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ANAPC2 / Anaphase-promoting complex subunit 2 · IHC design guide
Design Immunohistochemistry for ANAPC2
Use this ANAPC2 paraffin IHC guide to plan chromogenic staining with the catalog antibody (datasheet A06153-1). Rectal glandular cells show high staining, while nuclear signal occurs in subsets of cells across tissues (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ANAPC2 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 | Nuclear staining in subsets of tissue cells (HPA tissue IHC) | |
| Staining pattern | Nuclear signal in subsets of cells across several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06153-1) | |
| 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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining is restricted to cell subsets within tissues (HPA tissue IHC) | |
| Regulation | Cell-cycle activity; stain effect unknown (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage needs checking (UniProt) |
Section 1
Recommended ANAPC2 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A06153-1). The published CD34/ANAPC2 chromogenic IHC protocol below uses citrate retrieval (PMC5609928).
| Sample | Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A06153-1) |
| Fixation | Image fixative and duration unreported (datasheet A06153-1); 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 A06153-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06153-1) |
| Primary antibody | Rabbit anti-ANAPC2, 2-5μg/ml (datasheet A06153-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06153-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06153-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ANAPC2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in a subset of cells of several different tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A06153-1); use citrate pH 6.0 when reproducing the published protocol (PMC5609928).
Section 2
What Is the Expected ANAPC2 Staining Pattern?
ANAPC2 staining in paraffin sections should be predominantly nuclear in a subset of cells, including appendix and colon glandular cells and lymph node germinal center cells (HPA: tissue IHC profile; High in these cell types). The tissue IHC assessment is Enhanced, with medium consistency between staining and RNA expression (HPA: reliability). ANAPC2 has no annotated transmembrane segment (UniProt Q9UJX6 topology).
| Strong nuclear staining in glandular cells of appendix, colon, duodenum or rectum, or germinal center cells of tonsil or lymph node (HPA: High in these cell types). | This fits the reported IHC pattern when staining is confined to a subset of cells (HPA: nuclear expression in a subset of cells). Compare nuclei within the same tissue; a uniformly positive section is not the reported pattern. |
| Predominantly membranous, extracellular or diffuse cytoplasmic staining, with little nuclear signal (HPA: nuclear tissue IHC profile). | Treat this as a compartment mismatch and investigate staining artefact or antibody specificity. The absence of a transmembrane segment does not support a membrane pattern (UniProt Q9UJX6 topology). Mitochondrial signal is an additional ICC-IF observation, not the principal tissue IHC pattern (HPA: subcellular). |
| Prominent staining of adipocytes, cardiomyocytes or skeletal myocytes (HPA: Not detected in these cell types). | Suspect cross-reactivity or endogenous detection activity if these cells are strongly positive, especially without the expected nuclear staining in a control section (HPA: tissue IHC profile; general IHC practice). A reported negative is a comparison point, not proof that every specimen must be blank. |
| Broad haze across nuclei, cytoplasm and tissue spaces, obscuring individual positive cells (HPA: nuclear staining in a subset of cells). | The distribution cannot be scored reliably as ANAPC2. Check background controls, blocking, washes and detection conditions using general IHC practice; diffuse colour alone does not establish where the target is expressed. |
| No nuclear signal in appendix or colon glandular cells, or lymph node germinal center cells (HPA: High in these cell types). | Consider failed detection, inadequate assay sensitivity or specimen variation before calling the target absent (general IHC practice). HPA reports cell-specific staining, so confirm that the expected cell population is present and assess a control section. |
💡Expected ANAPC2 appearanceCall a section positive when a subset of the relevant glandular or germinal center cells shows clear, often strong nuclear staining (HPA: nuclear profile; High in listed cells); broad membranous or diffuse background colour is suspicious (HPA: tissue IHC profile; general IHC practice).
| Tissue and cell selection (HPA: tissue IHC) | HPA reports High staining in specified glandular, hematopoietic and germinal center cells, but Not detected in several other named cell populations (HPA: tissue IHC). Select and read controls by cell type, since a tissue name alone can conceal a mixed population. |
| Antibody evidence (HPA: antibody validation) | The tissue IHC assessment is Enhanced, and CAB018692 has Enhanced IHC validation; HPA066539 has no supplied IHC rating (HPA: antibody list; tissue reliability). Apply the observed tissue pattern to an IHC-validated antibody and evaluate another reagent on its own evidence. |
| IF/ICC: where should ANAPC2 appear? (HPA: subcellular) | Mainly in the nucleoplasm, with an additional mitochondrial location in ICC-IF (HPA: supported nucleoplasm; approved mitochondria). This answers the IF/ICC localisation question; the mitochondrial observation should not override the predominantly nuclear paraffin IHC expectation (HPA: tissue IHC profile). |
| Protein form and epitope (UniProt Q9UJX6) | UniProt lists 2 isoforms and no signal peptide, propeptide or transmembrane segment (UniProt Q9UJX6). The supplied record gives no antibody epitope, so isoform coverage and epitope-dependent staining cannot be predicted from it. |
| Detection chemistry (general IHC practice) | Endogenous enzyme activity and nonspecific reagent binding can add chromogenic colour (general IHC practice). Review controls before interpreting colour in an HPA Not detected cell type as ANAPC2; this is a general assay consideration, not an ANAPC2-specific property. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank (HPA: High in appendix and colon glandular cells). | The relevant cells may be absent from the section, or the assay may lack detectable signal (general IHC practice). | Verify the cell population on the counterstain, then check an appropriate positive control and the IHC-validated antibody's documented conditions (HPA: tissue IHC; general IHC practice). Do not infer ANAPC2 fixation sensitivity from this result. |
| Colour appears mainly at membranes or outside cells (HPA: nuclear tissue IHC profile). | This differs from the reported nuclear distribution and may reflect background or nonspecific binding (HPA: tissue IHC; general IHC practice). | Compare with a negative detection control and inspect nuclei in a known-positive cell population. Reassess antibody specificity before scoring this colour as ANAPC2 (general IHC practice). |
| Adipocytes or cardiomyocytes stain strongly (HPA: Not detected in these cell types). | Cross-reactivity or endogenous detection activity is possible; the HPA pattern does not support these cells as routine positive controls (HPA: tissue IHC; general IHC practice). | Examine a control omitting primary antibody and compare the suspect cells with an expected nuclear-positive population in a separate control section (HPA: tissue IHC; general IHC practice). |
| Diffuse colour hides the boundary between positive and negative cells (HPA: subset nuclear profile). | Excess background may come from nonspecific binding, incomplete washing or detection chemistry (general IHC practice). | Check negative controls and review blocking, washes and detection settings. Score only discrete nuclear staining in identifiable cells (general IHC practice; HPA: tissue IHC profile). |
| Every cell is called positive despite visibly mixed staining (HPA: nuclear expression in a subset of cells). | A tissue-wide score can miss the reported cell-specific pattern; HPA also reports low tissue RNA specificity (HPA: tissue IHC and RNA specificity). | Record the stained cell type, nuclear localisation and fraction of cells rather than assigning one score to the whole tissue (HPA: tissue IHC profile; general IHC practice). |
| Mitochondrial-looking IF signal is treated as the expected paraffin IHC result (HPA: subcellular ICC-IF). | The additional mitochondrial location comes from ICC-IF, while tissue IHC reports nuclear expression in a subset of cells (HPA: subcellular; tissue IHC). | Interpret each application against its own HPA observation: assess nucleoplasm and possible mitochondria in IF/ICC, and cell-specific nuclear staining in paraffin IHC (HPA: subcellular; tissue IHC). |
Experiment evidence
IHC & IF images showing ANAPC2 staining
A06153-1 · IHCIHC analysis of ANAPC2 using anti-ANAPC2 antibody (A06153-1). ANAPC2 was detected in paraffin-embedded section of human rectal cancer tissue
A06153-1 · IFIF analysis of ANAPC2 using anti-ANAPC2 antibody (A06153-1). ANAPC2 was detected in immunocytochemical section of A431 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000176248-ANAPC2/tissue/cerebral+cortex#imid_4649808
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000176248-ANAPC2/tissue/colon#imid_4649742
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000176248-ANAPC2/tissue/liver#imid_4649743
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000176248-ANAPC2/tissue/kidney#imid_4649736
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB018692; green = ANAPC2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB018692; green = ANAPC2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ANAPC2 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); on the colon slide, assess morphologically distinct cells for absent or background-level staining, without presuming a particular cell type is ANAPC2-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 ANAPC2 in A-431, U-251MG, U2OS, HaCaT, MCF-7, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control (selected-SKU tissue-IHC caption: rabbit primary antibody; clonality unreported), and ANAPC2-knockout material as a biological negative. Block endogenous peroxidase and assess endogenous biotin background when using the caption’s biotin–streptavidin/DAB detection system (selected-SKU tissue-IHC caption: biotinylated secondary, streptavidin–biotin complex, DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not state a fixative (selected-SKU tissue-IHC caption: fixative not stated). The caption reports heat-mediated retrieval in EDTA at pH 8.0 for a paraffin section, but does not establish how dependent ANAPC2 detection is on retrieval (selected-SKU tissue-IHC caption: heat-mediated EDTA retrieval). ICC-IF images exist, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA subcellular: ICC-IF images); check for background from the biotin-based detection system (selected-SKU tissue-IHC caption: streptavidin–biotin detection).
HPA tissue IHC evidence for ANAPC2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ANAPC2 IHC Tips
Troubleshoot ANAPC2 staining in paraffin sections by checking retrieval, nuclear localization, cell type, controls, and scoring before interpreting chromogenic signal.
Which retrieval conditions should I try first for weak ANAPC2 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody (datasheet A06153-1). The reported paraffin-section image used that retrieval before 10% goat-serum blocking and 2 µg/mL primary antibody overnight at 4°C (caption A06153-1). If staining remains weak, compare a modestly longer heating interval on matched sections while keeping detection and exposure to DAB consistent (standard IHC practice). Include a known positive compartment, such as rectal glandular cells, and assess nuclear signal before accepting any increase in diffuse staining as improved retrieval (HPA tissue IHC: rectal glandular cells High; HPA tissue IHC: nuclear expression).
Could fixation explain weak or uneven ANAPC2 staining in paraffin sections?
Target-specific sensitivity to fixation is unknown; the selected paraffin-section caption does not state a fixative (caption A06153-1: fixative not stated). Record the fixative, fixation duration, section thickness, and processing history for every comparison, then stain matched sections together (standard IHC practice). If a batch differs, compare its nuclear signal in a tissue compartment with reported expression, such as colon glandular cells, before changing antibody concentration (HPA tissue IHC: colon glandular cells High). Keep retrieval at EDTA pH 8.0 during that comparison, because it is the condition documented for this antibody; do not assign a fixation effect without controlled evidence (datasheet A06153-1).
How should I evaluate cytoplasmic staining when ANAPC2 appears mainly nuclear?
Score nuclear staining separately because tissue IHC reports expression in nuclei of a subset of cells across several tissues (HPA tissue IHC: nuclear expression). Supported nucleoplasmic localization and an additional mitochondrial location come from subcellular imaging, while UniProt supplies no subcellular annotation (HPA subcellular: Nucleoplasm supported; HPA subcellular: Mitochondria approved; UniProt Q9UJX6: subcellular location not annotated). Therefore, investigate reproducible punctate cytoplasmic signal, but do not score broad cytoplasmic DAB deposits as equivalent to nuclear positivity. Compare morphology, matched controls, and staining across 2 independently processed sections before attributing an unexpected compartment to ANAPC2 (standard IHC practice).
Can this staining distinguish ANAPC2 isoforms or phosphorylation states?
Do not assign isoform specificity from staining alone: the record lists 2 isoforms, but the supplied antibody caption does not map its epitope (UniProt Q9UJX6: isoforms 1 and 2; caption A06153-1). ANAPC2 has no annotated transmembrane segment, and its listed modifications include phosphoserines at positions 218, 314, 470, 534, and 697 (UniProt Q9UJX6: topology and modified residues). Those annotations do not establish whether fixation or retrieval masks this antibody’s binding site. For an isoform or phosphorylation claim, require an epitope-mapped reagent and an appropriate perturbation or independently validated comparison; report ordinary staining as ANAPC2 immunoreactivity (standard IHC practice).
How can I check ANAPC2 localization by IF alongside the paraffin IHC result?
Use a nuclear counterstain and multiplex with a marker that identifies the expected glandular cell population, then compare cell-level localization with the paraffin-section result (HPA tissue IHC: rectal glandular cells High; HPA tissue IHC: nuclear expression). Choose a fluorophore channel with low measured tissue autofluorescence and include unstained and single-stain controls before interpreting faint signal (standard IF practice). Permeabilize sufficiently to expose an intracellular nuclear epitope; ANAPC2 has no annotated transmembrane segment, but this antibody’s epitope is not mapped in the supplied caption (UniProt Q9UJX6: topology; caption A06153-1). The reported 2 µg/mL overnight condition belongs to paraffin chromogenic IHC, so optimize IF staining separately (caption A06153-1).
What should I change when DAB background obscures ANAPC2-positive nuclei?
First inspect no-primary and secondary-only controls for staining that persists without ANAPC2 antibody (standard IHC practice). The reported workflow uses biotinylated goat anti-rabbit secondary antibody, a streptavidin-biotin complex, and DAB, so assess background from that detection chain on matched sections (caption A06153-1). Apply a peroxidase block and check whether endogenous biotin requires blocking or a different detection system; these are general chromogenic IHC controls, not evidence of ANAPC2 expression (standard IHC practice). Preserve EDTA pH 8.0 retrieval and the reported 10% goat-serum block initially, then adjust antibody concentration or wash stringency one variable at a time (datasheet A06153-1; caption A06153-1).
How should I quantify ANAPC2 staining across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and analysis regions before viewing outcomes, then score nuclear positivity separately from cytoplasmic deposits (HPA tissue IHC: nuclear expression; standard IHC practice). Report an H-score from staining intensity and percentage of positive target cells, or report percentage positive using a fixed threshold; use positive nuclei per mm² when cell density varies (standard IHC practice). Normalize counts to evaluable cells of the same annotated cell type or to viable tissue area, and exclude necrosis and section edges consistently (standard IHC practice). Keep staining batches, imaging settings, and thresholds aligned; the HPA profile describes nuclear expression in a subset of cells, making whole-section averages sensitive to cellular composition (HPA tissue IHC: nuclear expression).
Which findings support true ANAPC2 signal rather than staining artefact?
Give greatest weight to reproducible nuclear staining in an expected cell population, such as rectal glandular cells, with clean no-primary controls (HPA tissue IHC: rectal glandular cells High; HPA tissue IHC: nuclear expression; standard IHC practice). Treat staining confined to section edges, necrotic areas, or areas positive in detection-only controls as suspect (standard IHC practice). Diffuse signal restricted to adipocytes or cardiomyocytes warrants particular scrutiny because those cells were reported as not detected in the tissue profile (HPA tissue IHC: adipocytes Not detected; HPA tissue IHC: cardiomyocytes Not detected). ANAPC2 is an APC/C component, but a positive DAB stain alone does not measure APC/C activity or cell-cycle phase (UniProt Q9UJX6: function; standard IHC practice).
Boster reagents
Best ANAPC2 / Anaphase-promoting complex subunit 2 IHC Antibodies
The catalog antibody A06153-1 has IHC data from a human paraffin section and IF data from A431 cells (IHC/IF captions). Listed reactivity includes human, monkey, mouse, and rat (catalog).
Real IHC data
Anti-ANAPC2 Antibody ®
Cat # A06153-1
A06153-1 will render with an IHC image from a paraffin section of human rectal cancer tissue (IHC caption). The same SKU supports IF/ICC and has an IF image from A431 cells (catalog applications; IF caption).
Which to pick: For tissue IHC, choose A06153-1 at 2–5 μg/mL for human paraffin sections (catalog IHC dilution; IHC caption); the fixative is unreported (IHC caption). For IF/ICC, the same SKU is listed at 5 μg/mL for human samples, with IF shown in A431 cells (catalog applications and IF dilution; IF caption). For cross-species work, this SKU lists monkey, mouse, and rat reactivity, although the supplied IHC and IF images show human samples only (catalog reactivity; IHC/IF captions); clonality is unreported (catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UJX6 (ANC2_HUMAN, Anaphase-promoting complex subunit 2).
- Human Protein Atlas. ANAPC2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ANAPC2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the mitochondria..
- Human Protein Atlas. ANAPC2 antibody validation summary (2 antibodies).
- Combination of artesunate and WNT974 induces KRAS protein degradation by upregulating E3 ligase ANACP2 and β-TrCP in the ubiquitin-proteasome pathway. Cell communication and signaling : CCS 2022 — PMC8934478.
- APC/C is essential for hematopoiesis and impaired in aplastic anemia. Oncotarget 2017 — PMC5609928.
- PubMed PMID:9469815 — UniProt-cited evidence.
- PubMed PMID:15164053 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ANAPC2 (Anaphase-promoting complex subunit 2). Expected localisation Nuclear staining in subsets of tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06153-1). Validated IHC antibody: A06153-1 at 2-5μg/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 rectal cancer tissue; fixative not specified (datasheet A06153-1); 2. Fixation: Image fixative and duration unreported (datasheet A06153-1); 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 A06153-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06153-1); 8. Primary antibody: Rabbit anti-ANAPC2, 2-5μg/ml (datasheet A06153-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06153-1). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining is restricted to cell subsets within tissues (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage needs checking (UniProt); Regulation: Cell-cycle activity; stain effect unknown (UniProt). Evidence: UniProt Q9UJX6, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06153-1, PMC5609928. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ANAPC2/immunohistochemistry-ihc-guide.