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DAB1 / Disabled homolog 1 · IHC design guide
Design Immunohistochemistry for DAB1
Plan DAB1 paraffin-section IHC using intestinal enterocyte microvilli and endocrine cells as positive staining references (HPA tissue IHC). The catalog antibody lists an IHC dilution range of 1:50–1:200 (datasheet).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DAB1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic and membranous in small intestine (HPA tissue IHC) | |
| Staining pattern | Enterocyte microvilli and intestinal endocrine cells stain (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Duodenum+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 paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Brain expression may not predict intestinal IHC staining (UniProt; HPA tissue IHC) | |
| Regulation | Expression regulation unspecified (UniProt) | |
| Isoform / epitope | Six isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended DAB1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with published DAB1 IHC methods for liver (PMC5365015) and human brain (PMC12191094).
| Sample | Paraffin-embedded human small intestine tissue; fixative not specified (datasheet A03459) |
| Fixation | Image fixative and duration unreported (datasheet A03459); 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-DAB1, 1:50-1:200 (datasheet A03459) |
| 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 | DAB1-positive staining in enterocytes - Microvilli of duodenum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in small intestines. Low expression in CNS and retina. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); citrate pH 6 was also used in liver (PMC5365015).
Section 2
What Is the Expected DAB1 Staining Pattern?
DAB1 is a cytoplasmic protein with no transmembrane segment (UniProt O75553 localization and topology). In paraffin-section IHC, expect cytoplasmic and membranous staining in small-intestinal cells, especially duodenal enterocyte microvilli and small-intestinal endocrine cells (HPA: High; tissue IHC reliability Enhanced). Neuronal staining can be weaker or more selective: HPA reports Medium staining in cerebral-cortex neurons, cerebellar molecular-layer cells and retinal ganglion cells, alongside an overall low CNS and retinal profile (HPA tissue IHC).
| Strong microvillar staining in duodenal enterocytes, with cytoplasmic or membranous staining in small-intestinal endocrine cells. | This fits the most prominent reported IHC pattern (HPA: High in duodenal enterocyte microvilli and small-intestinal endocrine cells). Score the named cells and compartment; an intense signal elsewhere in the section does not establish the same DAB1 pattern (HPA tissue IHC). |
| Moderate staining in cerebral-cortex neurons, cerebellar molecular-layer cells or retinal ganglion cells. | These are reported positive cell populations (HPA: Medium in each). HPA also describes overall low CNS and retinal expression, so compare the specified cells rather than expecting uniformly strong staining throughout either tissue (HPA tissue IHC). |
| Predominantly nuclear staining, with little staining in the expected cytoplasmic or small-intestinal membranous sites. | A nuclear-dominant result conflicts with the reported localization and IHC pattern (UniProt O75553: Cytoplasm; HPA tissue IHC: cytoplasmic and membranous in small intestine). Treat it as suspect and assess morphology and staining controls before assigning target-specific signal (general IHC practice). |
| Strong staining in an HPA-listed undetected cell population, such as adipocytes or bronchial respiratory epithelial cells. | This disagrees with those reference observations (HPA: Not detected in adipocytes of adipose tissue and respiratory epithelial cells of bronchus). Cross-reactivity or endogenous detection activity are possibilities, not diagnoses; compare controls and the staining pattern in a known-positive section (general IHC practice). |
| Diffuse color across cells and extracellular areas, or no stain in a known-positive small-intestinal cell population. | Diffuse noncellular color limits localization scoring; absent signal in a positive comparator makes a negative test section uninterpretable (general IHC practice). The reference comparators are duodenal enterocyte microvilli and small-intestinal endocrine cells (HPA: High); neither outcome alone proves altered DAB1 expression. |
💡Expected DAB1 appearanceCall a convincing positive when duodenal enterocyte microvilli or small-intestinal endocrine cells show strong, cell-associated cytoplasmic or membranous signal (HPA: High; HPA tissue IHC profile); nuclear-dominant or diffuse noncellular color is suspect against the reported pattern (UniProt O75553: Cytoplasm; general IHC practice).
| Cell population and region | The reference is cell-specific: duodenal enterocyte microvilli and small-intestinal endocrine cells are High, while specified cortical, cerebellar and retinal cells are Medium (HPA tissue IHC). A whole-tissue average can obscure those differences (general IHC scoring practice). |
| Topology and processing | DAB1 lacks a transmembrane segment and signal peptide, and UniProt lists one chain spanning residues 1–588 (UniProt O75553 topology and processing). Interpret an apical microvillar IHC signal as the observed cellular pattern, without inferring that DAB1 is an integral membrane protein (HPA tissue IHC; UniProt O75553 topology). |
| Isoforms and antibody recognition | UniProt lists six DAB1 isoforms, but the payload supplies no catalog-antibody epitope or isoform coverage (UniProt O75553 isoforms; supplied antibody data). A change in staining cannot be assigned to one isoform from this evidence; use antibody-specific documentation before making that claim (general IHC interpretation practice). |
| Antibody validation | HPA marks IHC for HPA052033 and CAB032329 as Enhanced; HPA067495 has no listed IHC status (HPA antibody validation). This supports using the validated IHC observations as comparators, while leaving performance of an unspecified catalog antibody to be checked on the actual sections (general IHC practice). |
| IF/ICC Q: Should vesicles define the paraffin-IHC call? | A: HPA reports enhanced vesicular localization in ICC-IF images from HeLa and RT-4 cells (HPA subcellular ICC-IF). For paraffin IHC, use the cell and compartment pattern reported in tissue sections; the ICC-IF observation does not establish an IHC protocol or replace that tissue reference (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in duodenal enterocyte microvilli or small-intestinal endocrine cells. | These are High reference populations (HPA tissue IHC); a failed run or unsuitable antibody conditions are possibilities (general IHC practice). | Check section integrity, positive-control staining, antibody conditions and chromogenic detection. If the positive control fails, repeat the run before scoring the test section negative (general IHC practice). |
| Signal is mainly nuclear. | That distribution conflicts with cytoplasmic localization and the reported intestinal pattern (UniProt O75553; HPA tissue IHC). | Review morphology and negative controls, then compare a known-positive intestinal section; do not score nuclear-only color as the expected DAB1 pattern (general IHC practice). |
| Widespread color obscures cell borders or persists outside cells. | Nonspecific background or endogenous detection activity may contribute (general chromogenic IHC practice). | Inspect negative controls and reagent-only controls where appropriate; check blocking, washing and detection steps, then judge only localized cellular staining (general IHC practice). |
| Strong color appears in HPA-listed undetected cells. | Reference examples include adipose-tissue adipocytes and bronchial respiratory epithelial cells (HPA: Not detected); cross-reactivity or detection background may explain discordance (general IHC practice). | Compare the same run with a known-positive intestinal section and relevant negative controls; report persistent discordance as unresolved rather than calling it DAB1 expression (general IHC practice). |
| Brain or retinal sections look weak beside small intestine. | HPA reports Medium staining in specified cortical, cerebellar and retinal cells, and describes overall low CNS and retinal expression (HPA tissue IHC). | Score the named cell populations separately and compare them with a positive intestinal control; do not require brain or retina to match the High intestinal intensity (HPA tissue IHC; general IHC scoring practice). |
| Only a subset of cells stains within an apparently positive tissue. | The HPA observations name particular cell populations and compartments; the data do not establish uniform staining across a whole tissue (HPA tissue IHC). | Annotate the stained cell type and compartment, then compare those observations with the relevant HPA entry before interpreting unstained neighboring cells (general IHC scoring practice). |
Experiment evidence
IHC & IF images showing DAB1 staining
A03459 · IHCImmunohistochemistry (IHC) analyzes of Dab1 (Q226) pAb in paraffin-embedded human small intestine tissue at 1:100.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000173406-DAB1/tissue/cerebral+cortex#imid_17239315
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000173406-DAB1/tissue/small+intestine#imid_17239200
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000173406-DAB1/tissue/colon#imid_17239233
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000173406-DAB1/tissue/liver#imid_17239234
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052033; green = DAB1, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052033; green = DAB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DAB1 IHC & IF
🧪Run duodenum first: enterocyte microvilli should stain (High; HPA: duodenum, enterocytes–microvilli). Use adipose tissue adipocytes as the negative tissue (HPA: adipocytes not detected); on the duodenum slide, cells outside the expected microvillar staining pattern should show little signal, but they are background comparators rather than validated DAB1-negative cells (HPA: duodenum, enterocytes–microvilli).
Positive control tissue: Duodenum (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DAB1 in HeLa, RT-4, with annotated localisation: Vesicles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, host- and isotype-matched nonimmune immunoglobulin for the polyclonal catalog antibody, and DAB1-knockout tissue if available as a biological specificity control (caption: pAb; standard IHC practice). Block endogenous peroxidase for chromogenic detection and check the duodenal luminal edge for background that could mimic microvillar staining (HPA: duodenum, enterocytes–microvilli; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected A03459 paraffin-section caption does not state the fixative (caption: paraffin-embedded human small intestine at 1:100; fixative unreported). Antigen-retrieval dependence is unreported, so assess retrieval conditions empirically; the supplied evidence does not establish whether frozen sections or IF are easier (standard IHC practice; supplied HPA and caption evidence). In duodenum, distinguish staining at enterocyte microvilli from luminal-edge background with the controls above (HPA: duodenum, enterocytes–microvilli; standard IHC practice).
HPA tissue IHC evidence for DAB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Duodenum | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Small intestine | Endocrine cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Retina | Ganglion cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced DAB1 IHC Tips
Troubleshoot DAB1 staining in paraffin sections by checking retrieval, tissue context, compartment, and controls before scoring chromogenic signal.
What retrieval conditions should I try first for DAB1 in paraffin sections?
Use citrate buffer at pH 6.0 for heat induced antigen retrieval at 95–98 °C for 20 min (this page’s retrieval rule). Let sections cool in buffer, then rinse consistently before blocking so staining can be compared across runs (standard IHC practice). If staining is weak, vary retrieval time on adjacent sections while keeping antibody concentration and detection conditions fixed (standard IHC practice). Include a small intestine section with the test sections, since the selected catalog antibody has a paraffin section image from human small intestine at 1:100 (A03459 tissue IHC caption). Do not treat the caption as evidence for a particular fixative or retrieval method (A03459 tissue IHC caption).
How should I troubleshoot weak DAB1 staining when fixation may vary?
The supplied evidence does not establish DAB1 specific sensitivity to any fixative or fixation duration (A03459 tissue IHC caption: fixative unstated). Record the fixative, time before fixation, and fixation duration for each specimen, then compare sections processed and stained together (standard IHC practice). If fixation history differs, use a consistent processing series before changing retrieval or antibody concentration, because several preparation variables can affect staining (standard IHC practice). The selected antibody’s human small intestine image documents paraffin sections at 1:100, without reporting fixation conditions (A03459 tissue IHC caption). Interpret a weak result against matched controls rather than attributing it to DAB1 biology (standard IHC practice).
Which staining compartments fit DAB1, and how should membranous signal be judged?
Expect intracellular staining because DAB1 is annotated as cytoplasmic and has no transmembrane segment (UniProt O75553 subcellular annotation; UniProt O75553 topology). Tissue IHC also reports cytoplasmic and membranous expression in small intestines, including high staining of duodenal enterocyte microvilli (HPA tissue IHC profile; HPA: High in duodenal enterocyte microvilli). Assess apparent membrane accentuation alongside cytoplasmic staining and the relevant cell type, rather than treating every outlined cell as positive (HPA tissue IHC profile; standard IHC practice). DAB1 is also reported at vesicles in cell imaging, which may help frame punctate signal but does not establish its appearance in chromogenic tissue sections (HPA subcellular: vesicles; standard IHC practice). Compare the pattern with a matched negative control (standard IHC practice).
Could splice variants or phosphorylation explain inconsistent DAB1 IHC staining?
DAB1 has 6 listed isoforms, so an antibody’s recognized sequence matters when comparing tissue patterns (UniProt O75553 isoforms). Its PID domain spans residues 36–189, and reported modifications include phosphotyrosines at 198, 220, and 232 plus CDK5 associated phosphoserine at 524 (UniProt O75553 domains; UniProt O75553 modified residues). The supplied caption does not identify the selected antibody’s epitope or show which isoforms it recognizes (A03459 tissue IHC caption). Check epitope documentation before assigning discordant staining to a splice variant or phosphorylation state (standard IHC interpretation). Compare sections with the same processing and detection conditions while resolving that uncertainty (standard IHC practice).
How can I use IF to investigate an ambiguous DAB1 IHC pattern?
Use IF as a separate localization check: DAB1 is annotated as cytoplasmic, while cell imaging reports vesicles (UniProt O75553 subcellular annotation; HPA subcellular: vesicles). Multiplex with a validated marker for the expected cell population, such as the neuronal cells reported in cerebral cortex or the endocrine cells reported in small intestine, and assess overlap at the cell level (HPA: Medium in cerebral cortex neuronal cells; HPA: High in small intestine endocrine cells; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence and appropriate single stain controls (standard IF practice). Because DAB1 lacks a transmembrane segment, use permeabilisation suitable for an intracellular epitope, guided by the IF antibody’s epitope information (UniProt O75553 topology; standard IF practice).
What should I check when DAB1 DAB staining is widespread?
Run a no primary control and inspect whether the same deposits persist, particularly where endogenous peroxidase or nonspecific detection could contribute signal (standard chromogenic IHC practice). Confirm peroxidase blocking, protein blocking, wash consistency, and DAB development time on matched sections before adjusting the primary antibody (standard chromogenic IHC practice). The selected image reports 1:100 in human small intestine paraffin sections, but does not establish an optimal dilution for other specimens (A03459 tissue IHC caption). Compare the distribution with reported high staining in small intestine endocrine cells and with a tissue cell population reported as undetected, such as adipocytes in adipose tissue (HPA tissue IHC). Diffuse staining across both contexts warrants a control review (standard IHC interpretation).
How should I score DAB1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the target cell population and staining compartment before scoring, then use the same criteria for every section (standard IHC quantification practice). For cell based staining, report the percentage of positive cells and an H score based on intensity; for discrete positive cells, report density per mm² of evaluable tissue (standard IHC quantification practice). Normalize counts to the number of relevant cells or measured viable tissue area, and keep exposure, counterstain, and DAB development consistent (standard IHC quantification practice). Record cytoplasmic and membrane associated signal separately when assessing small intestine, where both patterns are reported (HPA tissue IHC profile). Include matched controls and document excluded folds, edges, and damaged areas (standard IHC quantification practice).
How can I distinguish credible DAB1 staining from an artefact?
Judge signal by compartment and cell context: DAB1 is cytoplasmic without a transmembrane segment, while small intestine IHC reports cytoplasmic and membranous expression (UniProt O75553 subcellular annotation; UniProt O75553 topology; HPA tissue IHC profile). High staining in small intestine endocrine cells or duodenal enterocyte microvilli has tissue evidence; equally strong signal in a population reported as undetected needs closer control review (HPA tissue IHC positive and negative profiles). Reject staining confined to section edges, folds, or necrotic areas when evaluating specific signal (standard IHC interpretation). Check the no primary control and peroxidase block for persistent DAB deposits attributable to detection chemistry (standard chromogenic IHC practice). Interpret isolated nuclear staining cautiously against the cytoplasmic annotation (UniProt O75553 subcellular annotation).
Boster reagents
Best DAB1 / Disabled homolog 1 IHC Antibodies
A03459 has an IHC image from paraffin-embedded human small intestine (catalog image caption) and lists IF, IHC, and Human, Mouse, and Rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-Dab1 (Q226) Antibody
Cat # A03459
A03459 is the sole card: its IHC image shows paraffin-embedded human small intestine at 1:100 (catalog image caption). It lists IHC and IF for Human, Mouse, and Rat (catalog applications/reactivity), but has no IF image in the payload (catalog IF image alts).
Which to pick: Choose A03459 for paraffin-section IHC; its own image caption documents human small intestine at 1:100, but does not report the fixative (catalog image caption). For IF, A03459 lists total DAB1, while A03459Y232 lists phospho-DAB1 (Y232); neither has an IF image in the payload, and ICC is not listed for either (catalog titles/applications/IF image alts). Both list Human, Mouse, and Rat reactivity, but only A03459 lists IHC (catalog applications/reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75553 (DAB1_HUMAN, Disabled homolog 1).
- Human Protein Atlas. DAB1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DAB1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. DAB1 antibody validation summary (3 antibodies).
- Ephrin‑b3 modulates hippocampal neurogenesis and the reelin signaling pathway in a pilocarpine‑induced model of epilepsy. International journal of molecular medicine 2018 — PMC5881691.
- Reelin expression in human liver of patients with chronic hepatitis C infection. European journal of histochemistry : EJH 2017 — PMC5365015.
- Combinations of Low-Frequency Genetic Variants Might Predispose to Familial Pancreatic Cancer. Journal of personalized medicine 2021 — PMC8305658.
- Evidence for ApoE receptor 2-Disabled homolog-1 pathway disruption in the amygdala in sporadic Alzheimer's disease. medRxiv : the preprint server for health sciences 2025 — PMC12191094.
- PubMed PMID:9790777 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DAB1 (Disabled homolog 1). Expected localisation Cytoplasmic and membranous in small intestine (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A03459 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human small intestine tissue; fixative not specified (datasheet A03459); 2. Fixation: Image fixative and duration unreported (datasheet A03459); 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-DAB1, 1:50-1:200 (datasheet A03459); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Brain expression may not predict intestinal IHC staining (UniProt; HPA tissue IHC); Isoform / epitope: Six isoforms; epitope coverage is unknown (UniProt); Regulation: Expression regulation unspecified (UniProt). Evidence: UniProt O75553, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A03459, PMC5365015, PMC12191094. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DAB1/immunohistochemistry-ihc-guide.