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APC / Adenomatous polyposis coli protein · IHC design guide
Design Immunohistochemistry for APC
Plan APC chromogenic IHC in paraffin sections using the catalog antibody’s colon section workflow (datasheet M00008). Assess cytoplasmic and membranous staining against cell specific tissue controls (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 | Cytoplasmic and membranous; glandular cells often high (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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium agreement with RNA expression (HPA tissue IHC) | |
| Regulation | Isoform 1A is brain enriched (UniProt) | |
| Isoform / epitope | 3 isoforms; antibody epitope coverage is unreported (UniProt; datasheet M00008) |
Section 1
Recommended APC IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M00008) is accompanied by published APC staining methods for colorectal carcinoma tissue (PMC10865248) and liver tissue (PMC2757596).
| 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 for the catalog antibody (datasheet M00008). Neither included article specifies an antigen-retrieval step (PMC10865248; PMC2757596).
Section 2
What Is the Expected APC Staining Pattern?
In paraffin-section IHC, expect APC staining mainly in the cytoplasm and along cell membranes, including epithelial junctions (HPA: general cytoplasmic and membranous expression; UniProt P25054: adherens junction and cytoplasm). Glandular and respiratory epithelial cells are useful positive populations (HPA: High in appendix glandular cells and bronchus respiratory epithelial cells). APC has no transmembrane segment, so membrane-associated staining does not imply a membrane-spanning protein (UniProt P25054 topology). HPA rates tissue IHC reliability as Supported, with medium consistency against RNA expression (HPA tissue IHC).
| Cytoplasmic staining with a membranous outline in glandular epithelium. | This fits APC's reported tissue pattern (HPA: general cytoplasmic and membranous expression). Appendix, breast, duodenum and gallbladder glandular cells each provide reported High examples (HPA tissue IHC). Judge the pattern within the identified cells; surrounding cells need not share their intensity. |
| Predominantly nuclear staining, with little cytoplasmic or membranous signal. | That is outside the supplied dominant IHC pattern (HPA: general cytoplasmic and membranous expression) and listed UniProt locations (UniProt P25054 subcellular). Treat it as unconfirmed staining; check the same run against a reported positive cell population and the detection controls before assigning it to APC. |
| Strong staining in adipocytes, cardiomyocytes or ovarian follicle cells. | These particular populations were Not detected in the HPA survey (HPA: adipose adipocytes, heart cardiomyocytes, ovary follicle cells). Reproducible signal there merits checks for antibody cross-reactivity or endogenous detection activity. The HPA observations concern those cells, not every cell in each tissue. |
| Diffuse color across tissue, lumens or a section with no primary antibody. | A pattern without cell-specific cytoplasmic or membranous enrichment is less convincing than the reported APC IHC profile (HPA: general cytoplasmic and membranous expression). In a no-primary control, color points to detection background, including possible endogenous activity; this is a general chromogenic IHC control interpretation. |
| No staining in the expected positive cells of a control section. | A blank result in appendix glandular cells or bronchial respiratory epithelium conflicts with their reported High staining (HPA tissue IHC). First verify that the relevant cells are present and the control run worked. HPA's Supported rating still reflects only medium antibody–RNA consistency, so a single negative slide does not establish biological absence (HPA tissue IHC). |
💡Expected APC appearanceCall a result positive when identified glandular or respiratory epithelial cells show clear cytoplasmic and membranous staining, potentially strong in reported High populations (HPA tissue IHC); isolated nuclear color or strong signal in reported Not detected cell types requires control review (UniProt P25054 subcellular; HPA tissue IHC).
| Tissue and cell selection | Use a named positive cell population when judging the run: appendix glandular cells, bronchial respiratory epithelium, kidney proximal tubule cell bodies and liver cholangiocytes are High (HPA tissue IHC). HPA's Not detected calls are cell-specific and should not be used as whole-organ absence claims (HPA tissue IHC). |
| Compartment and cellular state | UniProt places APC at adherens junctions, the cytoskeleton, lamellipodia and ruffle membranes (UniProt P25054 subcellular). Those locations support some pattern variation, while HPA's tissue-level expectation remains broadly cytoplasmic and membranous (HPA tissue IHC). HGF-associated projection localization is context-dependent, not a required paraffin-section feature (UniProt P25054 subcellular). |
| Isoforms and antibody recognition | UniProt lists isoforms 1A, 2 and 1B; 1A is especially strong in brain, while 1B predominates outside brain (UniProt P25054 tissue specificity). The supplied sources do not identify the catalog antibody's epitope or isoform coverage, so do not infer isoform-specific staining from an IHC result. |
| Evidence and antibody validation | HPA labels tissue IHC Supported with medium staining–RNA consistency (HPA tissue IHC). Its listed rabbit polyclonal HPA013349 has IHC Supported status; HPA013362 has ICC Supported status, with no IHC status supplied (HPA antibodies). These labels support the reported applications but do not establish identical performance for an unspecified catalog antibody. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive control is blank. | The relevant reported High cell population may be absent, or a general IHC workflow step may have failed (HPA: High in appendix glandular cells). | Confirm the cell population on the section, then review retrieval, primary-antibody use and chromogenic detection against the laboratory's validated IHC workflow (general IHC practice). Target-specific fixation sensitivity is unreported in the supplied sources. |
| All cells have similar diffuse color. | Non-specific staining or detection background may obscure the cell-linked cytoplasmic and membranous pattern (HPA tissue IHC; general IHC practice). | Compare with a no-primary control; review blocking, washes and detection conditions using the assay's established controls (general IHC practice). Avoid interpreting uniform color as APC expression. |
| A reported Not detected cell population stains strongly. | Antibody cross-reactivity or endogenous detection activity is possible; HPA reports adipocytes and cardiomyocytes as Not detected (HPA tissue IHC). | Check a no-primary control and an independently validated APC antibody if available (general IHC practice). Record the exact stained cell type before comparing it with HPA. |
| Only nuclear staining is apparent. | The location disagrees with the supplied cytoplasmic and membranous tissue profile (HPA tissue IHC) and UniProt's listed APC compartments (UniProt P25054 subcellular). | Compare its distribution with a reported High epithelial control, inspect the no-primary control and withhold an APC-positive call until the compartment discrepancy is resolved (general IHC practice). |
| Two tissues give different staining intensities. | Cell populations differ: kidney proximal tubule cell bodies are High, whereas prostate glandular cells are Low (HPA tissue IHC). Isoform expression also varies by tissue (UniProt P25054 tissue specificity). | Score the specified cell populations separately and compare each with its own HPA entry. Do not treat a lower-intensity population as a failed run when a reported High control performs as expected. |
| Q: What if IF/ICC shows a different APC pattern? | HPA reports mainly plasma-membrane localization and additional Golgi localization in ICC-IF (HPA subcellular); imaging format and cells differ from paraffin tissue IHC. | A: Interpret the IF/ICC image against its own controls and the dedicated IF/ICC guide. Do not use the Golgi finding as a required chromogenic tissue-IHC pattern (HPA subcellular; HPA tissue IHC). |
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 (HPA: High in appendix glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); within the appendix slide, treat cells without specific staining as a background reference, without assuming that all nonglandular cells are APC-negative.
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) and rabbit IgG isotype controls matched to the primary antibody’s clonality, plus an APC-knockout biological negative where available (M00008 caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase and inspect appendix background before interpreting DAB staining (M00008 caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the M00008 paraffin-section caption does not state a fixative (selected-SKU caption). Heat retrieval in EDTA at pH 8.0 was used for the catalog antibody in human colon, but retrieval dependency in appendix is unreported (selected-SKU caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; endogenous peroxidase may contribute to DAB background in appendix (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, cell compartment, antibody epitope, controls, and scoring before interpreting chromogenic signal.
What retrieval should I try first when APC staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet M00008). The selected human colon section used this retrieval before overnight incubation at 4°C with antibody diluted 1:50 (datasheet M00008). If staining remains weak, compare a modest range of heating times while keeping EDTA and pH constant, and assess tissue morphology alongside signal (standard IHC practice). Include a consistently stained control section in each run, since changes in heating, section age, or detection can otherwise masquerade as retrieval effects (standard IHC practice). Score cytoplasmic and membranous compartments separately when comparing conditions (HPA: general cytoplasmic and membranous expression).
How should I troubleshoot variable APC staining when fixation details are missing?
The selected APC caption identifies a paraffin embedded human colon section but does not state its fixative (datasheet M00008). APC specific sensitivity to fixation is therefore unknown from the supplied evidence; neither the tissue staining profile nor protein topology establishes a fixation effect (datasheet M00008; HPA: tissue IHC; UniProt P25054: no transmembrane segment). Record the fixative, time to fixation, fixation duration, and processing history for each specimen before comparing staining intensity (standard IHC practice). Keep retrieval at EDTA pH 8.0 and antibody dilution at 1:50 while assessing fixation related variation against a control section processed in the same run (datasheet M00008; standard IHC practice).
Which APC staining compartments should I expect in chromogenic tissue sections?
Evaluate cytoplasmic and membranous staining as plausible APC patterns in tissue sections (HPA: general cytoplasmic and membranous expression). APC is also annotated at adherens junctions, the cytoskeleton, lamellipodia, and ruffle membranes, without a transmembrane segment (UniProt P25054: subcellular location and topology). In cell imaging, the plasma membrane is the supported main location and the Golgi apparatus an approved additional location; these observations provide context rather than a required pattern for every paraffin section (HPA: subcellular). Compare staining with cell boundaries and tissue architecture under the same counterstain, and inspect any exclusively nuclear or diffuse extracellular signal for background before scoring (standard IHC practice).
Could antibody epitope placement change which APC isoforms I detect?
Check the catalog antibody's immunogen or mapped epitope before interpreting an absent or altered APC signal; an epitope location is not supplied here (datasheet M00008: epitope not stated). APC has 3 listed isoforms, 1A, 2, and 1B (UniProt P25054: isoforms). Isoform 1A is especially abundant in brain, whereas 1B predominates in other tissues, including gastric mucosa and blood (UniProt P25054: tissue specificity). Compare the antibody's documented epitope with isoform sequences and any known specimen variant before assigning negative staining to loss of all APC forms (standard IHC practice; UniProt P25054: isoforms). Document this uncertainty in the scoring record (standard IHC practice).
How can I check APC localisation by IF alongside the chromogenic IHC result?
Treat IF as a separate assay and establish its own antibody conditions; the selected 1:50 dilution and EDTA pH 8.0 retrieval come from paraffin section IHC (datasheet M00008). Multiplex APC with a validated marker of the expected cell population, such as a glandular cell marker when examining glandular regions, and include single stain controls (HPA: high APC staining in appendix glandular cells; standard IF practice). Choose fluorophores after checking tissue autofluorescence and spectral overlap, with appropriate unstained controls (standard IF practice). For an intracellular APC epitope, optimise permeabilisation after fixation; for a membrane associated epitope, confirm whether access is needed without assuming APC spans the membrane (UniProt P25054: cytoplasmic and membrane localisation, no transmembrane segment; standard IF practice).
What should I adjust when APC DAB staining looks diffuse or nonspecific?
First compare the suspect section with a no primary control and a consistently stained control tissue, checking whether the diffuse signal follows cell outlines or covers the slide indiscriminately (standard IHC practice). The selected colon example used 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, and peroxidase based DAB detection (datasheet M00008). Optimise primary dilution, washing, and blocking one variable at a time while checking that expected cytoplasmic or membranous detail remains visible (HPA: general cytoplasmic and membranous expression; standard IHC practice). Include a peroxidase blocking step and inspect endogenous pigment or damaged tissue before calling persistent brown deposits APC (standard IHC practice).
How should I quantify APC IHC without mixing cell abundance and staining intensity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cell population before scoring, then report the percentage of positive cells and an intensity based H-score using the same thresholds across specimens (standard IHC practice). Record cytoplasmic and membranous scores separately because both patterns occur in tissue IHC (HPA: general cytoplasmic and membranous expression). Normalise positive cell counts or stained area to the number of eligible cells or the measured viable tissue area, expressed per mm² when density is reported (standard IHC practice). Exclude folds, necrosis, and section edges by a prespecified rule, and use a common control section to monitor batch effects (standard IHC practice).
When does brown staining support genuine APC expression rather than artefact?
Give greatest weight to reproducible cytoplasmic or membranous staining in intact, correctly identified cells, with little comparable signal in the no primary control (HPA: general cytoplasmic and membranous expression; standard IHC practice). High staining is reported in appendix glandular cells and kidney proximal tubule cell bodies, while adipocytes are reported as undetected, making cell identity essential to interpretation (HPA: tissue IHC). Investigate exclusively nuclear staining, section edge accentuation, necrotic deposits, and signal attributable to endogenous peroxidase before calling a specimen positive (UniProt P25054: subcellular location; standard IHC practice). Treat weak or absent staining cautiously because HPA rates tissue IHC reliability as Supported with medium agreement between staining and RNA data (HPA: reliability).
Boster reagents
Best APC / Adenomatous polyposis coli protein IHC Antibodies
M00008 has a human colon paraffin-section IHC figure (catalog image caption). IF/ICC and human, mouse and rat reactivity are listed, but no IF image is supplied (catalog applications/reactivity; IF image list).
Real IHC data
Anti-APC Monoclonal Antibody
Cat # M00008
M00008 renders with its human colon paraffin-section IHC figure (catalog image caption). A second caption describes human colon cancer paraffin-section IHC; IF/ICC and mouse/rat reactivity are catalog listings without corresponding images in this payload (catalog image captions; applications/reactivity).
Which to pick: For tissue IHC, choose M00008: its own figure caption documents human colon paraffin-section staining with EDTA retrieval at pH 8.0 and a 1:50 primary dilution (catalog IHC image caption). The fixative is unreported (catalog IHC image caption). For IF/ICC or mouse/rat samples, M00008 is the listed option because it is a rabbit monoclonal clone FAD-1 with those applications and species listed; no IF or mouse/rat IHC image is supplied (catalog host/clone/applications/reactivity; image 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 fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium agreement with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; antibody epitope coverage is unreported (UniProt; datasheet M00008); Regulation: Isoform 1A is brain enriched (UniProt). Evidence: UniProt P25054, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet M00008, PMC10865248, PMC2757596. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/APC/immunohistochemistry-ihc-guide.