This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
DAB1 / Disabled homolog 1 · IHC design guide
Design Immunohistochemistry for DAB1
Plan DAB1 IHC in paraffin sections using small intestine as a strong-staining reference (HPA tissue IHC). The guide covers expected staining, fixation consistency, antibody dilution, and interpretation; the catalog antibody lists an IHC dilution 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 (UniProt); intestinal membranous (HPA tissue IHC) | |
| Staining pattern | Intestinal microvilli stain strongly; cytoplasmic and membranous pattern (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 fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | CNS staining is low despite brain-enhanced RNA (HPA tissue IHC) | |
| Regulation | Tissue-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | Six splice isoforms; check epitope coverage (UniProt) |
Section 1
Recommended DAB1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by four article-derived DAB1 IHC protocols, including one for phospho-DAB1 Tyr232 (PMC5881691 methods).
| 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 antigen-retrieval rule); compare staining with appropriate controls.
Section 2
What Is the Expected DAB1 Staining Pattern?
DAB1 is a cytoplasmic protein with no transmembrane segment (UniProt O75553 topology). In paraffin-section IHC, expect cytoplasmic and membranous staining in small intestine, especially duodenal enterocyte microvilli and intestinal endocrine cells; selected neuronal populations also stain (HPA: tissue IHC). HPA rates the tissue pattern Enhanced, citing high consistency between antibody staining and RNA expression (HPA: reliability).
| Strong staining along duodenal enterocyte microvilli, with staining of small-intestinal endocrine cells. | This matches HPA's high staining in both cell populations and its cytoplasmic and membranous small-intestine profile (HPA: tissue IHC). Assess the cellular pattern alongside the section's morphology; intensity alone does not establish specificity (general IHC practice). |
| Staining in cerebellar molecular-layer cells, cortical neurons or retinal ganglion cells is weaker than in the intestinal positive control. | These populations are reported at medium staining, while HPA describes low expression across CNS and retina overall (HPA: tissue IHC). Compare the same named cell populations, rather than treating an entire brain or retinal section as uniformly positive (HPA: tissue IHC). |
| A strong, predominantly nuclear signal replaces the expected cytoplasmic pattern. | UniProt places DAB1 in the cytoplasm, and HPA describes intestinal staining as cytoplasmic and membranous (UniProt O75553: subcellular location; HPA: tissue IHC). Treat isolated nuclear staining as suspect; inspect the counterstain and detection controls before scoring it as DAB1 (general IHC practice). |
| Strong staining appears in a cell population HPA lists as undetected, such as adipocytes in adipose tissue. | HPA reports no detectable staining in those adipocytes (HPA: tissue IHC). Check tissue identity and cell morphology, then investigate antibody cross-reactivity or endogenous detection activity with appropriate controls (general IHC practice). An unexpected result warrants validation rather than automatic rejection. |
| No signal appears in duodenal enterocyte microvilli or small-intestinal endocrine cells. | Both are high-staining populations in HPA's tissue record (HPA: tissue IHC). Verify that the expected cells are present, then check primary-antibody and detection controls and the retrieval conditions used for the IHC assay (general IHC practice). A blank section alone cannot establish absent DAB1. |
💡Expected DAB1 appearanceCall a section positive when the expected cells show a convincing cytoplasmic or microvillar membranous pattern—high in duodenal enterocyte microvilli or small-intestinal endocrine cells, medium in the specified neural populations—while isolated strong nuclear staining or signal in HPA-undetected cells prompts a specificity check (HPA: tissue IHC; UniProt O75553: subcellular location).
| Cellular location and topology | DAB1 is cytoplasmic and has no transmembrane segment (UniProt O75553: subcellular location and topology). HPA nevertheless reports membranous intestinal staining, including high signal at enterocyte microvilli (HPA: tissue IHC). Interpret that observed pattern in its cell context; it does not imply that DAB1 spans a membrane. |
| Choice of reference tissue and cells | Duodenal enterocyte microvilli and small-intestinal endocrine cells are high-staining references; cerebellar molecular-layer cells, cortical neurons and retinal ganglion cells are medium (HPA: tissue IHC). HPA lists several undetected cell populations, including adipocytes in adipose tissue and endocrine cells in colon (HPA: tissue IHC). Match the named cells when comparing sections. |
| Evidence tied to the antibody | HPA marks IHC staining Enhanced for HPA052033 and CAB032329; its Enhanced tissue profile reflects consistency with RNA expression (HPA: antibodies; HPA: reliability). HPA067495 has Enhanced ICC validation but no IHC status in the supplied antibody record (HPA: antibodies). Do not transfer an ICC validation label to paraffin-section IHC. |
| Isoforms and antibody epitope | UniProt lists six DAB1 isoforms and a PID domain at residues 36–189 (UniProt O75553: isoforms and domains). The supplied record gives no epitope for the catalog antibody, so these facts cannot predict which isoforms it detects or whether retrieval exposes its epitope. Review antibody-specific IHC documentation when interpreting an unexpected pattern (general IHC practice). |
| IF/ICC interpretation | What should IF/ICC show? HPA reports an enhanced vesicular location and images from HeLa and RT-4 cells (HPA: subcellular ICC-IF). That observation can guide interpretation of an IF image, but it does not replace the tissue-specific IHC pattern or establish an IF protocol (HPA: tissue IHC; HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| The intestinal positive control is blank. | The slide may lack the named positive cells, or an IHC reagent or retrieval step may have failed (general IHC practice). HPA expects high staining in duodenal enterocyte microvilli and small-intestinal endocrine cells (HPA: tissue IHC). | Confirm cell identity and section integrity, then check the assay's primary-antibody, retrieval and detection controls (general IHC practice). No DAB1-specific fixation sensitivity is reported in the supplied sources. |
| The slide shows strong nuclear staining. | The compartment conflicts with UniProt's cytoplasmic location and HPA's intestinal cytoplasmic and membranous profile (UniProt O75553: subcellular location; HPA: tissue IHC). Counterstain overlap or nonspecific detection may confuse interpretation (general IHC practice). | Inspect the counterstain separately, compare a known positive section and check a primary-antibody omission control (general IHC practice). Score DAB1 only where the cell and compartment pattern is supported. |
| An HPA-undetected cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA lists, for example, adipose-tissue adipocytes as undetected (HPA: tissue IHC). | Verify the cell type, examine a primary-antibody omission control and check the detection-system controls (general IHC practice). Record any reproducible departure from HPA's pattern rather than assigning it to DAB1 solely by color. |
| Diffuse brown haze obscures cell boundaries. | Nonspecific background or endogenous chromogenic detection activity can reduce contrast in IHC (general IHC practice). HPA's named-cell staining pattern cannot be assessed reliably through haze (HPA: tissue IHC). | Review blocking, wash and detection controls and adjust assay conditions according to the antibody's IHC instructions (general IHC practice). Reassess microvilli and endocrine cells only after cellular boundaries are interpretable (HPA: tissue IHC). |
| Neural staining seems faint beside a strong intestinal control. | HPA rates the specified cerebellar, cortical and retinal cells medium, while the two intestinal reference populations are high (HPA: tissue IHC). HPA also summarizes overall CNS and retinal expression as low (HPA: tissue IHC). | Compare the named neural cells with local background and their morphology; avoid requiring intestinal-level intensity in every neural cell (HPA: tissue IHC). Use the high-staining intestinal cells to check that the assay worked. |
| An antibody with ICC evidence gives an unexpected IHC pattern. | ICC and IHC validation are application specific (general IHC practice). HPA067495 has Enhanced ICC status but no supplied IHC status, whereas HPA052033 and CAB032329 have Enhanced IHC status (HPA: antibodies). | Check the antibody identifier and its IHC validation before interpreting the paraffin section (HPA: antibodies). Compare the result with HPA's tissue and cell pattern, using appropriate IHC controls (HPA: tissue IHC; general IHC 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
🧪Start with small intestine: endocrine cells should stain (HPA: High in small-intestinal endocrine cells; selected A03459 caption: paraffin-embedded human small intestine at 1:100), and run adipose tissue as a negative, focusing on adipocytes (HPA: Not detected in adipocytes). On the positive slide, use nearby non-endocrine cells as candidate internal negatives: expect background-level staining, but verify it with the no-primary control because their DAB1 status is not specified by the HPA row (HPA: High is specified for endocrine cells).
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, an isotype control matched to the primary antibody’s host species, immunoglobulin class and clonality, and DAB1-knockout material as a specificity control (standard IHC practice). Quench endogenous peroxidase in small-intestine sections and check endogenous biotin background if detection uses biotin (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected A03459 paraffin-section caption does not state a fixative (selected A03459 tissue-IHC caption). Antigen-retrieval dependence and whether frozen sections or IF are easier are unreported; optimize retrieval empirically for paraffin IHC (standard IHC practice), while ICC-IF images show enhanced vesicular localization in HeLa and RT-4 (HPA subcellular). Endogenous peroxidase or biotin in small-intestine sections can complicate chromogenic interpretation (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 using the documented tissue pattern, the selected antibody image, and standard chromogenic IHC controls.
How should I retrieve DAB1 in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat induced retrieval at 95–98 °C for 20 min (page retrieval setting). Cool sections consistently, then compare the same antibody dilution and detection conditions across runs so retrieval is the main variable (standard IHC practice). The selected antibody image shows staining in paraffin embedded human small intestine at 1:100, but its caption does not specify retrieval conditions (A03459 caption). Include small intestine with identifiable enterocytes and endocrine cells as a reference, and review staining by cell type rather than judging overall section darkness (HPA tissue IHC; standard IHC practice).
Could fixation explain inconsistent DAB1 staining between paraffin blocks?
Target specific sensitivity to fixation is unknown because the selected antibody caption identifies paraffin embedded tissue but does not state the fixative or fixation duration (A03459 caption). Record the fixative, duration, tissue thickness, and processing history for each block before comparing staining intensity (standard IHC practice). If material permits, stain blocks with different processing histories in the same run, using identical retrieval and detection steps and a small intestine reference section (HPA tissue IHC; standard IHC practice). Treat differences as processing associated observations until matched controls establish their cause; neither the tissue staining pattern nor DAB1 phosphorylation identifies a fixation effect (HPA tissue IHC; UniProt O75553).
Where should DAB1 staining appear in a convincing positive section?
Assess cellular staining in context: DAB1 is annotated as cytoplasmic and has no transmembrane segment (UniProt O75553). Tissue IHC reports cytoplasmic and membranous expression in small intestines, with high staining in duodenal enterocyte microvilli and small intestine endocrine cells (HPA tissue IHC). A separate subcellular IF profile assigns DAB1 to vesicles, so a fine intracellular pattern can be plausible without establishing the identity of every stained structure (HPA subcellular). Compare candidate signal with the expected cell boundaries and a counterstain; isolated nuclear staining or diffuse extracellular deposits warrant review of controls and morphology before being called DAB1 (standard IHC practice; UniProt O75553).
Could an epitope or DAB1 isoform explain different staining patterns?
DAB1 has 6 listed isoforms, and the supplied evidence does not map this antibody's epitope to any of them (UniProt O75553; A03459 caption). Its PID domain spans residues 36–189, while listed modified residues include phosphotyrosines at 198, 220, and 232 and CDK5 associated phosphoserine at 524 (UniProt O75553). These annotations justify checking the antibody's documented immunogen and epitope before interpreting a difference as isoform or phosphorylation specific (UniProt O75553; standard IHC practice). Keep retrieval, dilution, and detection matched when comparing sections, and report uncertain epitope coverage instead of assigning a stained population to one splice form (standard IHC practice).
How can IF help assess DAB1 localisation alongside cell identity?
For IF or ICC, pair DAB1 with a marker for the cell population being examined, such as a validated neuronal marker when assessing the reported cortical neuronal signal (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence, favoring a well separated longer wavelength channel when background is high, and acquire single stain controls for multiplex interpretation (standard IF practice). Because DAB1 is cytoplasmic without a transmembrane segment, use a controlled permeabilisation step when the antibody must reach an intracellular epitope; the precise epitope is not supplied (UniProt O75553; A03459 caption). Compare puncta with the reported vesicular IF localisation while retaining cell boundaries and channel controls in the review (HPA subcellular; standard IF practice).
What should I check when DAB1 chromogenic staining looks diffuse?
Inspect a no primary control and the tissue morphology before increasing the stringency of washes or blocking (standard IHC practice). In a peroxidase and DAB workflow, include an endogenous peroxidase block and check whether residual pigment appears in the no primary control (standard IHC practice). Titrate the primary antibody around the image's reported 1:100 use in paraffin embedded small intestine, changing one condition at a time; that caption documents an image, not an optimized range for every specimen (A03459 caption; standard IHC practice). Judge improvement by preservation of identifiable enterocyte or endocrine cell staining rather than by a lower whole section background alone (HPA tissue IHC; standard IHC practice).
How should I quantify DAB1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, because reported DAB1 staining varies among small intestine cell types and brain regions (HPA tissue IHC). For cellular signal, record the percentage of positive cells and an H-score from intensity categories; for a defined sparse population, positive cells per mm² can be useful (standard IHC practice). Normalize counts to the number of eligible cells or the measured tissue area, and keep section thickness, imaging, and detection settings consistent across samples (standard IHC practice). Exclude folds, torn edges, and necrotic areas by a prespecified rule, then report the compartments scored and the threshold established with control sections (standard IHC practice).
How do I distinguish a true DAB1 signal from staining artefact?
Look for reproducible cellular staining in the reported small intestine populations, including enterocyte microvilli and endocrine cells, while recognizing that some neuronal populations show lower reported signal (HPA tissue IHC). DAB1 is cytoplasmic with no transmembrane segment, and an IF profile reports vesicles; isolated nuclear deposits should therefore prompt closer review rather than an immediate positive call (UniProt O75553; HPA subcellular). Compare staining with a no primary control and examine edges, folds, and necrotic areas for nonspecific deposition (standard IHC practice). In peroxidase and DAB detection, residual endogenous enzyme activity can imitate positive chromogen, so verify the peroxidase block and require signal within intact, identifiable cells (standard IHC practice).
Boster reagents
Best DAB1 / Disabled homolog 1 IHC Antibodies
A03459 has a human paraffin-section IHC figure (catalog image caption) and listed IF use; its reported reactivity covers human, mouse, and rat (catalog: applications and reactivity).
Real IHC data
Anti-Dab1 (Q226) Antibody
Cat # A03459
Only A03459 renders an IHC card: its figure shows paraffin-embedded human small intestine at 1:100 (catalog image caption). IF is listed for A03459, but no IF figure is supplied (catalog: applications and image alts).
Which to pick: Choose A03459 for tissue IHC: its own figure supports paraffin-embedded human small intestine, with the fixative unreported (catalog image caption). For IF, A03459 and A03459Y232 both list human, mouse, and rat reactivity; A03459Y232 targets phospho-DAB1 Y232, while neither has an IF figure or an explicit ICC listing (catalog: titles, applications, reactivity, and image alts).
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 (UniProt); intestinal membranous (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 fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: CNS staining is low despite brain-enhanced RNA (HPA tissue IHC); Isoform / epitope: Six splice isoforms; check epitope coverage (UniProt); Regulation: Tissue-dependent expression (HPA tissue IHC). Evidence: UniProt O75553, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A03459, PMC5881691, PMC5365015. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DAB1/immunohistochemistry-ihc-guide.