Record the heating method and duration because the caption specifies the buffer and pH but gives no retrieval time (datasheet A11292-2).
Could fixation explain weak or uneven STXBP6 staining?
The selected image documents a paraffin-embedded mouse brain section but does not state its fixative (datasheet A11292-2). Target-specific STXBP6 sensitivity to fixation is therefore unknown; do not infer it from tissue staining patterns or protein topology. When comparing specimens, document fixation method and duration, processing history, and section age before changing the antibody conditions. Examine adjacent sections processed together, and compare preserved tissue architecture with the distribution of DAB signal. If an experimental fixation change improves staining, confirm the result across matched sections with the same EDTA pH 8.0 retrieval and 2 μg/ml primary incubation (datasheet A11292-2).
Where should convincing STXBP6 staining appear within positive cells?
Expect predominantly cytoplasmic staining in tissue IHC, while allowing a membrane-associated component (HPA: cytoplasmic expression in most tissues; UniProt Q8NFX7: cytoplasm and membrane). STXBP6 has no transmembrane segment, so a sharp membrane outline alone deserves closer scrutiny rather than automatic assignment to the target (UniProt Q8NFX7 topology). Compare the stained cells with surrounding tissue architecture and a serial section before calling a compartment-specific pattern. Cerebellar Purkinje cells and cerebral cortical neuronal cells provide documented medium-level tissue staining references (HPA: cerebellum and cerebral cortex). Persistent nuclear-only signal should prompt review of primary-omission controls, counterstain, and detection background before interpretation.
How can epitope uncertainty affect interpretation across STXBP6 isoforms?
STXBP6 has 3 listed isoforms, but the supplied caption does not identify the antibody epitope or establish isoform coverage (UniProt Q8NFX7 isoforms; datasheet A11292-2). Request the immunogen or mapped epitope information before interpreting differences between cell populations as isoform-specific. The v-SNARE homology region spans residues 151–210; whether this antibody recognizes that region is unspecified (UniProt Q8NFX7 domains; datasheet A11292-2). Compare staining with an independently characterized epitope only if its recognition and IHC performance are documented. Until then, report the observed cellular staining as STXBP6 immunoreactivity, with antibody-dependent epitope coverage unresolved.
How should I assess STXBP6 by IF alongside cell-type markers?
Treat IF as a separate optimization: the supplied antibody image establishes paraffin-section chromogenic IHC, and HPA lists no ICC/IF images for STXBP6 (datasheet A11292-2; HPA subcellular record). Multiplex with a neuronal marker in brain tissue or an epithelial marker in glandular tissue to check the expected cell population (HPA: medium staining in neuronal and glandular cells). Choose spectrally separated fluorophores, favoring a far-red channel when specimen autofluorescence affects shorter wavelengths. Optimize permeabilisation against the antibody’s mapped epitope: STXBP6 is reported in cytoplasm and membrane, but has no transmembrane segment or supplied epitope orientation (UniProt Q8NFX7; datasheet A11292-2). Include single-color and primary-omission controls before interpreting overlap.
What should I adjust when DAB background obscures STXBP6?
First inspect primary-omission sections and the distribution of precipitate to separate detection background from cellular staining. The illustrated method used 10% goat serum blocking, 2 μg/ml primary overnight at 4°C, and DAB detection (datasheet A11292-2). For a chromogenic workflow, check peroxidase blocking, washing, secondary-antibody specificity, and DAB development time on adjacent sections. Reduce development or primary exposure incrementally if diffuse color obscures cells, while retaining an expected positive region. Compare candidate signal with documented medium staining in Purkinje or cortical neuronal cells and absent staining in adipocytes (HPA: cerebellum, cerebral cortex, adipose tissue).
How can I quantify STXBP6 IHC across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, because reported staining varies across tissues and cell types (HPA: tissue IHC profile). For cellular staining, report an H-score from intensity categories and the percentage of positive cells; specify the threshold using the same control sections throughout. For spatial analysis, count positive cells per mm² of viable tissue and report total evaluable cells or area as the denominator. Normalize comparisons to matched regions, section quality, staining batch, and cell composition rather than raw whole-section DAB darkness. Document cytoplasmic scoring separately from any membrane-associated signal (HPA: cytoplasmic expression; UniProt Q8NFX7: cytoplasm and membrane).
How do I distinguish true STXBP6 signal from staining artefacts?
Give most weight to reproducible cytoplasmic staining in an expected cell population, such as Purkinje or cortical neuronal cells (HPA: cytoplasmic expression; HPA: medium in those cells). STXBP6 is also associated with membranes, but has no transmembrane segment, so isolated nuclear or sharply outlined membrane staining needs additional scrutiny (UniProt Q8NFX7 localisation and topology). Review section edges and necrotic areas for nonspecific DAB accumulation, and compare the primary-omission section for endogenous enzyme or detection signal. Use adipocytes as a documented unstained comparator only when that tissue is included in the experiment (HPA: adipocytes not detected). Interpret any positive result with the antibody epitope and isoform coverage still unresolved (UniProt Q8NFX7 isoforms; datasheet A11292-2).
Boster reagents
Best STXBP6 / Syntaxin-binding protein 6 IHC Antibodies
A11292-2 has IHC images from mouse and rat paraffin brain sections (catalog image captions); its listed reactivity includes human, mouse and rat (catalog applications/reactivity). No IF image is supplied (catalog images).
Real IHC data
Anti-STXBP6 Antibody ®
Cat # A11292-2
A11292-2 is listed for IHC and reacts with human, mouse and rat (catalog applications/reactivity). Its IHC images document mouse and rat paraffin brain sections; the fixative is unreported (A11292-2 image captions).
Which to pick: Choose A11292-2 for paraffin-section IHC: its rabbit antibody was used at 2 μg/ml after EDTA pH 8.0 heat retrieval in mouse and rat brain sections (A11292-2 image captions). For cross-species work, its listed reactivity covers human, mouse and rat, while the supplied tissue images document mouse and rat only (catalog reactivity; A11292-2 image captions). There is no IF/ICC-validated SKU in this payload (catalog applications/images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NFX7 (STXB6_HUMAN, Syntaxin-binding protein 6).
- Human Protein Atlas. STXBP6 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. STXBP6 subcellular location (ICC-IF): Highest expression in CACO-2: 40.5 nTPM.
- Human Protein Atlas. STXBP6 antibody validation summary (2 antibodies).
- STXBP6, reciprocally regulated with autophagy, reduces triple negative breast cancer aggressiveness. Clinical and Translational Medicine 2020 — PMC7418817.
- STXBP6-driven signaling cascade alters lipid peroxidation in tumor-associated macrophages to facilitate ovarian cancer progression. Redox biology 2026 — PMC13393757.
- Macrophage fusion event as one prerequisite for inorganic nanoparticle-induced antitumor response. Science advances 2023 — PMC10355827.
- PubMed PMID:12145319 — UniProt-cited evidence.
- PubMed PMID:11042152 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for STXBP6 (Syntaxin-binding protein 6). Expected localisation Cytoplasmic staining (HPA tissue IHC); membrane association (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11292-2). Validated IHC antibody: A11292-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A11292-2); 2. Fixation: Image fixative and duration unreported (datasheet A11292-2); 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 A11292-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11292-2); 8. Primary antibody: Rabbit anti-STXBP6, 2-5 μg/ml (datasheet A11292-2); 9. Primary incubation: Overnight at 4 °C (datasheet A11292-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11292-2); Caveat: Staining varies by tissue and cell type (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q8NFX7, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in CACO-2: 40.5 nTPM, Datasheet A11292-2, PMC7418817, PMC10355827. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/STXBP6/immunohistochemistry-ihc-guide.