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ANAPC1 / Anaphase-promoting complex subunit 1 · IHC design guide
Design Immunohistochemistry for ANAPC1
Plan chromogenic ANAPC1 IHC on paraffin sections using the catalog antibody at 2–5 μg/mL (datasheet A03471-2). Assess cytoplasmic staining in gastrointestinal glandular cells, while accounting for low consistency between staining and RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ANAPC1 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 staining in GI glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03471-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across tissue sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA data show low consistency (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; one full-length chain (UniProt) |
Section 1
Recommended ANAPC1 IHC & IF Protocols
The catalog antibody protocol (datasheet A03471-2) is accompanied by published ANAPC1 IHC protocols for lung squamous cell carcinoma (PMC11951694) and hepatocellular carcinoma tissue (PMC12142242).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A03471-2) |
| Fixation | Image fixative and duration unreported (datasheet A03471-2); 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 A03471-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03471-2) |
| Primary antibody | Rabbit anti-ANAPC1, 2-5 μg/ml (datasheet A03471-2) |
| Primary incubation | Overnight at 4 °C (datasheet A03471-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03471-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANAPC1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissue types, most abundant in gastrointestinal tract. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A03471-2); the lung tissue protocol also specifies EDTA retrieval (PMC11951694).
Section 2
What Is the Expected ANAPC1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic ANAPC1 staining, especially in gastrointestinal glandular cells (HPA tissue IHC). HPA rates the tissue staining “Approved” but reports low consistency with RNA expression and pending external verification (HPA tissue IHC). ANAPC1 has no transmembrane segment; UniProt provides no subcellular location annotation (UniProt Q9H1A4 topology and subcellular record).
| Cytoplasmic staining in colon, duodenum, or rectal glandular cells. | This fits the reported high staining in those cells (HPA tissue IHC). Compare intensity and distribution with controls on the same run; the HPA pattern has low consistency with RNA expression (HPA tissue IHC). |
| Strong nuclear-only or sharply membrane-bound staining in otherwise expected cells. | This departs from the reported cytoplasmic tissue pattern (HPA tissue IHC). Check antibody specificity and detection controls before scoring it as ANAPC1; UniProt does not assign ANAPC1 a nuclear location or transmembrane segment (UniProt Q9H1A4). |
| Strong staining in lung alveolar cells or adipocytes. | HPA reports ANAPC1 as not detected in these cells (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity, while allowing for sample and assay differences; absence in HPA is not a universal negative standard. |
| Diffuse colour across tissue, including spaces or many unrelated cell types. | A diffuse pattern obscures the expected cell-associated cytoplasmic signal (HPA tissue IHC). Review blocking, washing, chromogen exposure, and reagent-only controls as general IHC troubleshooting steps (general IHC practice). |
| No staining in a section expected to contain positive glandular cells. | Colon, duodenum, and rectum offer reported high-staining cells (HPA tissue IHC). A blank result warrants a same-run positive control and checks of primary antibody, retrieval, and detection before concluding ANAPC1 is absent (general IHC practice). |
💡Expected ANAPC1 appearanceA convincing positive is cell-associated high cytoplasmic staining in gastrointestinal glandular cells (HPA tissue IHC); widespread diffuse colour or strong staining in reported negative cell types calls for control-based review (HPA tissue IHC; general IHC practice).
| Strength and limits of tissue evidence | High staining is reported in appendix, colon, duodenum, gallbladder, and rectal glandular cells, and in several other epithelial cell groups (HPA tissue IHC). The tissue assessment is Approved, with low consistency against RNA and external verification pending (HPA tissue IHC). |
| Choice of positive and low-signal comparators | Colon glandular cells are a reported high-staining comparator; lung alveolar cells and adipocytes are reported as not detected (HPA tissue IHC). These are observed patterns, not guarantees for every specimen or antibody (HPA tissue IHC reliability). |
| Does the IF pattern define the IHC result? | No. ICC-IF reports approved vesicle localisation, with images in A-431, U2OS, and NIH 3T3 cells (HPA subcellular ICC-IF). Tissue IHC reports broader cytoplasmic staining (HPA tissue IHC); score the paraffin section against its IHC pattern. |
| Antibody validation | Three listed rabbit polyclonal antibodies have Approved IHC status; only HPA042998 also has Approved ICC status (HPA antibodies). An Approved label does not remove the tissue-level low-consistency caveat (HPA tissue IHC). |
| Topology and processing | ANAPC1 has no annotated transmembrane segment, signal peptide, propeptide, or separate processed chain (UniProt Q9H1A4). These annotations give no basis to expect a membrane rim or a secreted staining pattern; they do not establish retrieval conditions. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected gastrointestinal glandular cells are blank. | The run may have failed, or staining may vary across samples; HPA reports high staining in these cells but notes limited RNA agreement (HPA tissue IHC). | Check a same-run positive section, primary antibody and detection reagents; then review retrieval using the assay's established controls (general IHC practice). |
| Nuclei dominate while cytoplasm is faint. | The distribution differs from the reported tissue cytoplasmic pattern (HPA tissue IHC); its cause cannot be assigned from localisation alone. | Compare primary-omission and positive controls, inspect counterstain and chromogen separately, and reassess antibody specificity (general IHC practice). |
| Alveolar cells or adipocytes stain strongly. | Both are listed as not detected by HPA tissue IHC; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Use appropriate reagent-only controls and evaluate cell-associated signal against a reported high-staining tissue in the same run (general IHC practice; HPA tissue IHC). |
| Colour is widespread and poorly confined to cells. | Background from insufficient blocking, washing, or overdeveloped chromogen can hide the reported cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Review reagent-only controls, blocking and washes; adjust detection development using assay controls (general IHC practice). |
| Only a vesicle-like pattern is treated as positive. | Vesicle localisation comes from ICC-IF, whereas tissue IHC is described as cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). | Score paraffin-section IHC by tissue compartment and cell type; reserve the vesicle observation for the separate IF/ICC assessment (HPA tissue IHC; HPA subcellular ICC-IF). |
| All tissue types show equally intense staining. | That distribution conflicts with HPA's high, low, and not-detected cell groups, though its tissue assessment has limited RNA agreement (HPA tissue IHC). | Compare a reported high-staining glandular sample with reported negative cells; review dilution and detection controls before assigning biological meaning (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ANAPC1 staining
A03471-2 · IHCIHC analysis of APC1/ANAPC1 using anti-APC1/ANAPC1 antibody (A03471-2). APC1/ANAPC1 was detected in a paraffin-embedded section of human pla
A03471-2 · IFIF analysis of APC1/ANAPC1 using anti-APC1/ANAPC1 antibody (A03471-2). APC1/ANAPC1 was detected in an immunocytochemical section of MCF-7 ce
A03471 · IHCImmunohistochemistry of APC1 in human brain tissue with APC1 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000153107-ANAPC1/tissue/cerebral+cortex#imid_9021068
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000153107-ANAPC1/tissue/colon#imid_9013290
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000153107-ANAPC1/tissue/liver#imid_9013299
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000153107-ANAPC1/tissue/pancreas#imid_9013250
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042998; green = ANAPC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042998; green = ANAPC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANAPC1 IHC & IF
🧪Run appendix first and score its glandular cells, which HPA rates High (HPA: appendix glandular cells, High). Use adipose tissue adipocytes as a negative comparator (HPA: adipocytes, Not detected); on the appendix slide, treat unstained nonglandular cells as internal background comparators only after confirming they are unstained.
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 ANAPC1 in A-431, U2OS, NIH 3T3, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, matched to the primary antibody’s clonality where applicable (caption: rabbit primary antibody; standard IHC controls). Use ANAPC1 knockout material as a biological specificity control and quench endogenous peroxidase before HRP/DAB detection, particularly when scoring cells near blood or inflammatory cells (UniProt: ANAPC1; caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A03471-2 paraffin-section caption does not state a fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but it does not establish that this retrieval is required (caption: EDTA retrieval, pH 8.0). Neither frozen sections nor IF/ICC is established as easier by the supplied evidence; for appendix glandular scoring, exclude luminal material from cellular DAB scoring (HPA: appendix glandular cells, High; standard IHC practice).
HPA tissue IHC evidence for ANAPC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced ANAPC1 IHC Tips
Use the catalog antibody’s paraffin section result as the starting point, then check cell type, staining pattern, and controls when interpreting ANAPC1 signal.
What retrieval should I try when ANAPC1 staining is weak in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 (datasheet A03471-2). The catalog antibody detected ANAPC1 in a paraffin embedded human placenta section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A03471-2). Keep section thickness, heating time, cooling, and detection conditions consistent while comparing retrieval runs, so a change in signal has a clear cause (standard IHC practice). If staining remains weak, test an alternative retrieval buffer only as a documented fallback, with a matched positive control and an inspection for tissue damage (standard IHC practice).
Could fixation explain weak ANAPC1 staining in my paraffin sections?
Target specific fixation sensitivity is unknown: the selected tissue IHC caption identifies a paraffin embedded section but does not state its fixative (datasheet A03471-2). Record the fixative and fixation duration for each specimen, then compare sections processed together using the same EDTA pH 8.0 retrieval and antibody conditions (datasheet A03471-2; standard IHC practice). If a well preserved control stains but a test section does not, assess tissue preservation, processing history, and retrieval before assigning a biological difference (standard IHC practice). Do not infer fixation tolerance from ANAPC1 phosphorylation or lack of a transmembrane segment (UniProt Q9H1A4 topology and modified residues).
How should I interpret cytoplasmic and punctate ANAPC1 staining?
Assess the stained compartment and cell type separately: tissue IHC reports cytoplasmic ANAPC1 across several tissues, especially the gastrointestinal tract (HPA tissue IHC). Approved subcellular imaging reports vesicular localisation, so punctate signal can be compared with that observation without treating it as a required paraffin section pattern (HPA subcellular). UniProt does not annotate a subcellular location for Q9H1A4, and its topology lists no transmembrane segment (UniProt Q9H1A4 subcellular and topology). Compare the pattern with a matched positive tissue, a no primary control, and intact cellular morphology before calling isolated deposits specific (HPA tissue IHC; standard IHC practice).
Could ANAPC1 isoforms or epitope masking explain uneven staining?
The supplied UniProt record annotates 0 isoforms, while the selected antibody caption does not identify its binding epitope (UniProt Q9H1A4 isoforms; datasheet A03471-2). That record also lists phosphorylated residues, including serines 51, 60, and 202, but does not establish whether they affect this antibody’s staining (UniProt Q9H1A4 modified residues; datasheet A03471-2). Treat uneven signal as an open technical and biological question rather than assigning it to a particular isoform or modification (standard IHC practice). Compare matched sections under the documented EDTA pH 8.0 retrieval, and seek epitope information before designing a modification specific interpretation (datasheet A03471-2; standard IHC practice).
How can IF help resolve which cells contain ANAPC1 signal?
For a separate IF experiment, multiplex ANAPC1 with a marker identifying the cell population under study, then score colocalisation at the single cell level (standard IF practice). Choose fluorophores and imaging channels after checking tissue autofluorescence, and include single stain controls to distinguish spectral bleed through from shared signal (standard IF practice). ANAPC1 has no annotated transmembrane segment, but the antibody’s epitope is unspecified; optimise permeabilisation for access to the observed intracellular compartment rather than assuming a membrane facing epitope (UniProt Q9H1A4 topology; datasheet A03471-2; standard IF practice). Vesicular IF localisation is reported, while tissue IHC describes cytoplasmic staining, so compare patterns cautiously across preparations (HPA subcellular; HPA tissue IHC).
What should I check when ANAPC1 DAB staining appears diffuse or widespread?
First compare the test section with a no primary control and inspect whether DAB follows cells or accumulates at folds, edges, and damaged areas (standard IHC practice). The catalog result used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and peroxidase based DAB detection (datasheet A03471-2). Include a peroxidase blocking step and review secondary antibody specificity and wash conditions when background persists; these are general chromogenic IHC controls (standard IHC practice). Widespread staining alone is insufficient evidence of ANAPC1, because the tissue IHC profile includes both strongly stained cell populations and populations with no detected signal (HPA tissue IHC).
How should I score ANAPC1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use a consistent intensity scale to calculate an H score or report the percentage of positive cells (standard IHC practice). For spatial comparisons, count positive cells per mm² of viable tissue and normalise to the number of eligible cells or measured viable area (standard IHC practice). Keep retrieval, antibody concentration, DAB development, imaging settings, and positivity thresholds constant across the series (standard IHC practice). Report cell type alongside the score: HPA describes high staining in colon glandular cells but no detected staining in lung alveolar cells, and its tissue IHC reliability note cites low RNA consistency (HPA tissue IHC).
When is ANAPC1 staining convincing rather than an artefact?
A convincing call combines reproducible cell associated staining, an appropriate positive control, and a clean no primary control under matched conditions (standard IHC practice). Compare the stained cell type with the reported tissue pattern: HPA lists high signal in appendix glandular cells and no detected signal in adipocytes, while describing the overall pattern as cytoplasmic (HPA tissue IHC). Treat edge staining, necrotic regions, and deposits present in the no primary control as suspect, and check endogenous peroxidase when interpreting DAB (standard IHC practice). Vesicular IF localisation offers context but does not by itself validate a punctate DAB deposit; HPA also flags low consistency between antibody staining and RNA expression (HPA subcellular; HPA tissue IHC).
Boster reagents
Best ANAPC1 / Anaphase-promoting complex subunit 1 IHC Antibodies
Two anti-ANAPC1 antibodies have human tissue IHC images; one also has cell IF data (catalog images). A03471 lists mouse reactivity, although its pictured IHC uses human tissue (catalog reactivity and IHC caption).
Real IHC data
Anti-APC1/ANAPC1 Antibody ®
Cat # A03471-2
Real IHC data
Anti-APC1 ANAPC1 Antibody
Cat # A03471
A03471-2 has IHC data from a paraffin-embedded human placenta section and IF data from MCF-7 cells (catalog IHC and IF captions). A03471 has IHC data from human brain tissue and lists IHC-P as an application (catalog IHC caption and applications).
Which to pick: For tissue IHC, A03471-2 has a documented paraffin-section workflow with EDTA pH 8.0 retrieval; A03471 has a human brain IHC image, but its caption does not report processing, and neither IHC caption reports the fixative (catalog IHC captions). Choose A03471-2 for IF/ICC because those applications and an MCF-7 IF image are listed for it (catalog applications and IF caption). For mouse-reactive IHC-P, A03471 lists mouse reactivity, but its pictured IHC uses human tissue (catalog reactivity, applications and IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H1A4 (APC1_HUMAN, Anaphase-promoting complex subunit 1).
- Human Protein Atlas. ANAPC1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ANAPC1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. ANAPC1 antibody validation summary (3 antibodies).
- The Potential Biological Roles and Clinical Significance of Anaphase-Promoting Complex Subunit 1 in Colorectal Cancer. Cancer control : journal of the Moffitt Cancer Center 2025 — PMC12033653.
- Elevated expression of ANAPC1 in lung squamous cell carcinoma: clinical implications and mechanisms. Future science OA 2025 — PMC11951694.
- Exploring the potential function of high expression of ANAPC1 in regulating ubiquitination in hepatocellular carcinoma. World journal of gastrointestinal oncology 2025 — PMC12142242.
- PubMed PMID:11179667 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANAPC1 (Anaphase-promoting complex subunit 1). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03471-2). Validated IHC antibody: A03471-2 at 2-5 μg/ml (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 placenta tissue; fixative not specified (datasheet A03471-2); 2. Fixation: Image fixative and duration unreported (datasheet A03471-2); 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 A03471-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03471-2); 8. Primary antibody: Rabbit anti-ANAPC1, 2-5 μg/ml (datasheet A03471-2); 9. Primary incubation: Overnight at 4 °C (datasheet A03471-2). Watch for: Fixation: Keep fixation consistent across tissue sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA data show low consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one full-length chain (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q9H1A4, HPA tissue IHC · Approved, HPA ICC-IF · vesicles., Datasheet A03471-2, PMC11951694, PMC12142242. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANAPC1/immunohistochemistry-ihc-guide.