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APC / Adenomatous polyposis coli protein · IHC design guide
Design Immunohistochemistry for APC
Plan APC staining in paraffin sections using the catalog antibody’s documented IHC workflow. Compare cytoplasmic and membranous staining in the cell types reported by HPA, and account for tissue-specific differences when interpreting signal (datasheet M00008; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
APC 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular and epithelial cells show cytoplasmic and membranous staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00008) | |
| 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 paraffin-section fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Cortex neuropil stains highly; caudate glia are undetected (HPA tissue IHC) | |
| Regulation | HGF shifts APC toward cell protrusions (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended APC IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: M00008) is accompanied by published protocols for liver and colorectal cancer sections (PMC2757596; PMC5603343).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet M00008) |
| Fixation | Image fixative and duration unreported (datasheet M00008); 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 M00008); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00008) |
| Primary antibody | Rabbit monoclonal (clone FAD-1) anti-APC, 1:50 (datasheet M00008) |
| Primary incubation | Overnight at 4 °C (datasheet M00008) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00008) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | APC-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: M00008); the colorectal article used Tris-EDTA pH 9.0 (PMC5603343).
Section 2
What Is the Expected APC Staining Pattern?
APC is expected mainly in the cytoplasm and along cell membranes in tissue IHC (HPA tissue IHC: general cytoplasmic and membranous expression). Look within identified cells, such as glandular cells in appendix or duodenum and cholangiocytes in liver (HPA: High in these cells). APC also associates with adherens junctions and the cytoskeleton, and has no transmembrane segment (UniProt P25054: subcellular location; topology). HPA rates the tissue IHC pattern Supported, with medium consistency between staining and RNA data (HPA tissue IHC: reliability).
| Cytoplasmic staining with a membranous component in glandular cells of appendix or duodenum (HPA: High in these cells). | This fits the reported tissue pattern (HPA tissue IHC: general cytoplasmic and membranous expression). Score the identified glandular cells and their compartments; a uniformly stained section is harder to interpret. |
| Cytoplasmic or membranous staining in cholangiocytes, with staining assessed separately from surrounding liver cells (HPA: High in cholangiocytes). | The reported High result names cholangiocytes specifically (HPA: High in liver cholangiocytes). Do not extend that cell-level result to every cell in the liver section. |
| Predominantly nuclear staining, without the expected cytoplasmic or membranous pattern (HPA tissue IHC: general cytoplasmic and membranous expression). | Treat this as discordant with the reported IHC pattern and check assay specificity or background (HPA tissue IHC: general cytoplasmic and membranous expression; general IHC practice). |
| Strong staining in adipocytes or cardiomyocytes (HPA: Not detected in adipocytes and cardiomyocytes). | This conflicts with those cell-level HPA observations and warrants a cross-reactivity or endogenous detection check (HPA: Not detected in those cells; general IHC practice). It does not prove every cell in either tissue is APC-negative. |
| Widespread diffuse chromogen, or no staining in appendix glandular cells used as a positive tissue (HPA: High in appendix glandular cells). | Diffuse deposit obscures cell and compartment scoring; an absent positive-tissue signal leaves the run uninterpretable (general IHC practice). Check detection background or assay performance before calling APC absent. |
💡Expected APC appearanceA convincing positive shows clear cytoplasmic staining, often with a membranous component, in identified High-staining cells such as appendix glandular cells; diffuse section-wide deposit or nuclear-only signal is suspect (HPA: High in appendix glandular cells; HPA tissue IHC: general cytoplasmic and membranous expression; general IHC practice).
| Compartment and topology (UniProt P25054: subcellular location; topology). | APC is associated with junctions, cytoskeleton, cytoplasm and cell membrane, but has no transmembrane segment (UniProt P25054: subcellular location; topology). Membranous IHC therefore need not imply a membrane-spanning protein. |
| Cell-level tissue variation (HPA tissue IHC: positive and negative cells). | HPA reports High staining in kidney proximal tubule cell bodies but Not detected in adipocytes (HPA: kidney; adipose tissue). Choose and score named cell populations; a tissue-wide positive or negative label would overstate these observations. |
| Isoforms and epitope scope (UniProt P25054: isoforms; tissue specificity). | UniProt lists isoforms 1A, 2 and 1B; 1A is especially expressed in brain, while 1B predominates in most other tissues (UniProt P25054: isoforms; tissue specificity). Without an antibody epitope in the payload, isoform-specific staining cannot be predicted. |
| IHC validation scope (HPA antibodies: HPA013349 IHC Supported; HPA tissue IHC: Supported). | The supplied IHC antibody status is Supported, and tissue staining has medium consistency with RNA data (HPA antibodies: HPA013349; HPA tissue IHC: reliability). These labels support pattern review, not a claim that every positive deposit is specific. |
| What does IF/ICC add? (HPA subcellular ICC-IF: plasma membrane supported; Golgi approved). | HPA IF/ICC places APC mainly at the plasma membrane, with additional Golgi localization (HPA subcellular ICC-IF). This is a separate cell-imaging observation; evaluate paraffin IHC against the supplied tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in appendix glandular cells expected to stain High (HPA: appendix glandular cells High). | The positive-tissue control has failed, or the assay may be insensitive (general IHC practice); HPA staining alone does not identify which assay step failed. | Verify tissue identity and cell selection, then review the IHC-validated antibody's supplied protocol and detection controls (general IHC practice). Do not score other sections negative until the positive control works. |
| Nuclear-only staining despite the reported cytoplasmic and membranous IHC profile (HPA tissue IHC: profile). | The observed compartment disagrees with the reference pattern; nonspecific binding or detection background is possible (HPA tissue IHC: profile; general IHC practice). | Compare a matched negative detection control and the expected positive-cell pattern before accepting nuclear staining (general IHC practice). Record the discordance rather than assigning a new APC location. |
| Strong adipocyte or cardiomyocyte staining (HPA: Not detected in adipocytes and cardiomyocytes). | Cross-reactivity or endogenous detection activity is possible (general IHC practice); the HPA result applies to the named cells, not every cell in those tissues. | Check cell identification and a matched negative detection control, then compare with a named High-staining cell population (HPA: positive and negative cells; general IHC practice). |
| Diffuse chromogen across cells and extracellular areas, obscuring the reported compartment pattern (HPA tissue IHC: profile). | Detection background or inadequate blocking may obscure localization (general IHC practice). The diffuse deposit cannot establish cell-specific APC expression. | Review blocking, washes, chromogen development and a negative detection control (general IHC practice). Rescore only when cell boundaries and cytoplasmic or membranous staining can be distinguished. |
| A kidney section appears negative despite the reported High result in proximal tubule cell bodies (HPA: kidney proximal tubules High). | The scored field may lack identifiable proximal tubules, or the run may have lost sensitivity (HPA: kidney proximal tubules High; general IHC practice). | Identify proximal tubules on the counterstained section and check a separate named positive tissue in the same run (HPA: High-staining cells; general IHC practice). |
| IF/ICC shows Golgi signal, while paraffin IHC is being scored for cytoplasmic and membranous staining (HPA subcellular ICC-IF; HPA tissue IHC: profile). | The supplied IF/ICC and tissue IHC records describe different assay observations (HPA subcellular ICC-IF; HPA tissue IHC). | Score the IHC slide using its tissue cell and compartment reference; document IF/ICC localization separately (HPA tissue IHC: profile; HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing APC staining
M00008 · IHCIHC analysis of APC using anti-APC antibody (M00008). APC was detected in a paraffin-embedded section of human colon tissue. Heat mediated a
M00008 · IHCIHC analysis of APC using anti-APC antibody (M00008). APC was detected in a paraffin-embedded section of human colon cancer tissue. Heat med
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000134982-APC/tissue/cerebral+cortex#imid_20391149
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000134982-APC/tissue/colon#imid_20391075
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000134982-APC/tissue/liver#imid_20391060
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000134982-APC/tissue/kidney#imid_20391061
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA013362; green = APC, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA013362; green = APC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for APC IHC & IF
🧪Run appendix first: glandular cells should stain for APC (HPA: High in appendix glandular cells). Run adipose tissue as the negative comparator because adipocytes are not detected (HPA: Not detected in adipocytes); use non-glandular cells on the appendix slide as an internal background reference only after confirming their APC status independently, since the supplied HPA row reports glandular cells only (HPA: appendix row).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show APC in Rh30, SiHa, U2OS, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a rabbit IgG control matched to the primary antibody’s clonality where known, and APC-knockout tissue or cells as a biological negative (caption: rabbit anti-APC; standard IHC practice). For chromogenic appendix sections, quench endogenous peroxidase and inspect gland lumina for DAB background (caption: peroxidase secondary and DAB; standard IHC practice).
⚠️Feasibility: The selected M00008 paraffin-section caption reports EDTA retrieval at pH 8.0 and a 1:50 primary dilution, but its fixative is unreported (M00008 tissue-IHC caption). A target-specific fixation window and fixation effect are unreported in the supplied evidence; the caption describes retrieval, but does not establish that retrieval is required for all APC antibodies (M00008 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; for appendix IHC, assess gland lumina for DAB background before scoring epithelial staining (HPA: High in appendix glandular cells; caption: DAB detection; standard IHC practice).
HPA tissue IHC evidence for APC
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced APC IHC Tips
Troubleshoot APC staining in paraffin sections by checking retrieval, cellular pattern and matched controls before interpreting differences between samples.
Which retrieval condition should I try first for weak APC staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M00008). The human colon IHC example combines this condition with 1:50 primary antibody overnight at 4°C; keep the antibody step constant while comparing adjacent sections (datasheet M00008; standard IHC practice). If staining remains weak, vary heating duration on matched sections, then consider another buffer as a fallback while monitoring tissue morphology and a no-primary control (standard IHC practice). Compare cytoplasmic and membranous signal within the same cell population, since both patterns are reported for APC, and reject conditions that mainly increase diffuse background (HPA tissue IHC; standard IHC practice).
Could fixation explain weak APC staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative, and its staining cannot establish which fixation conditions preserve APC (datasheet M00008). Record fixative, fixation duration and processing history for each sample, then compare similarly processed sections before changing the antibody dilution (standard IHC practice). Use a documented positive control, such as appendix glandular cells, alongside test sections; that cell population has high reported APC staining (HPA tissue IHC). If signal is weak across the run, first repeat the documented EDTA pH 8.0 retrieval and 1:50 overnight primary incubation before attributing failure to fixation (datasheet M00008; standard IHC practice).
Is predominantly cytoplasmic APC staining plausible when membrane signal is faint?
Yes: tissue IHC reports general cytoplasmic and membranous expression, while UniProt places APC at adherens junctions, in the cytoplasm and cytoskeleton, and at cell projections (HPA tissue IHC; UniProt P25054 subcellular). APC has no annotated transmembrane segment, so membrane-associated staining need not outline every cell as a continuous rim (UniProt P25054 topology; standard IHC interpretation). Assess both compartments in morphologically intact cells and compare adjacent sections processed with EDTA pH 8.0 retrieval (datasheet M00008; standard IHC practice). A punctate or edge-only pattern requires control review before assignment to APC, especially if the expected glandular-cell signal is absent (HPA tissue IHC; standard IHC practice).
How should I interpret discordant APC staining across tissues or antibody epitopes?
First establish which APC sequence the antibody recognizes; the supplied IHC caption gives a 1:50 dilution but does not map its epitope (datasheet M00008). APC has 3 listed isoforms, and isoform 1A is especially abundant in brain whereas 1B predominates outside brain, so an epitope shared unevenly among isoforms could alter apparent staining (UniProt P25054 isoforms and tissue specificity). UniProt also lists numerous modified residues, but these annotations alone do not demonstrate that fixation or retrieval masks this antibody's epitope (UniProt P25054 modified residues; datasheet M00008). Compare mapped epitopes, matched processing and cell-specific controls before interpreting discordance as altered APC abundance (standard IHC practice).
How can I check APC localisation by multiplex IF alongside chromogenic IHC?
Use IF as a separate localisation check, pairing APC with a marker for the expected cell population, such as a glandular-cell marker when examining glandular epithelium (HPA tissue IHC; standard IF practice). APC is associated with cytoplasm and the cell membrane and has no transmembrane segment; for an intracellular epitope, optimize gentle permeabilisation while retaining a matched staining control (UniProt P25054 subcellular and topology; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel where tissue autofluorescence obscures shorter wavelengths, with single-label and unstained controls (standard IF practice). HPA reports plasma-membrane localisation with additional Golgi localisation in ICC/IF; compare that evidence cautiously with the separate EDTA pH 8.0 chromogenic tissue method (HPA subcellular; datasheet M00008).
What should I change when APC IHC shows diffuse brown background?
Run no-primary and secondary-only controls to distinguish detection-system signal from staining associated with the primary antibody (standard IHC practice). For HRP and DAB detection, check the endogenous peroxidase block, serum blocking, wash stringency and chromogen development time; these are general IHC controls, not APC-specific findings (standard IHC practice). The selected colon example used 10% goat serum, 1:50 primary antibody overnight at 4°C, and a 30-minute secondary incubation at 37°C (datasheet M00008). Adjust one step at a time while preserving EDTA pH 8.0 retrieval, and judge improvements by signal in intact glandular cells relative to background (datasheet M00008; HPA tissue IHC; standard IHC practice).
How should I score APC staining across sections with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: APC tissue IHC has general cytoplasmic and membranous expression, and glandular cells in several tissues show high staining (HPA tissue IHC). For a cell-based H-score, assign intensity 0–3 and multiply each intensity by its percentage of viable target cells, yielding 0–300 (standard IHC practice). Alternatively, report percentage positive cells or positive-cell density per mm² of annotated viable target tissue, with cytoplasmic and membranous results recorded separately (standard IHC practice). Normalize to the counted target cells or their viable area, and keep retrieval, imaging and positivity thresholds consistent across sections (standard IHC practice).
How do I distinguish genuine APC staining from an IHC artefact?
Look for reproducible cytoplasmic or membranous staining in intact cells; those distributions fit the tissue IHC profile and APC's annotated cellular locations (HPA tissue IHC; UniProt P25054 subcellular). Check cell identity against the specimen: appendix glandular cells are reported as high, whereas adipocytes are reported as not detected, though either reference is context rather than proof for a new section (HPA tissue IHC). Edge accentuation, necrotic deposits and signal persisting in a no-primary control warrant review for processing or detection artefact (standard IHC practice). With HRP–DAB detection, verify the peroxidase block and compare controls before treating brown signal as APC; the selected IHC example used DAB after EDTA pH 8.0 retrieval (standard IHC practice; datasheet M00008).
Boster reagents
Best APC / Adenomatous polyposis coli protein IHC Antibodies
M00008 has IHC images from paraffin sections of human colon and colon cancer (IHC captions). IF/ICC is listed without an image (catalog). Reactivity includes human, mouse and rat (catalog).
Real IHC data
Anti-APC Monoclonal Antibody
Cat # M00008
M00008 is shown staining paraffin sections of human colon and colon cancer (IHC captions). M00008 lists IHC and IF/ICC applications and human, mouse and rat reactivity; no IF image is supplied (catalog).
Which to pick: Choose M00008 for tissue IHC: its rabbit monoclonal antibody has an image from a paraffin section of human colon (catalog; IHC caption). M00008 also lists IF/ICC and human, mouse and rat reactivity, but the supplied images demonstrate human IHC only (catalog; IHC captions). The IHC captions report EDTA retrieval and a 1:50 primary dilution; the fixative is unreported (IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P25054 (APC_HUMAN, Adenomatous polyposis coli protein).
- Human Protein Atlas. APC tissue IHC expression (reliability: Supported).
- Human Protein Atlas. APC subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the Golgi apparatus..
- Human Protein Atlas. APC antibody validation summary (2 antibodies).
- Molecular and immunohistochemical study of APC exon 16 and its possible role in colorectal carcinoma development. Heliyon 2024 — PMC10865248.
- APC promoter methylation and protein expression in hepatocellular carcinoma. Journal of cancer research and clinical oncology 2008 — PMC2757596.
- Anaphase-Promoting Complex 7 is a Prognostic Factor in Human Colorectal Cancer. Annals of coloproctology 2017 — PMC5603343.
- The C-terminus of Apc does not influence intestinal adenoma development or progression. The Journal of pathology 2012 — PMC4610063.
- PubMed PMID:1678319 — UniProt-cited evidence.
- PubMed PMID:1651562 — UniProt-cited evidence.
- PubMed PMID:10947987 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for APC (Adenomatous polyposis coli protein). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00008). Validated IHC antibody: M00008 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet M00008); 2. Fixation: Image fixative and duration unreported (datasheet M00008); 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 M00008); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00008); 8. Primary antibody: Rabbit monoclonal (clone FAD-1) anti-APC, 1:50 (datasheet M00008); 9. Primary incubation: Overnight at 4 °C (datasheet M00008). Watch for: Fixation: Keep paraffin-section fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Cortex neuropil stains highly; caudate glia are undetected (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: HGF shifts APC toward cell protrusions (UniProt). Evidence: UniProt P25054, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet M00008, PMC2757596, PMC5603343. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/APC/immunohistochemistry-ihc-guide.