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DACT2 / Dapper homolog 2 · IHC design guide
Design Immunohistochemistry for DACT2
Plan chromogenic DACT2 IHC-P around its broad cytoplasmic tissue staining (HPA tissue IHC). Use hepatocytes, syncytiotrophoblasts or prostate glandular cells as positive tissue references (HPA tissue IHC), and start the catalog antibody at 2.5 μg/mL (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DACT2 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 many tissues (HPA tissue IHC) | |
| Staining pattern | Broad cytoplasmic staining; strong in hepatocytes (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Liver+4 more · see all | |
| Negative control | Parathyroid gland+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium agreement with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | Four isoforms; assess antibody epitope coverage (UniProt) |
Section 1
Recommended DACT2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published chromogenic DACT2 protocols from HCC, myocardial, and esophageal carcinoma specimens (PMC3605395; PMC7330378; PMC5225534).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A10097); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-DACT2, 2.5 μg/mL (datasheet A10097) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DACT2-positive staining in hepatocytes of liver (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval guidance); adjust against the published myocardial protocol (PMC7330378).
Section 2
What Is the Expected DACT2 Staining Pattern?
Expect predominantly cytoplasmic DACT2 staining in paraffin sections, with strong signal in hepatocytes, placental syncytiotrophoblasts and prostate glandular cells (HPA: ubiquitous cytoplasmic profile; High in these cells). DACT2 has no annotated transmembrane segment (UniProt Q5SW24 topology). HPA rates its IHC pattern Approved, with medium consistency against RNA data and external verification pending (HPA: reliability).
| Clear cytoplasmic signal in hepatocytes, syncytiotrophoblasts or prostate glandular cells. | This fits the reported compartment and high-staining cell populations (HPA: cytoplasmic profile; High in liver, placenta and prostate). Judge signal in the named cells, alongside a negative control, rather than treating every stained structure in the section as DACT2 (general IHC practice). |
| Predominantly crisp membrane or nuclear staining, with little cytoplasmic signal. | This conflicts with the reported cytoplasmic IHC profile (HPA: tissue IHC profile). A dominant membrane rim is also unsupported by the absence of an annotated transmembrane segment (UniProt Q5SW24 topology). Consider localization artefact or nonspecific staining; check controls before interpreting a new compartment. |
| Strong signal in cells reported as unstained, especially parathyroid or thyroid glandular cells. | HPA reports DACT2 as not detected in those glandular cells (HPA: parathyroid and thyroid gland). Cross-reactivity or endogenous detection activity is possible (general IHC practice). Compare the staining with a no-primary control and with an HPA high-staining cell population. |
| Diffuse color across cells and surrounding tissue, without discernible cytoplasmic boundaries. | A widespread haze cannot establish the reported cellular pattern (HPA: cytoplasmic profile). Background from detection reagents or inadequate blocking can obscure a specific signal (general IHC practice). Use the negative control to identify background before scoring cell-specific staining. |
| No signal in hepatocytes, placental syncytiotrophoblasts or prostate glandular cells. | Each is reported as a high-staining population (HPA: tissue IHC). Absence in an otherwise suitable positive-control section suggests an assay or specimen problem (general IHC practice). HPA's Approved rating has medium RNA agreement and awaits external verification, so a single negative result does not settle expression (HPA: reliability). |
💡Expected DACT2 appearanceA convincing positive is cell-resolved, predominantly cytoplasmic staining, strongest in HPA high-staining hepatocytes, syncytiotrophoblasts or prostate glandular cells; isolated nuclear or membrane staining, or color in HPA unstained glandular cells, needs control-based review (HPA: tissue IHC profile and staining levels; UniProt Q5SW24 topology).
| Tissue and cell choice | HPA reports High staining in hepatocytes, syncytiotrophoblasts and prostate glandular cells; it reports Not detected in parathyroid and thyroid glandular cells (HPA: tissue IHC). Those named populations provide more interpretable comparisons than a whole-section positive or negative label. |
| Strength of localization evidence | The cytoplasmic pattern comes from HPA tissue IHC, rated Approved with medium RNA consistency and pending external verification (HPA: profile and reliability). UniProt supplies no subcellular annotation, and HPA lists no ICC-IF images or main IF location (UniProt Q5SW24; HPA: subcellular). An IF result therefore needs its own controls. |
| Topology and processing | DACT2 has no annotated transmembrane segment or signal peptide, and UniProt lists one chain spanning residues 1–774 (UniProt Q5SW24 topology and processing). These annotations support checking a dominant membrane-only pattern; they do not identify the antibody epitope or establish a retrieval condition. |
| Isoform coverage | UniProt lists four DACT2 isoforms (UniProt Q5SW24 isoforms). The supplied record gives no antibody epitope, so coverage across isoforms cannot be inferred. If staining differs between samples, keep that uncertainty in mind before attributing the difference to expression. |
| Detection activity | For a peroxidase-based chromogenic assay, endogenous activity can create color independent of primary-antibody binding (general IHC practice). A no-primary control and an appropriate enzyme-blocking step help distinguish that background from the cell-resolved HPA pattern (HPA: cytoplasmic profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining cells are blank. | The assay may have failed, or the section may not contain an interpretable positive population (general IHC practice; HPA: High in hepatocytes, syncytiotrophoblasts and prostate glandular cells). | Confirm the named cells are present, inspect the counterstain and detection controls, then review retrieval and antibody incubation using the assay's validated instructions (general IHC practice). No DACT2-specific retrieval setting is supplied. |
| The entire section develops color, including a no-primary control. | Primary-antibody binding cannot explain color in that control; endogenous enzyme activity or detection-reagent background is possible (general IHC practice). | Check the detection system's blocking and wash steps, and compare control and test sections before scoring cytoplasmic staining (general IHC practice; HPA: cytoplasmic profile). |
| Parathyroid or thyroid glandular cells stain strongly. | That finding conflicts with HPA's Not detected calls for these cells; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Compare a no-primary control and an HPA high-staining population on the same run; interpret persistent unexpected staining cautiously (general IHC practice; HPA: tissue IHC). |
| Staining appears mainly as a membrane rim or nuclear focus. | This differs from the HPA cytoplasmic profile; a membrane-only result also lacks support from UniProt topology (HPA: tissue IHC profile; UniProt Q5SW24 topology). | Inspect cell boundaries and the negative control, then check whether detection background or section artefact explains the pattern before assigning DACT2 localization (general IHC practice). |
| Signal is weak or uneven in a reported high-staining population. | Uneven section quality or assay conditions can complicate comparison (general IHC practice); HPA's staining levels do not establish a DACT2-specific fixation effect (HPA: tissue IHC). | Check section integrity, retrieval and incubation against the validated IHC instructions, and compare a concurrently processed positive control (general IHC practice). |
| An IF image appears to show a precise DACT2 compartment. | The supplied HPA subcellular record has no main location or ICC-IF images; tissue IHC alone cannot validate that IF assignment (HPA: subcellular and tissue IHC). | Assess the IF signal with modality-appropriate negative controls and cell markers before reporting a compartment (general IF practice); use the separate IF/ICC guide for assay design. |
Experiment evidence
IHC & IF images showing DACT2 staining
A10097 · IHCImmunohistochemistry of Dact2 in human brain tissue with Dact2 antibody at 2.5 μg/mL.
A10097 · IFImmunofluorescence of Dact2 in Human Brain cells with Dact2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164488-DACT2/tissue/cerebral+cortex#imid_7605632
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164488-DACT2/tissue/colon#imid_7605558
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164488-DACT2/tissue/liver#imid_7605551
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164488-DACT2/tissue/kidney#imid_7605552
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DACT2 IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: High in hepatocytes). Use parathyroid gland as the negative tissue (HPA: Not detected in glandular cells); on the liver slide, cells without specific staining should remain at background, but no particular non-hepatocyte cell type is established as DACT2-negative by the supplied evidence (HPA: liver row specifies hepatocytes only).
Positive control tissue: Liver (Hepatocytes, HPA High)
Negative control tissue: Parathyroid gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for DACT2; derive a cell-line control from the positive tissue's cell type (Hepatocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched isotype control of the same host species and clonality; and a DACT2 knockout specimen or validated peptide-block control (standard IHC practice). For liver, quench endogenous peroxidase and check for endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A DACT2-specific fixation window and retrieval effect are unreported in the supplied evidence; assess antigen retrieval empirically for paraffin sections (standard IHC practice). The selected A10097 tissue-IHC caption does not report a fixative (selected tissue-IHC caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier for DACT2; liver pigment may complicate chromogen interpretation, so compare with the no-primary slide (standard IHC practice).
HPA tissue IHC evidence for DACT2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Placenta | Syncytiotrophoblasts | High | Protein (IHC) | HPA → |
| Prostate | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced DACT2 IHC Tips
Troubleshoot DACT2 staining in paraffin sections by checking retrieval, cell type, cytoplasmic localisation, and controls before interpreting signal intensity.
How should I retrieve DACT2 in paraffin sections when cytoplasmic staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval rule). Let slides cool in the retrieval solution, then compare the same tissue and antibody concentration across runs so changes in staining can be attributed to retrieval (standard IHC practice). If staining remains weak, test a longer retrieval interval or an alternative buffer on adjacent sections, while checking whether tissue structure and background deteriorate (standard IHC practice). Score the expected cytoplasmic signal in the same cell population rather than treating any darker precipitate as improvement (HPA: ubiquitous cytoplasmic expression; standard IHC practice).
Could fixation explain weak or patchy DACT2 staining?
DACT2-specific sensitivity to fixation is unknown because the supplied tissue image caption does not state a fixative (A10097 tissue-IHC caption). Record the fixative, fixation duration, tissue thickness, and processing history for each block before comparing staining between specimens (standard IHC practice). If processing varies, stain adjacent sections from a consistently processed control block alongside the test sections and apply the same citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule; standard IHC practice). Interpret any change as a processing-associated observation until a controlled comparison establishes whether fixation itself affected DACT2 detection (standard IHC practice).
Where should DACT2 staining appear, and when is a different compartment suspicious?
Assess cytoplasmic staining first because the tissue IHC profile describes ubiquitous cytoplasmic expression, while UniProt supplies no subcellular location for DACT2 (HPA: ubiquitous cytoplasmic expression; UniProt Q5SW24: location not annotated). DACT2 has no annotated transmembrane segment, so a crisp membrane-only outline needs independent validation before assignment to this target (UniProt Q5SW24 topology; standard IHC practice). Compare high-signal hepatocytes or syncytiotrophoblasts with low-signal squamous cells of oral mucosa on matched runs where available (HPA: high in hepatocytes and syncytiotrophoblasts; low in oral mucosa). Keep counterstaining consistent and inspect cytoplasm at the same magnification before scoring a shift toward nuclear or membrane signal (standard IHC practice).
How can DACT2 isoforms or an unknown epitope complicate IHC interpretation?
DACT2 has 4 annotated isoforms, but the supplied material does not map the catalog antibody’s epitope or establish which isoforms it detects (UniProt Q5SW24: isoforms 1–4; supplied antibody evidence). Treat staining as antibody-reactive DACT2 signal rather than an isoform-specific measurement until epitope mapping or an orthogonal assay supports that assignment (standard IHC practice). Compare adjacent sections with and without citrate pH 6.0 retrieval for 20 min, keeping detection conditions fixed, to identify retrieval-dependent changes without assigning them to an isoform (page retrieval rule; standard IHC practice). If results differ across specimens, document the antibody identity, retrieval conditions, and cell-level pattern before proposing a splice-related explanation (standard IHC practice).
How should I check DACT2 by multiplex IF alongside chromogenic IHC?
For a separate IF assay, pair DACT2 with a validated marker for the cell population being examined, such as hepatocytes or syncytiotrophoblasts, and compare cell-level overlap with the IHC pattern (HPA: high in hepatocytes and syncytiotrophoblasts; standard IF practice). Include single-stain and no-primary controls, and place the weaker signal in a far-red channel if tissue autofluorescence is prominent (standard IF practice). Because tissue IHC describes cytoplasmic DACT2 and UniProt lists no transmembrane segment, test permeabilisation when the antibody epitope is intracellular; its exact epitope is not supplied (HPA: ubiquitous cytoplasmic expression; UniProt Q5SW24 topology; supplied antibody evidence). Optimise IF fixation and permeabilisation independently, since the tissue-IHC caption gives neither IF conditions nor fixation details (A10097 tissue-IHC caption; standard IF practice).
What controls help distinguish DACT2 signal from chromogenic background?
Run a no-primary control and inspect nonspecific precipitate, endogenous peroxidase activity, and pigment before changing the DACT2 antibody concentration (standard IHC practice). Apply a peroxidase block before chromogenic detection and use matched blocking and washing conditions across sections; these are general workflow steps, not DACT2-specific validation (standard IHC practice). The selected brain tissue caption reports 2.5 µg/mL antibody but gives no fixative or background-control results, so use that concentration as a documented reference point rather than a guaranteed optimum (A10097 tissue-IHC caption). Compare cytoplasmic staining with the no-primary control and review section edges separately, where uneven processing can confound interpretation (HPA: cytoplasmic expression; standard IHC practice).
How should I quantify DACT2 across sections with different cellular composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, then record cytoplasmic intensity and the percentage of positive cells using identical imaging and detection settings (HPA: cytoplasmic expression; standard IHC practice). An H-score sums the percentage of cells at each intensity grade multiplied by that grade, giving a 0–300 range when grades are 0–3 (standard IHC practice). Report the H-score or positive-cell percentage per relevant cell population, and use positive-cell density per mm² only when tissue area is the intended denominator (standard IHC practice). Normalise comparisons to the same cell type and viable tissue area, with a consistently processed control section in each run (standard IHC practice).
When is apparent DACT2 positivity more likely to be artefact?
Give greatest weight to reproducible cytoplasmic staining in the expected cells: hepatocytes and syncytiotrophoblasts are reported high, whereas thyroid and parathyroid glandular cells are reported undetected (HPA: tissue IHC profile). A membrane-only or predominantly nuclear pattern warrants checking antibody specificity because the tissue profile is cytoplasmic and UniProt provides no subcellular annotation (HPA: ubiquitous cytoplasmic expression; UniProt Q5SW24: location not annotated). Exclude section-edge staining, necrotic regions, pigment, and signal retained in the no-primary control before calling cells positive (standard IHC practice). Treat agreement with the HPA pattern as supportive rather than conclusive because its IHC reliability is approved with medium RNA–staining consistency and pending external verification (HPA: reliability description).
Boster reagents
Best DACT2 / Dapper homolog 2 IHC Antibodies
A10097 has IHC data from human brain tissue (IHC image caption) and IF data from human brain cells (IF image caption).
Real IHC data
Anti-Dapper homolog 2 Dact2 Antibody
Cat # A10097
A10097 is listed for IHC-P and shown staining human brain tissue at 2.5 μg/mL (catalog applications; IHC image caption). A10097 is also listed for IF and shown staining human brain cells at 20 μg/mL (catalog applications; IF image caption).
Which to pick: Choose A10097 for paraffin-section IHC (catalog applications: IHC-P); its IHC image shows human brain tissue, but the fixative is unreported (IHC image caption). For IF/ICC, A10097 has IF data from human brain cells (IF image caption); ICC validation is unreported (catalog applications). For mouse or rat samples, A10097 lists reactivity with both species (catalog reactivity), while its IHC and IF images show human samples (IHC and IF image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5SW24 (DACT2_HUMAN, Dapper homolog 2).
- Human Protein Atlas. DACT2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DACT2 subcellular location (ICC-IF): Highest expression in OE19: 18.8 nTPM.
- Human Protein Atlas. DACT2 antibody validation summary (1 antibodies).
- Reduced expression of DACT2 promotes hepatocellular carcinoma progression: involvement of methylation-mediated gene silencing. World journal of surgical oncology 2013 — PMC3605395.
- DACT2 regulates structural and electrical atrial remodeling in atrial fibrillation. Journal of thoracic disease 2020 — PMC7330378.
- Dact2 represses PITX2 transcriptional activation and cell proliferation through Wnt/beta-catenin signaling during odontogenesis. PloS one 2013 — PMC3551926.
- Aberrant methylation of DACT1 and DACT2 are associated with tumor progression and poor prognosis in esophageal squamous cell carcinoma. Journal of biomedical science 2017 — PMC5225534.
- PubMed PMID:15498874 — UniProt-cited evidence.
- PubMed PMID:14574404 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DACT2 (Dapper homolog 2). Expected localisation Cytoplasmic staining in many tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10097 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Parathyroid gland. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A10097); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-DACT2, 2.5 μg/mL (datasheet A10097); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has medium agreement with RNA data (HPA tissue IHC); Isoform / epitope: Four isoforms; assess antibody epitope coverage (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q5SW24, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in OE19: 18.8 nTPM, Datasheet A10097, PMC3605395, PMC7330378. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DACT2/immunohistochemistry-ihc-guide.