DAB2 / Disabled homolog 2 · IHC design guide

Design Immunohistochemistry for DAB2

Plan DAB2 paraffin-section IHC around its reported cytoplasmic tissue staining (HPA tissue IHC). Use high-staining Hofbauer cells and lung macrophages to assess the expected pattern, and score mixed tissues by cell type (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for DAB2 (IHC for DAB2): expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A01546-3, validated IHC image, and IHC protocol steps — Printable **DAB2** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A01546-3, controls and protocol steps. Open the full DAB2 IHC guide →

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DAB2 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 tissue staining (HPA tissue IHC)
	Staining pattern	Cytoplasmic signal, mainly in placenta and peripheral leukocytes (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A01546-3)
	Positive control ⓘ	Lung+4 more · see all
	Negative control ⓘ	Appendix+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific)
	Caveat	High macrophage staining can complicate cell-level scoring (HPA tissue IHC)
	Regulation	Expression regulation is unreported (UniProt)
	Isoform / epitope	3 isoforms; cytoplasmic protein with no transmembrane segment; epitope coverage unknown (UniProt)

Section 1

Recommended DAB2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet A01546-3) is accompanied by published DAB2 IHC methods for endometrial cancer (PMC5047385) and breast cancer (PMC3956763).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human melanoma tissue; fixative not specified (datasheet A01546-3)
Fixation	Image fixative and duration unreported (datasheet A01546-3); 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 A01546-3); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A01546-3)
Primary antibody	Rabbit anti-DAB2, 2-5 μg/ml (datasheet A01546-3)
Primary incubation	Overnight at 4 °C (datasheet A01546-3)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A01546-3)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	DAB2-positive staining in macrophages of lung (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in placenta and peripheral leukocytes. No signal in the no-primary control.

💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A01546-3). Citrate microwave retrieval was reported with a different DAB2 antibody (PMC5047385 methods).

Section 2

What Is the Expected DAB2 Staining Pattern?

DAB2 is cytoplasmic and associates with clathrin-coated pits and vesicles; it has no transmembrane segment (UniProt P98082: localization and topology). In tissue IHC, expect cytoplasmic staining, especially in lung macrophages and placental Hofbauer cells (HPA: High in both). HPA rates the tissue IHC profile Enhanced, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).

What am I looking at on my slide?
Cytoplasmic staining in lung macrophages or placental Hofbauer cells, stronger than surrounding unstained cells (HPA: High in both cell types).	This matches the reported cell-specific IHC pattern (HPA: tissue IHC). Vesicle-associated or peripheral emphasis is plausible given DAB2's association with coated pits and vesicles, but chromogenic IHC may not resolve individual vesicles (UniProt P98082: localization; standard IHC resolution).
Predominantly nuclear staining in a putative positive cell, without a convincing cytoplasmic component.	Treat this as a localization mismatch requiring review, since the tissue IHC profile is cytoplasmic (HPA: tissue IHC). ICC-IF additionally reports a nucleolar fibrillar-center location; that observation alone does not establish a predominantly nuclear tissue IHC pattern (HPA: subcellular ICC-IF).
Strong signal in a cell type reported as undetected, such as bronchial basal cells or heart cardiomyocytes (HPA: Not detected in those cells).	Review cell identification and assay controls before calling it DAB2. Cross-reactivity or endogenous chromogenic activity are possible general IHC explanations; an HPA 'Not detected' entry is an observed pattern, not proof that every specimen must be negative (HPA: tissue IHC; standard IHC practice).
Uniform brown haze across cells, stroma, and empty regions, obscuring cell boundaries.	This does not support a cell-specific DAB2 call. Assess background from detection reagents, incomplete blocking, or excessive primary-antibody concentration using appropriate controls (standard IHC practice); HPA instead describes cytoplasmic expression mainly in placenta and peripheral leukocytes (HPA: tissue IHC profile).
No staining in lung macrophages or placental Hofbauer cells (HPA: High in both cell types).	An absent signal in an expected-positive cell warrants checking the assay and tissue identity before interpreting other sections as negative (standard IHC practice). The HPA profile has medium staining-to-RNA consistency, so its positive examples are useful benchmarks rather than guaranteed results in every specimen (HPA: tissue IHC reliability).

💡Expected DAB2 appearanceCall a convincing positive when brown cytoplasmic signal identifies lung macrophages or placental Hofbauer cells at the reported High staining level; isolated nuclear signal or diffuse, cell-independent brown deposit is insufficient (HPA: tissue IHC; standard IHC interpretation).

How each factor affects the staining
Cell-specific reference pattern	HPA reports High staining in lung macrophages and placental Hofbauer cells, Medium in several other listed cell types, and Not detected in selected cells (HPA: tissue IHC). Score the identified cell population within its tissue rather than assigning one intensity to an entire section (standard IHC practice).
Subcellular localization	DAB2 is cytoplasmic and associates with coated vesicles and pits; it has no transmembrane segment (UniProt P98082: localization and topology). A peripheral accent can fit that biology, but a crisp continuous surface outline is not required for a positive chromogenic section (standard IHC interpretation).
Antibody validation and isoforms	HPA lists HPA028888 as IHC Enhanced and CAB009314 as IHC Supported; these are antibody-specific validation ratings (HPA: antibodies). UniProt lists three isoforms, but the supplied record does not map either antibody's epitope to them, so isoform-specific staining cannot be inferred (UniProt P98082: isoforms).
IF/ICC Q&A: where should fluorescence localize?	Mainly to vesicles and plasma membrane, with an additional nucleolar fibrillar-center location (HPA: subcellular ICC-IF). That cell-imaging result helps assess localization; it does not replace the cytoplasmic tissue IHC reference pattern (HPA: tissue IHC and subcellular ICC-IF).
Retrieval and chromogenic detection	These are general IHC workflow variables that can alter visibility or background (standard IHC practice). The supplied sources give no DAB2-specific retrieval condition or fixation sensitivity; do not attribute an unexpected result to either mechanism without assay-specific evidence.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected-positive macrophages or Hofbauer cells are blank (HPA: High in those cells).	Possible tissue-identification or assay failure; the supplied sources do not identify a DAB2-specific failure mechanism.	Confirm the cell population and run an expected-positive section alongside a reagent control; review retrieval, antibody incubation, and detection steps as general IHC checks (standard IHC practice).
Signal appears mainly nuclear in tissue IHC.	The distribution conflicts with HPA's cytoplasmic tissue profile, although HPA notes an additional nucleolar location in ICC-IF (HPA: tissue IHC and subcellular ICC-IF).	Check morphology, counterstain, and control sections before scoring. Record a nuclear-only pattern as unresolved rather than equating it with the ICC-IF observation (standard IHC interpretation).
Broad brown staining appears in nominally negative cells or across the section.	Cross-reactivity, endogenous peroxidase activity, or reagent background are possible general chromogenic IHC causes (standard IHC practice).	Compare a no-primary control, check peroxidase blocking and washing, and reassess primary-antibody concentration; verify the scored cell type against HPA's cell-level tissue entries (standard IHC practice; HPA: tissue IHC).
A reported Not detected cell type shows a discrete positive signal (HPA: tissue IHC).	Cell misclassification or assay-specific signal is possible; an HPA absence call does not establish universal biological absence (HPA: tissue IHC; standard IHC interpretation).	Confirm histology and compare controls, then describe the result as differing from the HPA reference pattern if it persists (HPA: tissue IHC; standard IHC reporting).
Cell-specific staining is present but too faint to score confidently.	Staining intensity can vary with general IHC workflow conditions; HPA reports medium consistency between staining and RNA data (standard IHC practice; HPA: reliability).	Compare an expected-positive section processed in the same run, then review retrieval, incubation, detection, and counterstain without assuming a DAB2-specific fixation effect (standard IHC practice).
Two antibodies yield different compartments or cell populations.	Their HPA IHC validation ratings differ: HPA028888 is Enhanced and CAB009314 is Supported; the supplied evidence does not explain any particular discrepancy (HPA: antibodies).	Compare each result with the HPA cell-specific pattern and appropriate controls; report the antibody ID and unresolved difference instead of merging discordant signals into one DAB2 score (HPA: tissue IHC; standard IHC reporting).

Source: UniProt P98082 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing DAB2 staining

IHC analysis of DAB2 using anti-DAB2 antibody (A01546-3). DAB2 was detected in a paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-DAB2 Antibody (A01546-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A01546-3 · IHCIHC analysis of DAB2 using anti-DAB2 antibody (A01546-3). DAB2 was detected in a paraffin-embedded section of human melanoma tissue. Heat me

IF analysis of DAB2 using anti-DAB2 antibody (A01546-3). DAB2 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-DAB2 Antibody (A01546-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A01546-3 · IFIF analysis of DAB2 using anti-DAB2 antibody (A01546-3). DAB2 was detected in an immunocytochemical section of Hela cells. Enzyme antigen re

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000153071-DAB2/tissue/cerebral+cortex#imid_16546459

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000153071-DAB2/tissue/colon#imid_16540169

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000153071-DAB2/tissue/liver#imid_16540162

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000153071-DAB2/tissue/kidney#imid_16540163

HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA028888; green = DAB2, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for DAB2 IHC & IF

🧪Run lung first and score macrophages, where DAB2 staining is High (HPA: High in lung macrophages). Use appendix endocrine cells as the negative tissue control (HPA: Not detected in appendix endocrine cells); on the lung slide, use neighboring unstained cells as a background reference without assuming all nonmacrophage cells are DAB2-negative.

Positive control tissue: Lung (Macrophages, HPA High)

Negative control tissue: Appendix (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show DAB2 in CACO-2, HeLa, U2OS, NIH 3T3, with annotated localisation: Vesicles (enhanced), Plasma membrane (enhanced) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) control, host-matched rabbit IgG isotype control, and DAB2-knockout biological negative (selected caption: rabbit anti-DAB2; standard IHC practice). Block endogenous peroxidase and check the lung macrophage field for pigment that could be mistaken for DAB signal (standard chromogenic IHC practice).

⚠️Feasibility: The selected A01546-3 paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but its fixative is unreported; no target-specific fixation window or fixation effect is supplied (selected tissue-IHC caption). Retrieval is part of that demonstrated IHC procedure, although the evidence does not establish whether DAB2 staining depends on it (selected tissue-IHC caption). The evidence does not establish that frozen sections or IF are easier; inspect lung macrophage pigment when scoring DAB staining (selected tissue-IHC caption; standard chromogenic IHC practice).

HPA tissue IHC evidence for DAB2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Lung	Macrophages	High	Protein (IHC)	HPA →
Placenta	Hofbauer cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Medium	Protein (IHC)	HPA →
Cerebellum	Purkinje cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Appendix	Endocrine cells	Not detected	Protein (IHC)	HPA →
Bronchus	Basal cells	Not detected	Protein (IHC)	HPA →
Duodenum	Endocrine cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Seminal vesicle	Glandular cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced DAB2 IHC Tips

Troubleshoot DAB2 staining in paraffin sections by checking retrieval, cell identity, subcellular pattern, and controls before comparing signal across samples.

Which retrieval condition should I use when DAB2 staining is weak?

Start with heat-mediated EDTA at pH 8.0 for this catalog antibody in paraffin sections (datasheet A01546-3). The selected tissue image used that retrieval before 2 μg/mL primary antibody overnight at 4°C, followed by peroxidase detection and DAB (caption A01546-3). Keep heating, cooling, and section thickness consistent across cases, then compare a lung macrophage-rich area with a no-primary control (standard IHC practice; HPA: high in lung macrophages). If staining remains weak, optimize retrieval duration in a small series before testing another buffer, because an alternative condition lacks product-specific support here (datasheet A01546-3; standard IHC practice).

Does the tissue image establish which fixative preserves DAB2 staining best?

The selected paraffin-section image does not report a fixative, so DAB2-specific fixation sensitivity is unknown for this antibody (caption A01546-3). Record the fixative and processing history for each block, and compare blocks only after checking section age, retrieval, and detection conditions (standard IHC practice). If signal varies between specimens, use similarly processed control tissue in the same run to assess whether the difference tracks processing (standard IHC practice). Do not attribute weak staining to a particular fixative from the tissue expression profile or DAB2 topology; neither establishes fixation sensitivity (HPA: tissue IHC profile; UniProt P98082 topology).

Should DAB2 stain the membrane, cytoplasm, or both in tissue sections?

Expect cytoplasmic staining with vesicular or membrane-associated enrichment: DAB2 associates with clathrin-coated vesicles and pits and has no transmembrane segment (UniProt P98082). Cell imaging also places DAB2 mainly at vesicles and the plasma membrane, although that pattern needs assessment in each tissue section (HPA: subcellular summary; standard IHC practice). Score signal within identified cells because lung macrophages and placental Hofbauer cells can stain highly, while neighboring cell populations may differ (HPA: tissue IHC). During mitosis, DAB2 can shift from membrane to cytoplasm, so a cytoplasmic pattern alone does not establish mislocalisation (UniProt P98082).

Could isoforms or epitope changes explain discordant DAB2 staining?

DAB2 has 3 listed isoforms and a PID domain spanning residues 45–196, but the supplied product caption does not map this antibody’s epitope (UniProt P98082; caption A01546-3). Therefore, tissue staining cannot identify a specific isoform or establish equal recognition of all isoforms without epitope information (UniProt P98082 isoforms; standard IHC interpretation). Document the immunogen or mapped epitope before comparing antibodies, and check whether it lies in a region shared by the isoforms (standard antibody validation practice). Reported modifications include phosphotyrosine 170 and phosphoserine 193; their effect on this antibody’s staining is untested here (UniProt P98082; caption A01546-3).

How can IF help resolve an ambiguous DAB2 IHC pattern?

Use IF/ICC as a separately validated follow-up when chromogenic staining cannot assign DAB2 signal to a cell or compartment (standard IHC/IF practice). Multiplex DAB2 with a validated macrophage marker when examining lung macrophages, and include single-label controls to assess channel bleed-through (HPA: high in lung macrophages; standard IF practice). Choose a fluorophore channel after checking tissue autofluorescence, particularly when weak vesicular signal must be distinguished from diffuse background (HPA: vesicular location; standard IF practice). Because DAB2 lacks a transmembrane segment and its antibody epitope is unmapped here, optimize permeabilisation for intracellular access without assuming the epitope’s accessibility (UniProt P98082 topology; caption A01546-3).

How do I separate nonspecific DAB signal from DAB2 staining?

The selected image used 10% goat serum before primary incubation and peroxidase-linked detection with DAB (caption A01546-3). Include a no-primary section and an appropriate peroxidase block to assess secondary reagent binding and endogenous enzyme signal (standard IHC practice). In the pictured melanoma tissue, inspect unstained or no-primary material for native pigment before assigning dark deposits to DAB2 (caption A01546-3; standard histology practice). Judge remaining signal by its distribution within intact cells and compare it with expected cytoplasmic, vesicular, or membrane-associated localisation (UniProt P98082; HPA: subcellular summary; standard IHC practice).

How should I quantify DAB2 when positive cell types vary across sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and tissue region before scoring, because DAB2 staining can be high in lung macrophages and placental Hofbauer cells (HPA: tissue IHC). For an identified population, record the percentage of positive cells and intensity, or calculate an H-score from intensity categories with a 0–300 range (standard IHC scoring practice). For dispersed positive cells, report their density per mm² of viable tissue and the number of cells assessed (standard quantitative IHC practice). Normalize comparisons to the same cell type and viable area, using consistent staining and image thresholds across cases (standard quantitative IHC practice).

What findings make a DAB2-positive IHC result convincing?

A convincing result places signal in intact, identifiable cells with a cytoplasmic, vesicular, or membrane-associated pattern and passes a no-primary control (UniProt P98082; HPA: subcellular summary; standard IHC practice). High staining in lung macrophages or placental Hofbauer cells is compatible with the tissue profile; unexpected cell types need independent confirmation (HPA: tissue IHC; standard IHC practice). Treat isolated section-edge deposits, necrotic areas, or signal persisting without primary antibody as possible artefact, including endogenous peroxidase activity (standard IHC practice). Predominantly nucleolar staining warrants review rather than automatic rejection, because a nucleolar location is additionally reported in cell imaging (HPA: subcellular summary).

Boster reagents

Best DAB2 / Disabled homolog 2 IHC Antibodies

The catalog antibody has IHC data from a human melanoma paraffin section and IF/ICC data from HeLa cells (catalog image captions); listed reactivity covers human, mouse, rat and monkey (catalog reactivity).

Real IHC data

Anti-DAB2 Antibody ®

Cat # A01546-3

View product →

A01546-3 will render with its IHC figure from a human melanoma paraffin section (A01546-3 IHC image caption). The same SKU lists IF/ICC and has a separate HeLa cell IF image (A01546-3 applications; A01546-3 IF image caption).

Which to pick: Choose A01546-3 for paraffin-section IHC: its own caption shows heat retrieval in EDTA at pH 8.0 and staining at 2 μg/ml; the fixative is unreported (A01546-3 IHC image caption). For IF/ICC, the same SKU has a HeLa cell image, and its listed IF concentration is 5 μg/ml (A01546-3 IF image caption; catalog dilution). For cross-species work, A01546-3 lists human, mouse, rat and monkey reactivity, although its supplied IHC and IF images show human samples only; clonality is unreported (catalog reactivity; A01546-3 image captions; catalog clone field).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry P98082 (DAB2_HUMAN, Disabled homolog 2).
Human Protein Atlas. DAB2 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. DAB2 subcellular location (ICC-IF): Mainly localized to the plasma membrane and vesicles. In addition localized to the nucleoli fibrillar center..
Human Protein Atlas. DAB2 antibody validation summary (2 antibodies).
Expression Pattern of DAB Adaptor Protein 2 in Left- and Right-Side Colorectal Carcinoma. Genes 2023 — PMC10379130.
Expression and clinical significance of the transforming growth factor-β signalling pathway in endometrial cancer. Histopathology 2011 — PMC5047385.
Analysis of the role of Igf2 in adrenal tumour development in transgenic mouse models. PloS one 2012 — PMC3429465.
Loss of Dab2 expression in breast cancer cells impairs their ability to deplete TGF-β and induce Tregs development via TGF-β. PloS one 2014 — PMC3956763.
PubMed PMID:8660969 — UniProt-cited evidence.
PubMed PMID:9620555 — UniProt-cited evidence.
PubMed PMID:10340382 — UniProt-cited evidence.