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PCOLCE / Procollagen C-endopeptidase enhancer 1 · IHC design guide
Design Immunohistochemistry for PCOLCE
Plan PCOLCE staining in paraffin sections using breast glandular cells as a high-staining reference (HPA tissue IHC). The catalog antibody’s illustrated workflow uses 2 μg/ml primary antibody overnight at 4°C, followed by HRP/DAB detection (datasheet A06428-2).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PCOLCE 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 | Extracellular and cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Extracellular signal and cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06428-2) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Appendix+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 PCOLCE may stain beyond producing cells (UniProt) | |
| Regulation | Specific expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; signal peptide is cleaved (UniProt) |
Section 1
Recommended PCOLCE IHC & IF Protocols
The catalog antibody’s IHC protocol uses EDTA pH 8.0 retrieval (datasheet). Published PCOLCE IHC protocols describe glioma and renal carcinoma tissue (PMC11733849; PMC8740509).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A06428-2) |
| Fixation | Image fixative and duration unreported (datasheet A06428-2); 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 A06428-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06428-2) |
| Primary antibody | Rabbit anti-PCOLCE, 2-5 μg/ml (datasheet A06428-2) |
| Primary incubation | Overnight at 4 °C (datasheet A06428-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06428-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PCOLCE-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Extracellular positivity and cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 retrieval for the catalog antibody (datasheet); the glioma protocol used citrate pH 6.0 (PMC11733849).
Section 2
What Is the Expected PCOLCE Staining Pattern?
PCOLCE is secreted and has no transmembrane segment (UniProt Q15113 topology). In paraffin-section IHC, expect extracellular positivity and cytoplasmic staining in several tissues (HPA tissue IHC: Approved). Breast glandular cells provide a high-staining example; endometrial stromal cells and bronchial respiratory epithelial cells are medium-staining examples (HPA tissue IHC). Interpret cellular and extracellular signal together, since tissue RNA and protein locations can differ for this secreted protein (HPA reliability note).
| Extracellular staining accompanies glandular or stromal cytoplasmic staining. | This fits a secreted protein (UniProt Q15113 topology) and HPA's observed extracellular and cytoplasmic IHC profile (HPA tissue IHC). Judge each compartment against the tissue and cell type: breast glandular cells stain high, while endometrial stromal cells stain medium (HPA tissue IHC). |
| A crisp nuclear-only or membrane-rim pattern dominates the section. | Neither is the reported PCOLCE IHC pattern (HPA tissue IHC), and PCOLCE has no transmembrane segment (UniProt Q15113 topology). Treat the pattern as suspect; compare its distribution with the extracellular and cytoplasmic reference pattern before assigning specificity. |
| The strongest signal occurs in a cell population expected to be unstained. | For example, appendix glandular cells are not detected in HPA tissue IHC. Cross-reactivity or endogenous detection activity is plausible (general IHC practice). Check whether the signal persists when primary antibody is omitted, and compare it with a known-positive section. |
| Color spreads uniformly across cells, stroma, and empty-looking spaces. | Uniform haze cannot distinguish the extracellular positivity reported for PCOLCE from background (HPA tissue IHC; general IHC practice). Examine morphology, a primary-omission control, and a positive tissue processed in the same run before scoring extracellular staining. |
| Breast glandular cells show no detectable signal. | High staining is reported for these cells (HPA tissue IHC), so the negative result warrants a run check. Review tissue preservation, retrieval, antibody dilution, detection reagents, and the concurrent positive control as general IHC workflow checks; no PCOLCE-specific fixation sensitivity is established here. |
💡Expected PCOLCE appearanceCall the result positive when defined extracellular staining and cytoplasmic staining occur in the expected cells, with high signal in breast glandular cells or medium signal in endometrial stromal cells (HPA tissue IHC); isolated nuclear staining or uniform color haze is suspect (HPA tissue IHC; general IHC practice).
| Secretion and maturation | PCOLCE has a signal peptide at residues 1–25 and a mature chain at 26–449 (UniProt Q15113 processing). Its extracellular IHC signal can be spatially separate from stained cells (HPA tissue IHC). Do not infer the producing cell solely from extracellular deposits. |
| Tissue and cell baseline | HPA reports high breast glandular staining, medium endometrial stromal staining, and no detection in appendix glandular cells (HPA tissue IHC). These are reference observations for interpreting a run, not a claim that every section or cell must have identical intensity. |
| Antibody validation | The HPA tissue profile is Approved, and CAB017623 has Approved IHC status (HPA tissue IHC; HPA antibodies). Approval supports use of the reported pattern as a reference; it does not establish specificity for every specimen or detection system. |
| Epitope and retrieval limits | The supplied UniProt record identifies CUB and NTR domains and glycosylation sites at residues 29 and 431 (UniProt Q15113). It gives no antibody epitope or target-specific retrieval response, so choose retrieval conditions from the IHC-validated antibody's instructions and validate the resulting pattern (general IHC practice). |
| IF/ICC cross-check? | HPA reports approved Golgi and vesicle localization in ICC-IF, with U2OS images (HPA subcellular ICC-IF). This intracellular view can coexist with secretion and the extracellular tissue IHC profile (UniProt Q15113 topology; HPA tissue IHC). It does not define an IF protocol for this IHC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in the concurrent breast glandular positive section. | A run failure or insufficient assay signal is possible; breast glandular staining is high in the HPA reference (HPA tissue IHC). | Check antibody preparation and dilution, retrieval, detection reagents, and counterstain against the IHC antibody instructions (general IHC practice). Repeat with a concurrent positive section before interpreting test sections. |
| Only nuclei or sharply outlined cell membranes stain. | The location conflicts with extracellular and cytoplasmic PCOLCE tissue staining (HPA tissue IHC) and its lack of a transmembrane segment (UniProt Q15113 topology). | Review morphology and controls; reassess antibody specificity and detection background before scoring the signal as PCOLCE (general IHC practice). |
| Appendix glandular cells stain strongly. | Those cells are not detected in the HPA reference (HPA tissue IHC). Cross-reactivity or endogenous detection activity may explain the discrepancy (general IHC practice). | Run a primary-omission control and inspect the detection system's endogenous-activity blocking step (general IHC practice). Compare the result with breast glandular cells in the same run (HPA tissue IHC). |
| Extracellular staining is diffuse and hard to separate from haze. | Extracellular positivity is expected (HPA tissue IHC), but widespread nonspecific color can obscure its boundaries (general IHC practice). | Compare signal with the primary-omission control, tissue morphology, and expected stained cells. Review blocking, washes, and detection development as general IHC checks; do not score uniform haze alone. |
| A positive tissue stains, but the test tissue is weak or negative. | PCOLCE tissue levels vary: breast glandular cells are high, whereas lung alveolar cells are low (HPA tissue IHC). A working positive control does not establish expression in the test cell population. | Identify the exact tissue and cell population, then compare its HPA reference level. Score the observed compartment and intensity without extending the positive control's expected intensity to another tissue. |
| An ICC-IF image shows Golgi or vesicles while tissue IHC shows extracellular signal. | Both locations are reported: approved Golgi and vesicle ICC-IF localization (HPA subcellular ICC-IF) and extracellular tissue positivity (HPA tissue IHC); PCOLCE is secreted (UniProt Q15113 topology). | Interpret each application against its own HPA reference. For paraffin-section IHC, assess extracellular and cytoplasmic staining in the relevant tissue cells (HPA tissue IHC); use the separate IF/ICC guide for that application's setup. |
Experiment evidence
IHC & IF images showing PCOLCE staining
A06428-2 · IHCIHC analysis of PCOLCE using anti-PCOLCE antibody (A06428-2). PCOLCE was detected in a paraffin-embedded section of human tonsil tissue. Hea
A06428-2 · IHCIHC analysis of PCOLCE using anti-PCOLCE antibody (A06428-2). PCOLCE was detected in a paraffin-embedded section of human thyroid papillary
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106333-PCOLCE/tissue/cerebral+cortex#imid_4490556
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106333-PCOLCE/tissue/colon#imid_4490498
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106333-PCOLCE/tissue/liver#imid_4490499
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106333-PCOLCE/tissue/kidney#imid_4490500
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA042927; green = PCOLCE, blue = nucleus, red = microtubules.
HPA · ICC-IF U2OSHuman Protein Atlas immunofluorescence, U2OS, antibody HPA042927; green = PCOLCE, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PCOLCE IHC & IF
🧪Run breast first: glandular cells should stain (HPA: breast glandular cells, High). Run appendix glandular cells as the negative tissue (HPA: appendix glandular cells, Not detected); on the breast slide, internal negative cells should show only background signal, although HPA does not identify a specific negative breast cell type.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PCOLCE in U2OS, with annotated localisation: Golgi apparatus (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a host- and clonality-matched rabbit IgG isotype control where the primary’s clonality is known (caption: rabbit anti-PCOLCE A06428-2); and a PCOLCE knockout sample or immunizing-peptide block, if available. Quench endogenous peroxidase and check background in breast tissue before interpreting DAB staining (caption: breast control uses peroxidase/DAB detection).
⚠️Feasibility: Paraffin-section IHC is demonstrated with heat retrieval in EDTA at pH 8.0 (caption: A06428-2), but a target-specific fixation window or fixation effect is unreported, and the selected caption does not state the fixative. The supplied evidence does not establish that frozen sections or IF are easier; Golgi and vesicle staining has been observed by ICC-IF in U2OS cells (HPA: subcellular localization). In breast, assess glandular staining separately from extracellular or luminal signal because PCOLCE is secreted (UniProt Q15113: secreted).
HPA tissue IHC evidence for PCOLCE
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Secreted protein, tissue location of RNA and protein is expected to differ.). 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PCOLCE IHC Tips
Troubleshoot PCOLCE staining by assessing retrieval, compartment, and detection alongside matched tissue controls (UniProt Q15113; HPA tissue IHC).
What retrieval should I use when PCOLCE staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin-section IHC (datasheet A06428-2). The selected tonsil image used this retrieval before an overnight primary incubation at 4°C (datasheet A06428-2). If staining is weak, check that heating and cooling were consistent across slides, then compare retrieval times on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Include a no-primary control and inspect tissue structure, because excessive heating can make weak extracellular staining difficult to distinguish from damaged tissue (standard IHC practice). Score the same compartments across conditions: PCOLCE can appear extracellularly and in cytoplasm (HPA tissue IHC).
Could fixation explain variable PCOLCE staining between paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A06428-2). Record each block’s fixative and fixation interval, and compare adjacent sections using the same pH 8.0 EDTA retrieval and detection conditions (datasheet A06428-2; standard IHC practice). Check tissue preservation and staining in a control section from each block before attributing a negative result to PCOLCE biology (standard IHC practice). If blocks differ, optimize retrieval on representative sections and report the handling difference with the scores (standard IHC practice). Neither the reported tissue pattern nor PCOLCE topology establishes a target-specific fixation effect (HPA tissue IHC; UniProt Q15113).
Where should PCOLCE signal appear, and how should I assess cytoplasmic staining?
Assess extracellular deposits separately from cytoplasmic staining, because both occur in tissue IHC (HPA tissue IHC). PCOLCE is secreted and has no transmembrane segment, so a sharply continuous membrane-only pattern needs scrutiny before it is assigned to this target (UniProt Q15113 topology). Its signal peptide occupies residues 1–25, while the annotated mature chain spans 26–449 (UniProt Q15113 processing). Intracellular signal can be compatible with production or trafficking, and Golgi and vesicle localization has been reported by IF (HPA subcellular). Compare cell-associated and extracellular compartments on the same section, using morphology and a no-primary control to identify misplaced chromogen (standard IHC practice).
How could epitope choice change the PCOLCE IHC pattern?
Check the antibody’s stated immunogen or epitope before interpreting a negative region; the selected tissue-IHC caption does not identify one (datasheet A06428-2). UniProt lists 0 isoforms and annotates CUB domains at 37–149 and 159–273, plus an NTR domain at 318–437 (UniProt Q15113). PCOLCE also has annotated glycosylation sites at 29 and 431, but their effect on this antibody’s staining is unknown (UniProt Q15113; datasheet A06428-2). If two antibodies disagree, compare their disclosed epitopes, retrieval conditions, and extracellular versus cellular scores on adjacent sections (standard IHC practice). Do not interpret discordant staining as isoform-specific without supporting epitope evidence (UniProt Q15113; standard IHC practice).
How can IF help assess a disputed PCOLCE IHC localization?
Use IF as a separate localization check, rather than transferring the paraffin-section IHC conditions to it (datasheet A06428-2; standard IF practice). Multiplex PCOLCE with a marker for the cell population being assessed, such as a glandular epithelial marker when examining glandular signal (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and test an unstained section for autofluorescence; a far-red channel may improve contrast when shorter wavelengths are bright (standard IF practice). For intracellular Golgi or vesicle epitopes, assess permeabilisation, whereas extracellular signal should also be examined without permeabilisation (HPA subcellular; UniProt Q15113 topology; standard IF practice). No IF performance claim for the catalog antibody follows from its IHC caption (datasheet A06428-2).
How do I reduce diffuse brown staining without losing extracellular PCOLCE?
Start by checking a no-primary slide and the distribution of brown precipitate, since the selected IHC image used peroxidase detection with DAB (datasheet A06428-2; standard IHC practice). Block endogenous peroxidase and nonspecific binding, then keep wash conditions identical across the test and control sections (standard IHC practice). The selected tonsil method used 10% goat serum and 2 μg/ml primary antibody overnight at 4°C (datasheet A06428-2). If background persists, titrate the primary around that documented concentration while monitoring tissue morphology and extracellular staining (datasheet A06428-2; standard IHC practice). Do not discard all diffuse signal automatically, because extracellular PCOLCE positivity is reported in tissue (HPA tissue IHC).
How should I quantify PCOLCE across specimens with different stromal content? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions before scoring, and record cellular and extracellular signal separately because both are reported for PCOLCE (HPA tissue IHC; standard IHC practice). For identifiable cells, report the percentage positive and an H-score from staining intensity and positive-cell proportions; keep thresholds fixed across slides (standard IHC practice). For extracellular signal, measure positive area or optical density per mm² of viable tissue within a consistently defined compartment (standard IHC practice). Normalize cell scores to the number of eligible cells and extracellular scores to eligible tissue area, while recording stromal fraction and excluded damaged regions (standard IHC practice). Compare only sections processed and imaged under matched conditions, with controls used to set the background threshold (standard IHC practice).
When is a strong PCOLCE stain likely to be artefactual?
Treat extracellular and cytoplasmic staining as plausible patterns, but investigate a membrane-only pattern because PCOLCE is secreted and lacks a transmembrane segment (HPA tissue IHC; UniProt Q15113 topology). Check whether positive cells match the annotated population: breast glandular cells are high, whereas appendix glandular cells are not detected in the cited survey (HPA tissue IHC). Inspect edges, folds, and necrotic areas for localized precipitate, then compare a no-primary control to identify detection background (standard IHC practice). Endogenous peroxidase can mimic DAB positivity, so confirm that peroxidase blocking removes control staining before interpreting a strong signal (standard IHC practice). A negative cell compartment does not exclude extracellular PCOLCE deposition (UniProt Q15113 subcellular; HPA tissue IHC).
Boster reagents
Best PCOLCE / Procollagen C-endopeptidase enhancer 1 IHC Antibodies
The catalog contains one human-reactive anti-PCOLCE antibody with paraffin-section IHC images from human tonsil and thyroid papillary carcinoma; no IF image is supplied (catalog: A06428-2 image captions).
Real IHC data
Anti-PCOLCE Antibody ®
Cat # A06428-2
A06428-2 is listed for human IHC (catalog: A06428-2 applications and reactivity). Its image captions show staining in paraffin sections of human tonsil and thyroid papillary carcinoma (catalog: A06428-2 IHC image captions).
Which to pick: Choose A06428-2 for human paraffin-section IHC; its caption documents EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (catalog: A06428-2 tonsil IHC caption). Its application list provides no IF/ICC validation, and its reactivity is limited to human, so the payload supports neither an IF/ICC nor a cross-species recommendation (catalog: A06428-2 applications and reactivity). The caption identifies paraffin sections but does not report the fixative (catalog: A06428-2 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15113 (PCOC1_HUMAN, Procollagen C-endopeptidase enhancer 1).
- Human Protein Atlas. PCOLCE tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PCOLCE subcellular location (ICC-IF): Localized to the Golgi apparatus and vesicles..
- Human Protein Atlas. PCOLCE antibody validation summary (2 antibodies).
- Procollagen C-protease enhancer protein promotes glioma growth by activating ERK signaling. Asia-Pacific journal of clinical oncology 2025 — PMC11733849.
- Key genes associated with prognosis and metastasis of clear cell renal cell carcinoma. PeerJ 2022 — PMC8740509.
- Hind limb unloading of mice modulates gene expression at the protein and mRNA level in mesenchymal bone cells. BMC musculoskeletal disorders 2010 — PMC2906435.
- Predictive value of procollagen c-protease enhancer protein on the prognosis of glioma patients. Heliyon 2024 — PMC10963382.
- PubMed PMID:7523404 — UniProt-cited evidence.
- PubMed PMID:9799793 — UniProt-cited evidence.
- PubMed PMID:9933570 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PCOLCE (Procollagen C-endopeptidase enhancer 1). Expected localisation Extracellular and cytoplasmic staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06428-2). Validated IHC antibody: A06428-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Appendix. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A06428-2); 2. Fixation: Image fixative and duration unreported (datasheet A06428-2); 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 A06428-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06428-2); 8. Primary antibody: Rabbit anti-PCOLCE, 2-5 μg/ml (datasheet A06428-2); 9. Primary incubation: Overnight at 4 °C (datasheet A06428-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Secreted PCOLCE may stain beyond producing cells (UniProt); Isoform / epitope: No annotated isoforms; signal peptide is cleaved (UniProt); Regulation: Specific expression regulation is not annotated (UniProt). Evidence: UniProt Q15113, HPA tissue IHC · Approved, HPA ICC-IF · Golgi apparatus and vesicles., Datasheet A06428-2, PMC11733849, PMC8740509. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PCOLCE/immunohistochemistry-ihc-guide.