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S100A14 / Protein S100-A14 · IHC design guide
Design Immunohistochemistry for S100A14
S100A14 is cytoplasmic (UniProt), while tissue IHC highlights squamous epithelial cells and sometimes membrane staining (HPA tissue IHC). This guide pairs human paraffin-section IHC at 2–5 µg/ml (datasheet A06301-1) with cell-specific controls selected from the tissue profile (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
S100A14 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 (UniProt); membranous in some tissues (HPA tissue IHC) | |
| Staining pattern | Squamous epithelial staining; membranous in some tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06301-1) | |
| Positive control | Appendix+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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Whole-tissue scoring may obscure endocrine-cell staining (HPA tissue IHC) | |
| Regulation | Expression varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | One 1–104 chain, no isoforms; cytoplasmic epitope expected (UniProt) |
Section 1
Recommended S100A14 IHC & IF Protocols
The catalog antibody has a datasheet IHC protocol; four published S100A14 chromogenic IHC protocols provide tissue-specific examples (datasheet A06301-1; PMC4348405; PMC6026614; PMC5550849; PMC4116497).
| Sample | Paraffin-embedded human cervica squamous carcinoma tissue; fixative not specified (datasheet A06301-1) |
| Fixation | Image fixative and duration unreported (datasheet A06301-1); 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 A06301-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06301-1) |
| Primary antibody | Rabbit anti-S100A14, 2-5 µg/ml (datasheet A06301-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06301-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06301-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | S100A14-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Expression in squamous epithelial cells with additional membranous expression in some tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A06301-1). Published protocols used other retrieval conditions with their own antibodies (PMC4348405; PMC6026614).
Section 2
What Is the Expected S100A14 Staining Pattern?
In paraffin-section IHC, expect S100A14 mainly in squamous epithelial cells, with additional membranous staining in some tissues (HPA tissue IHC: Enhanced). Cytoplasmic staining is consistent with its reported location (UniProt Q9HCY8: Cytoplasm). S100A14 has no transmembrane segment, so membrane-associated staining should be judged against the tissue pattern, not treated as evidence of a membrane-spanning protein (UniProt Q9HCY8: topology; HPA tissue IHC: profile).
| Strong staining in esophageal or cervical squamous epithelial cells, with cytoplasmic and possibly membranous signal. | This fits the expected cell type and reported high staining in both tissues (HPA tissue IHC: Esophagus and Cervix, High). Score signal within the squamous cells; the surrounding tissue is not expected to stain uniformly on that basis. |
| Strong staining is confined to scattered endocrine cells in colon or appendix rather than every epithelial cell. | This can be a correct positive result: HPA assigns High staining specifically to endocrine cells in colon and appendix (HPA tissue IHC: Colon and Appendix, High). Check cell identity before calling the section weak or patchy. |
| Signal appears only in extracellular material or stroma, without staining in the expected cells. | That distribution does not match the reported cytoplasmic location or the HPA cell-specific IHC profile (UniProt Q9HCY8: Cytoplasm; HPA tissue IHC: profile). Review morphology and controls before attributing it to S100A14. |
| Broad staining appears in a cell type reported as undetected, or colour develops on the negative control. | Suspect nonspecific binding or endogenous detection activity and compare controls (general IHC practice). HPA reports adipocytes and bronchial respiratory epithelial cells as Not detected; that finding is specific to those cells (HPA tissue IHC). |
| A known-positive squamous epithelium has no visible signal while the tissue and counterstain remain interpretable. | Treat this as a possible assay failure before concluding that S100A14 is absent: esophageal and cervical squamous cells are High in HPA IHC (HPA tissue IHC). Recheck antibody use, detection and controls (general IHC practice). |
💡Expected S100A14 appearanceCall an IHC result positive when the expected epithelial or endocrine cells show clear cytoplasmic staining, sometimes with membrane-associated signal, at the reported High sites; isolated stromal colour or widespread staining of reported undetected cells needs control review (UniProt Q9HCY8: Cytoplasm; HPA tissue IHC: profile and cell-level results).
| Choice of positive tissue and cells | Use the named cells when assessing a control: squamous epithelial cells in esophagus or cervix, basal-layer cells in skin, or endocrine cells in colon are High (HPA tissue IHC). Whole-tissue averages can hide these restricted patterns (general IHC interpretation). |
| Compartment and topology | UniProt places S100A14 in the cytoplasm and reports no transmembrane segment (UniProt Q9HCY8). HPA also observes membrane-associated staining in some tissues (HPA tissue IHC: profile); assess that signal alongside the expected cell type. |
| IHC evidence and protocol scope | HPA rates tissue IHC reliability Enhanced and lists HPA027613 as IHC Enhanced (HPA tissue IHC; HPA antibodies). These ratings support pattern interpretation; neither supplies a target-specific retrieval condition or fixation-sensitivity claim. |
| IF/ICC Q&A: Should its compartments match IHC exactly? | No exact match is established: ICC-IF reports nuclear bodies, plasma membrane, cell junctions and cytosol, while HPA027613 is ICC Uncertain (HPA subcellular; HPA antibodies). Interpret IF/ICC with its own controls; this IHC section supplies no IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in esophageal or cervical squamous epithelium. | The assay may have failed; those cells are reported High in tissue IHC (HPA tissue IHC). | Check section integrity, antibody application and detection with an appropriate positive control (general IHC practice). Avoid inferring target-specific fixation sensitivity from this result. |
| Colon looks mostly unstained despite a few strongly coloured cells. | The expected High result belongs to endocrine cells, not all colon cells (HPA tissue IHC: Colon, Endocrine cells). | Inspect the positive cells at higher magnification and score their distribution by cell type (general IHC interpretation). |
| Staining is concentrated in stroma or extracellular deposits. | That pattern conflicts with the reported cellular distribution (UniProt Q9HCY8: Cytoplasm; HPA tissue IHC: profile). | Compare the expected epithelial cells and a negative control; review nonspecific staining before scoring the deposits as positive (general IHC practice). |
| Diffuse colour obscures cell boundaries throughout the section. | Background may arise from nonspecific antibody binding or endogenous detection activity (general IHC practice). | Review reagent-only controls and blocking or detection steps, then assess whether a cell-specific pattern remains (general IHC practice). |
| Reported undetected cells stain as strongly as the intended positive cells. | The cell-level pattern is discordant with HPA for adipocytes or bronchial respiratory epithelial cells (HPA tissue IHC: Not detected). | Verify cell identity and compare negative controls before interpreting the signal as S100A14 (general IHC practice). Do not generalize those negative calls to every cell in either tissue. |
| Nuclear-body signal in IF/ICC seems inconsistent with the IHC slide. | HPA reports nuclear bodies in ICC-IF, but lists HPA027613 ICC validation as Uncertain (HPA subcellular; HPA antibodies). | Evaluate IF/ICC with its own controls and validation evidence; do not use its nuclear-body signal alone to overturn an IHC pattern (general assay interpretation). |
Experiment evidence
IHC & IF images showing S100A14 staining
A06301-1 · IHCIHC analysis of S100A14 using anti-S100A14 antibody (A06301-1). S100A14 was detected in a paraffin-embedded section of human cervica squamou
A06301-1 · IHCIHC analysis of S100A14 using anti-S100A14 antibody (A06301-1). S100A14 was detected in a paraffin-embedded section of human lung squamous c
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000189334-S100A14/tissue/cerebral+cortex#imid_7356216
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000189334-S100A14/tissue/esophagus#imid_7356188
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000189334-S100A14/tissue/colon#imid_7353278
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000189334-S100A14/tissue/liver#imid_7353263
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027613; green = S100A14, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA027613; green = S100A14, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for S100A14 IHC & IF
🧪Run cervix first: squamous epithelial cells should stain (HPA: High in cervix squamous epithelial cells). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the cervix slide, cells without specific signal should remain at background rather than being assigned an unsupported cell identity.
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show S100A14 in A-431, HaCaT, MCF-7, with annotated localisation: Nuclear bodies (approved), Plasma membrane (approved), Cell Junctions (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a host- and clonality-matched rabbit IgG control (IHC caption: rabbit primary), and S100A14 knockout material as a biological negative. Quench endogenous peroxidase and check background in the cervix section before interpreting DAB staining (IHC caption: peroxidase secondary and DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06301-1 paraffin-section caption does not state the fixative (IHC caption: fixative unreported). The reported IHC staining used heat-mediated EDTA retrieval at pH 8.0; this documents a working retrieval condition without establishing that retrieval is required (IHC caption: EDTA pH 8.0). Whether frozen sections or IF are easier is unreported; for this chromogenic cervix assay, account for endogenous peroxidase background (IHC caption: peroxidase secondary and DAB detection).
HPA tissue IHC evidence for S100A14
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | 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 → |
| Bone marrow | Hematopoietic cells | 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 S100A14 IHC Tips
Troubleshoot S100A14 chromogenic IHC in paraffin sections using the catalog antibody’s tissue result and the supplied protein localisation evidence.
How should I adjust retrieval when S100A14 staining is weak or patchy?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A06301-1). Keep section thickness, heating time, cooling time and detection conditions constant while comparing retrieval runs; the selected image supplies no heating duration, so record the duration used locally (datasheet A06301-1; standard IHC practice). Include a positive squamous epithelial control, such as cervix, and inspect whether staining improves in the expected cells without widespread tissue damage (HPA: high in cervical squamous epithelial cells; standard IHC practice). If staining remains weak, compare a second validated retrieval condition on adjacent sections, changing one variable at a time and retaining the EDTA run as the reference (standard IHC practice).
Could fixation explain variable S100A14 staining between paraffin blocks?
The selected tissue image describes a paraffin-embedded section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A06301-1 caption). Record fixative, fixation duration, tissue thickness and processing history for each block before attributing differences to S100A14 biology (standard IHC practice). Compare blocks using the same EDTA pH 8.0 retrieval, 2 μg/ml primary antibody and overnight incubation at 4°C used in the selected image (datasheet A06301-1). Place a known positive tissue section in each run, then assess morphology and staining in the same cell type across blocks; this controls the comparison without assuming a S100A14-specific fixation effect (HPA: high in cervical squamous epithelial cells; standard IHC practice).
Should I reject membranous S100A14 staining as nonspecific?
No: cytoplasmic localisation is reported for S100A14, while tissue staining includes membranous expression in some settings (UniProt Q9HCY8 localisation; HPA: tissue IHC profile). Subcellular imaging also places S100A14 at the plasma membrane, cell junctions, cytosol and nuclear bodies, so annotate these compartments separately when reading chromogenic sections (HPA: subcellular summary). S100A14 has no transmembrane segment; a membranous pattern therefore does not establish that the protein spans the membrane (UniProt Q9HCY8 topology). Check whether the signal follows intact cell borders and appears in expected epithelial cells, then compare it with the no-primary control before scoring it as specific (HPA: high in cervical squamous epithelial cells; standard IHC practice).
How can I investigate unexpected staining if the antibody epitope is unspecified?
The supplied record lists a 104-amino-acid S100A14 chain, an EF-hand at residues 27–61 and no reported isoforms, but gives no epitope for this antibody (UniProt Q9HCY8 record; datasheet A06301-1 caption). Do not assign staining to a particular domain or protein variant without epitope mapping or independent evidence (standard IHC interpretation). The record reports no glycosylation sites or modified residues, which does not establish how fixation or retrieval affects antibody binding (UniProt Q9HCY8 record). Compare staining with an independently validated antibody against a documented distinct epitope, if available, while keeping the tissue region and scoring rules consistent; concordance supports an assignment but does not identify this antibody’s epitope (standard IHC practice).
How should I check S100A14 localisation by multiplex IF after seeing an IHC signal?
Use IF as a separate localisation check, pairing S100A14 with a marker for the expected squamous epithelial cells when examining cervical tissue (HPA: high in cervical squamous epithelial cells; standard IF practice). Choose fluorophores whose signals can be separated from each other and from the specimen’s autofluorescence, and inspect single-stain controls before interpreting overlap (standard IF practice). If the antibody epitope is cytosolic, test permeabilisation; for an accessible extracellular epitope, omit it, but do not assume either orientation because this antibody’s epitope is unspecified (HPA: cytosol and plasma membrane; datasheet A06301-1 caption; standard IF practice). Compare membrane, junctional and cytosolic signal with the chromogenic result while documenting the IF fixation and staining conditions independently (HPA: subcellular summary; standard IF practice).
What should I change when DAB appears outside the expected S100A14-positive cells?
First examine a no-primary control and the distribution of DAB before changing antibody concentration; diffuse staining in both sections points toward detection background (standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a peroxidase-conjugated secondary for 30 minutes at 37°C, and DAB development (datasheet A06301-1). A peroxidase block and careful washing are general chromogenic IHC steps; verify their performance when enzyme activity or residual reagent could contribute signal (standard IHC practice). Assess staining in expected squamous epithelial cells separately from low-expression lung macrophages or liver cholangiocytes, and avoid treating every brown cell as equally informative (HPA: high in cervical squamous epithelial cells; low in lung macrophages and liver cholangiocytes).
How can I score S100A14 staining across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measurement; cervical squamous epithelial cells are a supported positive population, and membrane and cytoplasmic signal may differ (HPA: high in cervical squamous epithelial cells; HPA: tissue IHC profile; UniProt Q9HCY8 localisation). For chromogenic IHC, report the percentage of positive target cells and an intensity-weighted H-score, using the same intensity categories and positivity threshold across sections (standard IHC practice). Normalise each result to the number of evaluable cells of that defined type, rather than all nuclei when cell composition varies (standard IHC practice). If scoring discrete positive cells, report density per mm² of evaluable tissue and exclude folds, necrosis and tissue-free areas using prespecified rules (standard IHC practice).
Which S100A14 patterns warrant validation before calling a section positive?
Give greatest weight to reproducible staining in the expected cell population: cervical squamous epithelial cells show high S100A14 staining in the supplied tissue profile (HPA: high in cervical squamous epithelial cells). Cytoplasmic or membranous staining can fit the reported distribution, whereas an isolated unexpected compartment needs confirmation with controls and an independent method (UniProt Q9HCY8 localisation; HPA: tissue IHC and subcellular summaries; standard IHC interpretation). Distrust signal restricted to section edges, folds or necrotic areas, especially if the no-primary control shows similar DAB deposition (standard IHC practice). Check the peroxidase block when brown signal follows endogenous enzyme activity, and compare the suspect section with a concurrently processed positive control before calling a biological difference (standard IHC practice).
Boster reagents
Best S100A14 / Protein S100-A14 IHC Antibodies
A06301-1 has real IHC data from paraffin sections of human cervical and lung squamous carcinomas (IHC image captions). Human is the only listed reactive species (catalog reactivity).
Real IHC data
Anti-S100A14 Antibody
Cat # A06301-1
A06301-1 is listed for human IHC (catalog applications and reactivity). Its images show staining in paraffin sections of human cervical and lung squamous carcinomas (IHC image captions).
Which to pick: Choose A06301-1 for human paraffin-section IHC: it is a rabbit polyclonal antibody listed for IHC, with a 2–5 µg/ml catalog range (catalog antibody details and dilution); its images use 2 µg/ml after EDTA pH 8.0 heat retrieval (IHC image captions). The captions identify paraffin sections but do not report the fixative (IHC image captions). No SKU in this payload is listed for IF/ICC or another species, so there is no supported pick for either (catalog applications and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9HCY8 (S10AE_HUMAN, Protein S100-A14).
- Human Protein Atlas. S100A14 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. S100A14 subcellular location (ICC-IF): Localized to the nuclear bodies, plasma membrane, cell junctions and cytosol..
- Human Protein Atlas. S100A14 antibody validation summary (1 antibodies).
- Co-expression of S100A14 and S100A16 correlates with a poor prognosis in human breast cancer and promotes cancer cell invasion. BMC cancer 2015 — PMC4348405.
- Overexpression of S100A14 contributes to malignant progression and predicts poor prognosis of lung adenocarcinoma. Thoracic cancer 2018 — PMC6026614.
- Calcium-binding protein S100A14 induces differentiation and suppresses metastasis in gastric cancer. Cell death & disease 2017 — PMC5550849.
- The role of S100A14 in epithelial ovarian tumors. Oncotarget 2014 — PMC4116497.
- PubMed PMID:11944983 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for S100A14 (Protein S100-A14). Expected localisation Cytoplasmic (UniProt); membranous in some tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06301-1). Validated IHC antibody: A06301-1 at 2-5 µg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human cervica squamous carcinoma tissue; fixative not specified (datasheet A06301-1); 2. Fixation: Image fixative and duration unreported (datasheet A06301-1); 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 A06301-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06301-1); 8. Primary antibody: Rabbit anti-S100A14, 2-5 µg/ml (datasheet A06301-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06301-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Whole-tissue scoring may obscure endocrine-cell staining (HPA tissue IHC); Isoform / epitope: One 1–104 chain, no isoforms; cytoplasmic epitope expected (UniProt); Regulation: Expression varies by cell type (HPA tissue IHC). Evidence: UniProt Q9HCY8, HPA tissue IHC · Enhanced, HPA ICC-IF · nuclear bodies, plasma membrane, cell junctions and cytosol., Datasheet A06301-1, PMC4348405, PMC6026614. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/S100A14/immunohistochemistry-ihc-guide.