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SPINK7 / Serine protease inhibitor Kazal-type 7 · IHC design guide
Design Immunohistochemistry for SPINK7
Plan chromogenic IHC for SPINK7 using cytoplasmic staining in epithelial cells as the tissue reference (HPA tissue IHC). Compare staining across sections fixed consistently, while allowing for differences between protein and RNA location because SPINK7 is secreted (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SPINK7 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 several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in squamous and glandular cells and keratinocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Secreted protein may stain away from its site of synthesis (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 20–85 (UniProt) |
Section 1
Recommended SPINK7 IHC & IF Protocols
The catalog antibody protocol is followed by one published SPINK7 IHC protocol using skin wound tissue (PMC11949503).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A11563); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Chicken anti-SPINK7, 5 μg/mL (datasheet A11563) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SPINK7-positive staining in squamous epithelial cells of cervix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval setting), then optimize for the tissue.
Section 2
What Is the Expected SPINK7 Staining Pattern?
SPINK7 is secreted and has no transmembrane segment (UniProt P58062 topology). In tissue IHC, expect mainly cytoplasmic staining in epithelial cells, including cervical and esophageal squamous cells and intestinal glandular cells (HPA: tissue IHC). HPA rates its tissue staining Enhanced, while noting medium agreement with RNA expression and pending external verification (HPA: reliability description).
| Moderate cytoplasmic staining in cervical or esophageal squamous epithelial cells. | This matches the reported positive cell types and intensity (HPA: Cervix and Esophagus, Medium). A secreted protein can also appear outside its producing cells, so assess any extracellular signal against the epithelial pattern and controls (UniProt P58062: Secreted; general IHC practice). |
| Strong nuclear staining dominates while the expected epithelial cytoplasm is unstained. | Treat this as a suspect compartment pattern: HPA describes cytoplasmic expression, and UniProt describes a secreted protein (HPA: tissue profile; UniProt P58062: Secreted). Review controls and detection conditions before assigning the nuclear signal to SPINK7 (general IHC practice). |
| Staining is prominent in an unexpected cell type but absent from nearby expected epithelial cells. | Consider cross-reactivity or endogenous detection activity (general IHC practice). Check the specific tissue first: HPA reports Low staining in lung macrophages, so macrophage signal there is not categorically unexpected (HPA: Lung, macrophages, Low). |
| Color is spread across the section without a clear cell-associated pattern. | Diffuse background limits interpretation of the reported cytoplasmic epithelial signal (HPA: tissue profile; general IHC practice). Compare a no-primary control and inspect wash and detection conditions before scoring the section (general IHC practice). |
| No staining appears in cervical or esophageal squamous epithelium. | These are reported Medium-staining reference tissues, so a blank result warrants a run check (HPA: Cervix and Esophagus, Medium). Confirm tissue preservation, antibody application, retrieval conditions and detection controls; no SPINK7-specific retrieval condition is supplied (general IHC practice). |
💡Expected SPINK7 appearanceCall a positive result when epithelial cells show predominantly moderate cytoplasmic staining in an HPA-positive tissue (HPA: tissue profile; Cervix and Esophagus, Medium); isolated strong nuclear color or uniform section-wide color is suspect (HPA: tissue profile; general IHC practice).
| Tissue and cell selection | Cervical, esophageal and oral squamous epithelial cells and intestinal glandular cells are reported at Medium intensity (HPA: tissue IHC). Adipocytes in adipose tissue and respiratory epithelial cells in bronchus are Not detected; use those observations as context, not proof that every cell in those tissues is negative (HPA: Adipose tissue and Bronchus). |
| Secretion and processing | SPINK7 has a signal peptide at residues 1–19 and a mature chain at 20–85 (UniProt P58062: processing). Its secreted status makes extracellular distribution plausible, but the supplied sources do not specify an extracellular IHC pattern (UniProt P58062: Secreted; HPA: cytoplasmic tissue profile). |
| Antibody evidence | The listed rabbit antibody HPA036294 has Enhanced IHC validation (HPA: antibody validation). HPA also reports medium consistency between staining and RNA data and says external verification is pending; evaluate any unexpected pattern with controls (HPA: reliability description; general IHC practice). |
| IF/ICC: what localisation is established? | HPA lists SPINK7 as secreted but supplies no main subcellular location or ICC-IF image-bearing cell line (HPA: subcellular record). Its tissue IHC cytoplasmic pattern can guide interpretation, but these data do not establish a cell-line IF pattern (HPA: tissue profile and subcellular record). |
| Retrieval and detection | Antigen retrieval and endogenous detection activity are general paraffin-IHC considerations (general IHC practice). Neither source supplies a SPINK7-specific retrieval setting or fixation-sensitivity result; use control sections when assessing a changed signal (HPA: supplied IHC record; UniProt P58062: supplied record; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive epithelium is blank. | A failed staining step or an unsuitable run condition is possible (general IHC practice); HPA reports Medium staining in cervical and esophageal squamous cells (HPA: tissue IHC). | Run a known-positive section alongside the sample; check antibody application, retrieval and detection steps, then repeat with controlled adjustments (general IHC practice). |
| The entire section has diffuse color. | Background from detection chemistry, inadequate washing or nonspecific antibody binding can obscure cell-associated staining (general IHC practice). | Compare a no-primary control, review blocking and wash steps, and reassess staining only where cell boundaries and cytoplasm are discernible (general IHC practice; HPA: cytoplasmic tissue profile). |
| Nuclei stain more strongly than epithelial cytoplasm. | This conflicts with HPA's cytoplasmic tissue profile and is suspect for nonspecific staining or detection artifact (HPA: tissue profile; general IHC practice). | Check no-primary and known-positive controls, then review antibody and detection conditions before scoring nuclear color as SPINK7 (general IHC practice). |
| Unexpected cells stain while expected epithelium does not. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); low lung macrophage staining is reported and needs tissue-specific interpretation (HPA: Lung, macrophages, Low). | Identify the stained cell type, compare the matching HPA tissue entry, and use appropriate detection controls before calling the result specific (HPA: tissue IHC; general IHC practice). |
| Extracellular color accompanies epithelial staining. | Secretion makes extracellular SPINK7 plausible, but HPA describes the observed tissue profile as cytoplasmic (UniProt P58062: Secreted; HPA: tissue profile). | Score the epithelial cytoplasmic pattern separately; check whether extracellular color persists in the no-primary control before interpreting it (HPA: tissue profile; general IHC practice). |
| An IF/ICC image lacks the expected IHC pattern. | HPA supplies no ICC-IF image-bearing cell line or defined main subcellular location for SPINK7 (HPA: subcellular record). | Avoid treating the tissue IHC pattern as an established IF/ICC benchmark; interpret IF/ICC with its own controls and evidence (HPA: tissue profile and subcellular record; general IF practice). |
Experiment evidence
IHC & IF images showing SPINK7 staining
A11563 · IHCImmunohistochemistry of ECRG2 in rat lung tissue with ECRG2 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000145879-SPINK7/tissue/cerebral+cortex#imid_8930666
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000145879-SPINK7/tissue/colon#imid_8930592
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000145879-SPINK7/tissue/liver#imid_8930577
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000145879-SPINK7/tissue/kidney#imid_8930578
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SPINK7 IHC & IF
🧪Run esophagus first: squamous epithelial cells should stain (HPA: Esophagus, squamous epithelial cells, Medium). Use bronchus respiratory epithelial cells as the negative tissue (HPA: Bronchus, respiratory epithelial cells, Not detected); on the esophagus slide, cells outside the stained squamous epithelial compartment should show background-level chromogen, but confirm their negative status empirically.
Positive control tissue: Cervix (Squamous epithelial 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 SPINK7; derive a cell-line control from the positive tissue's cell type (Squamous epithelial cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) slide and an isotype control matched to the primary antibody’s host species, clonality and concentration; use knockout tissue or peptide blocking, if available, to test target specificity (standard IHC practice). Quench endogenous peroxidase for chromogenic detection and check whether the detection system requires endogenous-biotin blocking; inspect the esophagus control slides for background signal (standard IHC practice).
⚠️Feasibility: No supplied matched source reports a SPINK7-specific fixation window or retrieval effect, so optimize antigen retrieval empirically; the A11563 rat-lung IHC caption does not state a fixative (caption: fixative unreported). The evidence does not establish that frozen sections or IF are easier, and HPA lists no ICC-IF image cell lines (HPA subcellular: no cell lines with ICC-IF images). In esophagus, check tissue folds and section edges for nonspecific chromogen before scoring squamous epithelial staining (standard IHC practice).
HPA tissue IHC evidence for SPINK7
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. At least one protein variant secreted, tissue location of RNA and protein might differ and correlation is complex. 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 |
|---|---|---|---|---|
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SPINK7 IHC Tips
Troubleshoot SPINK7 staining in chromogenic paraffin section IHC using the supplied tissue evidence and its secreted protein biology.
How should I optimize retrieval when SPINK7 staining is weak?
Begin with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval setting). Run a known positive section beside the test section, then compare signal and tissue preservation before changing conditions (standard IHC practice; HPA: medium staining in esophageal squamous epithelial cells). If staining remains weak, test a modestly longer retrieval time on adjacent sections while keeping the buffer and detection method constant (standard IHC practice). Record whether improvement occurs in the expected cells rather than uniformly across the section; SPINK7 is secreted, so diffuse signal alone is insufficient evidence of specificity (UniProt P58062: secreted; HPA: tissue IHC profile).
Could fixation explain variable SPINK7 staining between sections?
Target-specific sensitivity to fixation is unknown from the supplied evidence; evaluate fixation as an experimental variable without assuming a SPINK7-specific effect. The catalog image reports rat lung IHC at 5 µg/mL but does not state its fixative (catalog antibody A11563 caption). Compare sections with documented processing histories using the same citrate pH 6.0 retrieval and detection run (page retrieval setting; standard IHC practice). Include a positive control section and assess morphology alongside staining, because poor preservation or uneven processing can complicate chromogenic IHC interpretation (HPA: medium staining in esophageal squamous epithelial cells; standard IHC practice).
Where should convincing SPINK7 signal appear in tissue?
Assess staining in the cell types supported by tissue evidence: HPA reports medium signal in esophageal and cervical squamous epithelial cells and in several glandular populations (HPA: tissue IHC). Its tissue profile describes cytoplasmic expression, while UniProt identifies SPINK7 as secreted with no transmembrane segment (HPA: cytoplasmic expression; UniProt P58062: topology). Cytoplasmic staining can therefore be plausible, but do not require a sharply bounded membrane pattern or interpret every extracellular deposit as specific (HPA: tissue IHC profile; UniProt P58062: secreted). Compare the candidate signal with adjacent morphology and a no-primary control, especially where secreted material may obscure cell-of-origin assignment (standard IHC practice).
How do processing and epitope location affect SPINK7 staining?
SPINK7 is an 85-amino-acid precursor with a signal peptide at residues 1–19 and a mature chain at 20–85 (UniProt P58062: processing). The Kazal-like domain spans residues 26–85, and the supplied record lists no alternative isoforms or glycosylation sites (UniProt P58062: domains, isoforms, glycosylation). Check the antibody's documented immunogen or epitope before attributing absent staining to processing, since its epitope is not supplied here (standard IHC practice). When comparing antibodies, test them on adjacent positive sections under matched retrieval and detection conditions; agreement in expected cells is more informative than intensity alone (HPA: tissue IHC; standard IHC practice).
How would I check a SPINK7 IHC pattern by multiplex IF?
For a separate IF experiment, pair SPINK7 with a validated marker of the expected epithelial or glandular cell population and inspect cellular overlap (HPA: positive cell types; standard IF practice). Choose spectrally separated fluorophores after checking unstained tissue autofluorescence, and include single-stain controls before interpreting a merged image (standard IF practice). If the antibody recognizes an intracellular epitope, optimize mild permeabilisation; if it recognizes accessible extracellular secreted protein, compare with an unpermeabilised condition (UniProt P58062: secreted, no transmembrane segment; standard IF practice). Because no SPINK7 ICC/IF images or epitope location are supplied, treat the IF pattern as an independently validated result, not an automatic extension of the IHC image (HPA: subcellular record; catalog antibody A11563 caption).
How can I separate SPINK7 signal from chromogenic background?
Run a no-primary control and inspect diffuse deposits, section edges, folds and blood-rich regions before assigning brown signal to SPINK7 (standard IHC practice). Use a peroxidase block for HRP detection, then compare DAB development times across matched sections so an overdeveloped slide does not define the threshold (standard IHC practice). If background persists, optimize blocking, antibody concentration and wash conditions one variable at a time; the reported 5 µg/mL applies to the catalog rat lung image, not every tissue (catalog antibody A11563 caption; standard IHC practice). Retain a positive tissue control while adjusting conditions, since expected signal can be modest in some sites (HPA: low staining in lung macrophages).
What scoring method best captures variable SPINK7 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; HPA reports cytoplasmic expression across several tissues and medium staining in selected epithelial and glandular cells (HPA: tissue IHC profile). For cell-associated signal, record percentage positive and intensity, or calculate an H-score from prespecified intensity bins (standard IHC practice). Normalize positive-cell counts to the number of evaluable cells in the same compartment; for spatial deposits, report stained area or density per mm² of evaluable tissue (standard IHC practice). Apply one threshold across batches using matched controls, and report the denominator and excluded damaged areas so differences in tissue composition do not masquerade as SPINK7 changes (standard IHC practice).
What would make an apparent SPINK7-positive section unconvincing?
A convincing pattern should align with tissue morphology and supported cell types, such as esophageal squamous epithelial cells, rather than appearing only in damaged margins (HPA: tissue IHC; standard IHC practice). Interpret strong signal in an unexpected compartment cautiously because SPINK7 is secreted and lacks a transmembrane segment, while the reported tissue pattern is cytoplasmic (UniProt P58062: topology; HPA: tissue IHC profile). Check edge effects, necrotic regions and endogenous peroxidase with morphology and controls before treating DAB deposits as antigen signal (standard IHC practice). HPA rates its tissue evidence Enhanced but notes medium RNA–protein consistency and pending external verification, so corroborate disputed patterns with an independent control (HPA: reliability description; standard IHC practice).
Boster reagents
Best SPINK7 / Serine protease inhibitor Kazal-type 7 IHC Antibodies
A11563 has real IHC data from rat lung tissue at 5 μg/mL (A11563 image caption). Human, mouse and rat are listed as reactive; no IF image is provided (catalog reactivity/images).
Real IHC data
Anti-ECRG2 SPINK7 Antibody
Cat # A11563
A11563 is the sole card and shows IHC staining of rat lung tissue at 5 μg/mL (A11563 image caption). It lists IHC-P and human, mouse and rat reactivity, while the datasheet reports IHC validation in rat samples (catalog applications/reactivity; datasheet validation).
Which to pick: Choose A11563 for paraffin-section tissue IHC: IHC-P is listed, and its own image shows rat lung tissue at 5 μg/mL (catalog applications; A11563 image caption). No SKU is supported for IF/ICC; A11563 has no IF/ICC application or image, and although human and mouse reactivity is listed, IHC validation is reported only in rat samples (catalog applications/images/reactivity; datasheet validation). Clonality and the fixative used for the pictured tissue are unreported (catalog clone; A11563 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P58062 (ISK7_HUMAN, Serine protease inhibitor Kazal-type 7).
- Human Protein Atlas. SPINK7 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SPINK7 subcellular location (ICC-IF): Secreted.
- Human Protein Atlas. SPINK7 antibody validation summary (1 antibodies).
- The antiprotease Spink7 promotes inflammation resolution by modulating multiple proteases activities during wound healing. Clinical and translational medicine 2025 — PMC11949503.
- Development and Validation of a Centrosome Amplification-Related Prognostic Model in Pancreatic Cancer: Multi-Omics Guided Risk Stratification and Tumor Microenvironment. Cancers 2025 — PMC12468691.
- PubMed PMID:12975309 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:22528291 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SPINK7 (Serine protease inhibitor Kazal-type 7). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A11563 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Squamous epithelial cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A11563); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Chicken anti-SPINK7, 5 μg/mL (datasheet A11563); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Secreted protein may stain away from its site of synthesis (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 20–85 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P58062, HPA tissue IHC · Enhanced, HPA ICC-IF · Secreted, Datasheet A11563, PMC11949503. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SPINK7/immunohistochemistry-ihc-guide.