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AXIN2 / Axin-2 · IHC design guide
Design Immunohistochemistry for AXIN2
Plan AXIN2 staining in paraffin sections using the catalog antibody’s IHC protocol and the observed cytoplasmic and nuclear tissue pattern (HPA tissue IHC). Compare positive and low-staining cell populations, and interpret intensity with the reported medium consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AXIN2 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 nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular cells can stain; cytoplasmic and nuclear signal (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01772-2) | |
| Positive control | Breast+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A01772-2); verify before use. | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Intensity regulation unreported (UniProt) | |
| Isoform / epitope | 0 listed isoforms; no processing reported (UniProt) |
Section 1
Recommended AXIN2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with three published AXIN2 IHC protocols for rat lung, human ameloblastoma, and osteosarcoma sections (PMC3174135; PMC3934066; PMC8464802).
| Sample | Paraffin-embedded human renal carcinoma tissue; fixative not specified (datasheet A01772-2) |
| Fixation | Image fixative and duration unreported (datasheet A01772-2); 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 A01772-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01772-2) |
| Primary antibody | Rabbit anti-AXIN2, 2-5 μg/ml (datasheet A01772-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01772-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01772-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AXIN2-positive staining in adipocytes of breast (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A01772-2). Citrate retrieval is a published alternative to evaluate (PMC3934066; PMC8464802).
Section 2
What Is the Expected AXIN2 Staining Pattern?
AXIN2 is a cytoplasmic protein with no transmembrane segment (UniProt Q9Y2T1). In paraffin-section IHC, expect cytoplasmic and sometimes nuclear staining in the relevant cells; HPA reports general cytoplasmic and nuclear expression, with high staining in several cell types (HPA tissue IHC). HPA rates its tissue pattern Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining, with or without nuclear staining, in colon or duodenum glandular cells. | This fits HPA's general cytoplasmic and nuclear pattern and its high staining in these glandular cells (HPA tissue IHC). Assess signal within the cells rather than treating the entire gland as uniformly positive. |
| Staining confined to extracellular material, or a sharp membrane-only outline without convincing intracellular signal. | Reassess specificity: AXIN2 has no transmembrane segment and UniProt places it in the cytoplasm (UniProt Q9Y2T1). Membrane staining alone is not established as the expected paraffin-section pattern; HPA's plasma-membrane ICC-IF location is uncertain (HPA subcellular). |
| Strong staining chiefly in a cell population recorded as low, while expected cells show little signal. | Consider nonspecific antibody binding or endogenous detection activity (general IHC practice). HPA records low staining in oral-mucosa and esophageal squamous cells, and in prostate and fallopian-tube glandular cells; low does not mean absent (HPA tissue IHC). |
| A diffuse wash of chromogen obscures cell boundaries across the section. | The distribution cannot support a compartment call. Review blocking, antibody concentration, washes and detection controls (general IHC practice) before comparing it with HPA's cytoplasmic and nuclear pattern (HPA tissue IHC). |
| No detectable signal in a well-preserved colon glandular-cell control. | Investigate the IHC workflow before calling the specimen negative: HPA reports high staining in colon glandular cells (HPA tissue IHC). Check the antibody's IHC-P validation, detection reagents, retrieval conditions and control-section handling (general IHC practice). |
💡Expected AXIN2 appearanceCall a result convincing when distinct cytoplasmic staining, potentially accompanied by nuclear staining, is visible in expected cells such as colon glandular cells or lung macrophages, both reported High by HPA; widespread extracellular color or signal confined to cell outlines warrants investigation (HPA tissue IHC; UniProt Q9Y2T1; general IHC practice).
| Compartment evidence | UniProt lists cytoplasm, while HPA tissue IHC reports general cytoplasmic and nuclear expression (UniProt Q9Y2T1; HPA tissue IHC). Score the observed compartments separately; nuclear color is compatible with HPA tissue staining, but a nucleus-only result needs controls. |
| Choice of positive tissue | HPA reports High staining in breast adipocytes, cerebral-cortex neurons, colon and duodenum glandular cells, lung macrophages and placental trophoblasts (HPA tissue IHC). Choose a control whose relevant cell population is identifiable on the section. |
| Interpretation of low-staining tissue | HPA lists low staining in oral mucosa, esophagus, prostate and fallopian tube, and reports low tissue RNA specificity overall (HPA tissue IHC). A weak signal in those sites is plausible; they are not validated blank controls. |
| Antibody-validation limit | HPA lists CAB012283 and CAB017783 as Approved for IHC; neither is listed as IHC Enhanced in the supplied record (HPA antibodies). Treat agreement with the tissue pattern as supporting evidence, while checking controls for the antibody actually used. |
| IF/ICC cross-check: should its location dictate the IHC call? | No. HPA ICC-IF supports cytosol but labels nucleoplasm and plasma membrane uncertain (HPA subcellular). Use those images as context; judge paraffin-section staining against the tissue IHC pattern and its controls (HPA tissue IHC; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive tissue has no signal. | A failed staining or detection step is possible; HPA reports High staining in colon glandular cells (HPA tissue IHC). | Run a known-positive section alongside the sample; verify antibody suitability for IHC-P, retrieval settings and the detection system (general IHC practice). |
| Most cells show uniform brown haze. | Excess antibody, inadequate washing or insufficient blocking can raise background (general IHC practice). | Compare a no-primary control; adjust antibody concentration, blocking and washes, then reassess cell-level localization (general IHC practice). |
| Color appears mainly in blood-rich or inflammatory areas. | Endogenous detection activity or nonspecific binding can mimic target staining (general IHC practice). | Check the no-primary control and use blocking appropriate to the detection chemistry; evaluate lung macrophages as cells, since HPA reports them High (HPA tissue IHC; general IHC practice). |
| Only nuclei are stained, with little cytoplasmic signal. | HPA tissue IHC allows nuclear expression, but UniProt lists cytoplasm and HPA ICC-IF marks nucleoplasm uncertain (HPA tissue IHC; UniProt Q9Y2T1; HPA subcellular). | Confirm that nuclear color follows recognizable cells in a positive control; review counterstain and no-primary control before accepting a nucleus-only interpretation (general IHC practice). |
| Strong color appears only in a low-staining reference tissue. | Nonspecific signal is possible, but HPA's Low category does not establish absence of AXIN2 (HPA tissue IHC). | Compare the same run with a High reference cell population and a no-primary control; report the observed cell type and compartment rather than calling the low tissue negative (HPA tissue IHC; general IHC practice). |
| A dominant membrane outline conflicts with the expected intracellular pattern. | UniProt places AXIN2 in cytoplasm without a transmembrane segment; HPA's ICC-IF membrane assignment is uncertain (UniProt Q9Y2T1; HPA subcellular). | Inspect a no-primary control and an IHC positive section, and check whether any cytoplasmic or nuclear signal accompanies the outline (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing AXIN2 staining
A01772-2 · IHCIHC analysis of Axin 2/AXIN2 using anti-Axin 2/AXIN2 antibody (A01772-2). Axin 2/AXIN2 was detected in a paraffin-embedded section of human
A01772-2 · IHCIHC analysis of Axin 2/AXIN2 using anti-Axin 2/AXIN2 antibody (A01772-2). Axin 2/AXIN2 was detected in a paraffin-embedded section of human
A01772 · IHCImmunohistochemistry of AXIN2 in rat lung tissue with AXIN2 antibody at 5 μg/mL.
A01772 · IHCImmunohistochemistry of AXIN2 in mouse lung tissue with AXIN2 antibody at 5 μg/mL.
A01772 · IFImmunofluorescence of AXIN2 in Rat Lung cells with AXIN2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000168646-AXIN2/tissue/cerebral+cortex#imid_4510428
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000168646-AXIN2/tissue/colon#imid_4510362
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000168646-AXIN2/tissue/liver#imid_4510355
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000168646-AXIN2/tissue/kidney#imid_4510364
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017783; green = AXIN2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017783; green = AXIN2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AXIN2 IHC & IF
🧪Run colon first and look for staining in glandular cells (HPA: High in colon glandular cells). HPA lists no negative tissue because AXIN2 is detected in all 45 scored tissues, so use no-primary and isotype controls; compare adjacent non-glandular cells as an internal background reference without assuming they are AXIN2-negative (HPA: no negative tissue listed).
Positive control tissue: Breast (Adipocytes, HPA High)
Negative control tissue: None in HPA: AXIN2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AXIN2 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (uncertain), Plasma membrane (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a host- and clonality-matched isotype control, alongside AXIN2-knockout tissue or a validated immunizing-peptide block as a biological specificity control (A01772-2 tissue-IHC caption: rabbit primary). For chromogenic colon IHC, quench endogenous peroxidase and check background in the no-primary slide, especially around blood or inflammatory cells (standard IHC practice).
⚠️Feasibility: The A01772-2 paraffin-section example uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for every antibody or specimen (A01772-2 tissue-IHC caption). Its fixative is unreported, and the supplied evidence reports no AXIN2-specific fixation window or fixation effect (A01772-2 tissue-IHC caption). There is no basis here to rank frozen sections or IF as easier; in colon, inspect chromogenic background from endogenous peroxidase before scoring glandular staining (standard IHC practice).
HPA tissue IHC evidence for AXIN2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: AXIN2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced AXIN2 IHC Tips
Troubleshoot AXIN2 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting signal as Wnt pathway activity.
What should I change when AXIN2 staining is weak after antigen retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A01772-2). The catalog antibody detected AXIN2 in a paraffin section after this treatment, followed by 2 μg/ml primary antibody overnight at 4°C (A01772-2 caption). If staining is weak, check heating consistency and section adhesion before testing another retrieval condition on adjacent sections (standard IHC practice). Compare each condition with the same detection reagents and a positive tissue control, such as colon glandular cells, which show high staining in the reference profile (HPA: High in colon glandular cells). Avoid interpreting stronger background alone as improved retrieval (standard IHC practice).
Can fixation explain weak or patchy AXIN2 staining?
Target-specific AXIN2 sensitivity to fixation is unknown from the supplied evidence (A01772-2 caption: fixative not stated). The catalog image shows staining in a paraffin-embedded human renal carcinoma section, but its caption does not identify the fixative (A01772-2 caption). Record the specimen's actual fixation and processing history, then compare sections processed together before changing antibody conditions (standard IHC practice). Patchy staining that follows folds, damaged areas or section edges should prompt a processing check and review of an adjacent stained section (standard IHC practice). Do not attribute such variation to an AXIN2-specific fixation effect without a controlled comparison (standard IHC practice).
How should I judge nuclear or membrane-like AXIN2 staining in IHC?
Use cytoplasmic staining as the strongest compartmental expectation: UniProt places AXIN2 in the cytoplasm, and HPA supports cytosolic localisation (UniProt Q9Y2T1: subcellular location; HPA: cytosol supported). HPA tissue IHC also describes general cytoplasmic and nuclear expression, so nuclear signal warrants evaluation alongside controls (HPA: tissue IHC profile). HPA labels nucleoplasmic and plasma-membrane localisation uncertain, while the supplied sequence record has no transmembrane segment (HPA: subcellular; UniProt Q9Y2T1: topology). For a membrane-like rim, inspect cell boundaries and adjacent sections; confirm that the signal persists with appropriate antibody and detection controls before assigning it to AXIN2 (standard IHC practice).
Could an AXIN2 isoform or hidden epitope explain inconsistent staining?
The supplied record lists 0 isoforms and does not specify this antibody's epitope, so an isoform-specific explanation cannot be established here (UniProt Q9Y2T1: isoforms; A01772-2 caption). AXIN2 has an RGS domain at residues 81–200 and a DIX domain at 761–843, but neither location identifies the antibody-binding site (UniProt Q9Y2T1: domains). The record lists no glycosylation sites or mapped modified residues; that annotation does not establish how fixation affects epitope access (UniProt Q9Y2T1: annotations). Compare retrieval conditions on matched sections and seek an independently validated antibody with a documented epitope if epitope masking remains suspected (standard IHC practice).
How can IF help check an ambiguous chromogenic AXIN2 pattern?
On an adjacent section, multiplex AXIN2 with a marker identifying the cell population being assessed; HPA reports high AXIN2 staining in colon glandular cells and lung macrophages (HPA: tissue IHC profile). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before calling a faint signal positive (standard IF practice). Because supported AXIN2 localisation is cytosolic, access to an intracellular epitope generally requires permeabilisation; a surface-only interpretation is weaker because plasma-membrane localisation is uncertain and AXIN2 has no transmembrane segment (HPA: subcellular; UniProt Q9Y2T1: topology; standard IF practice). Keep the IF result as a compartment check alongside the page's chromogenic IHC evidence (A01772-2 caption; standard IHC practice).
What causes diffuse AXIN2 background in DAB-stained paraffin sections?
First compare the pattern with a no-primary control and inspect whether colour follows tissue edges, folds or damaged areas (standard IHC practice). The documented chromogenic workflow used 10% goat serum blocking, 2 μg/ml rabbit primary antibody overnight at 4°C, a peroxidase-conjugated secondary and DAB (A01772-2 caption). If the no-primary section also develops colour, review endogenous peroxidase blocking and secondary-antibody binding as general workflow causes (standard IHC practice). If background appears only with primary antibody, compare matched sections after adjusting blocking and washing, while retaining a positive tissue control (standard IHC practice). Score cellular signal separately from extracellular deposits (standard IHC practice).
How should I quantify AXIN2 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; cytoplasmic signal has supported localisation, while nuclear staining appears in the HPA tissue profile (HPA: subcellular; HPA: tissue IHC profile). For a defined population, record the percentage of positive cells and an intensity-based H-score from 0–300, using the same scoring rules across sections (standard IHC practice). Normalise cell counts to the number of evaluable cells, or report positive-cell density per mm² of viable tissue when area matters (standard IHC practice). Exclude necrosis, folds and poorly preserved edges consistently, and record cytoplasmic and nuclear scores separately if both are measured (standard IHC practice).
When is AXIN2 staining convincing rather than artefactual?
A convincing result shows reproducible cellular staining in the expected compartment and a plausible cell population, with clean detection controls (UniProt Q9Y2T1: cytoplasm; HPA: tissue IHC profile; standard IHC practice). High staining in colon glandular cells or lung macrophages can inform positive-control selection, but those patterns alone do not establish antibody specificity in a new specimen (HPA: tissue IHC profile; standard IHC practice). Treat isolated membrane rims cautiously because membrane localisation is uncertain and AXIN2 lacks a transmembrane segment (HPA: subcellular; UniProt Q9Y2T1: topology). Discount signal confined to edges, necrosis or a no-primary control, including colour from endogenous peroxidase (standard IHC practice).
Boster reagents
Best AXIN2 / Axin-2 IHC Antibodies
The catalog antibodies have IHC images from human renal carcinoma, spleen and thyroid cancer sections (catalog images: A01772-2), and rat and mouse lung tissue, plus IF images from rat lung cells, mouse lung tissue and HepG2 cells (catalog images: A01772).
Real IHC data
Anti-Axin 2/AXIN2 Antibody ®
Cat # A01772-2
Real IHC data
Anti-Axin-2 AXIN2 Antibody
Cat # A01772
A01772-2 has IHC images from paraffin-embedded human renal carcinoma, spleen and thyroid cancer sections (catalog images: A01772-2). A01772 has IHC images from rat and mouse lung tissue and IF images from rat lung cells, mouse lung tissue and HepG2 cells (catalog images: A01772).
Which to pick: For human paraffin-section IHC, choose A01772-2: its caption documents EDTA retrieval at pH 8.0 and staining at 2 μg/ml; the fixative is unreported (catalog IHC caption: A01772-2). For IF/ICC work, choose A01772 for its IF images and 20 μg/mL starting concentration; ICC validation is unreported (catalog IF images and datasheet: A01772). For mouse or rat tissue IHC, choose A01772: its IHC images show lung tissue from both species at 5 μg/mL; its listed reactivity also includes human (catalog IHC images and reactivity: A01772).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y2T1 (AXIN2_HUMAN, Axin-2).
- Human Protein Atlas. AXIN2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. AXIN2 subcellular location (ICC-IF): Localized to the nucleoplasm, plasma membrane and cytosol..
- Human Protein Atlas. AXIN2 antibody validation summary (2 antibodies).
- Activation of the Wnt/β-catenin signaling pathway by mechanical ventilation is associated with ventilator-induced pulmonary fibrosis in healthy lungs. PloS one 2011 — PMC3174135.
- Expression of β-catenin and AXIN2 in ameloblastomas. Contemporary oncology (Poznan, Poland) 2013 — PMC3934066.
- Niclosamide and Pyrvinium Are Both Potential Therapeutics for Osteosarcoma, Inhibiting Wnt-Axin2-Snail Cascade. Cancers 2021 — PMC8464802.
- Beneficial effects of an endogenous enrichment in n3-PUFAs on Wnt signaling are associated with attenuation of alcohol-mediated liver disease in mice. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2021 — PMC8243414.
- PubMed PMID:10049590 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AXIN2 (Axin-2). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01772-2). Validated IHC antibody: A01772-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human renal carcinoma tissue; fixative not specified (datasheet A01772-2); 2. Fixation: Image fixative and duration unreported (datasheet A01772-2); 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 A01772-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01772-2); 8. Primary antibody: Rabbit anti-AXIN2, 2-5 μg/ml (datasheet A01772-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01772-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A01772-2); verify before use.; Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 0 listed isoforms; no processing reported (UniProt); Regulation: Intensity regulation unreported (UniProt). Evidence: UniProt Q9Y2T1, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm, plasma membrane and cytosol., Datasheet A01772-2, PMC3174135, PMC3934066. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AXIN2/immunohistochemistry-ihc-guide.