DAB2 / Disabled homolog 2 · IHC design guide

Design Immunohistochemistry for DAB2

Plan DAB2 staining in paraffin sections using the catalog antibody’s IHC conditions (datasheet A01546-3). Compare cytoplasmic staining with the tissue profile, paying particular attention to macrophages (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 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 sections. (standard IHC practice; not target-specific)
	Caveat	Macrophage signal may complicate cell attribution (HPA tissue IHC)
	Regulation	Staining intensity regulators unreported (UniProt)
	Isoform / epitope	3 isoforms; confirm epitope coverage (UniProt)

Section 1

Recommended DAB2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is supplemented by published DAB2 protocols for endometrial cancer and mouse adrenal sections (PMC5047385; PMC3429465).

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 heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01546-3); the published protocols used citrate retrieval with their respective antibodies (PMC5047385; PMC3429465).

Section 2

What Is the Expected DAB2 Staining Pattern?

DAB2 is a cytoplasmic adaptor associated with clathrin-coated vesicles and pits; 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 its tissue staining evidence Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Strong cytoplasmic staining in lung macrophages or placental Hofbauer cells.	This matches the clearest supplied tissue examples (HPA: High in both cell types). Score the named cells; other cells in the section can have different expression (HPA tissue IHC).
Staining is confined to nuclei, with no convincing cytoplasmic signal.	That is discordant with the tissue profile and warrants an artefact check (HPA tissue IHC: cytoplasmic). Do not dismiss every nuclear signal: a nucleolar location is additionally reported in ICC-IF (HPA subcellular).
Strong staining appears in a cell type listed as not detected, such as cardiomyocytes.	Check for cross-reactivity or endogenous detection activity before calling it DAB2 (HPA: cardiomyocytes not detected; standard IHC practice). A cell-level finding does not make the whole tissue negative (HPA tissue IHC).
Diffuse color covers cells and surrounding tissue without clear cell boundaries.	This is difficult to score as DAB2 because the expected tissue pattern is cellular and cytoplasmic (HPA tissue IHC). Assess background with a no-primary control and review blocking and detection steps (standard IHC practice).
No signal appears in lung macrophages or placental Hofbauer cells.	A negative result in these known-positive cell types calls for a technical check before biological interpretation (HPA: High in both). Confirm the cells are present and review the IHC controls (standard IHC practice).

💡Expected DAB2 appearanceCall a convincing positive when macrophages or Hofbauer cells show clear cytoplasmic staining, potentially strong (HPA: High); isolated nuclear staining or uniform tissue-wide color needs investigation (HPA tissue IHC; standard IHC practice).

How each factor affects the staining
Compartment and slide resolution	DAB2 associates with cytoplasm and coated vesicles or pits (UniProt P98082). HPA reports vesicles and plasma membrane by ICC-IF; fine puncta may be hard to resolve in chromogenic sections (HPA subcellular; standard IHC practice).
Cell-specific reference levels	Lung macrophages and Hofbauer cells are High; several other listed cell types are Medium, Low or not detected (HPA tissue IHC). Interpret the identified cell type rather than assigning one level to an entire tissue.
Antibody validation	HPA028888 has Enhanced IHC validation; CAB009314 has Supported IHC validation (HPA antibodies). These ratings inform confidence in the reported pattern but do not validate an unspecified catalog antibody (HPA antibodies).
Isoform coverage	DAB2 has 3 listed isoforms (UniProt P98082). The supplied record does not identify the catalog antibody's epitope or isoform coverage, so an isoform-specific staining difference cannot be predicted.
Q: Can ICC-IF localization be used to score IHC?	A: It can inform compartment expectations: HPA reports vesicles and plasma membrane, plus a nucleolar location, in ICC-IF (HPA subcellular). Score paraffin-section staining against the tissue IHC cell pattern (HPA tissue IHC).
Fixation sensitivity	Target-specific fixation sensitivity is unreported in the supplied UniProt and HPA evidence. If staining fails, review processing and retrieval as general IHC workflow checks (standard IHC practice), without assigning a DAB2-specific fixation effect.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive cells are present but unstained.	The result conflicts with reported High staining in lung macrophages or Hofbauer cells (HPA tissue IHC); the cause is undetermined.	Check a positive control, antibody and detection steps, and the recorded retrieval conditions (standard IHC practice). Do not infer DAB2-specific fixation sensitivity from this result.
Only nuclear staining is visible.	Nuclear-only staining differs from the predominant cytoplasmic tissue pattern (HPA tissue IHC), although ICC-IF reports an additional nucleolar location (HPA subcellular).	Review morphology and controls, then seek cytoplasmic staining in known-positive cells before accepting the IHC pattern (HPA tissue IHC; standard IHC practice).
A listed negative cell type stains strongly.	The signal may reflect cross-reactivity or endogenous detection activity (standard IHC practice); HPA reports no detection in bronchial basal cells and cardiomyocytes (HPA tissue IHC).	Verify cell identity and run a no-primary control; interpret the finding at cell level rather than treating the whole section as a negative control (standard IHC practice; HPA tissue IHC).
Broad, diffuse color obscures cell boundaries.	Background or excessive detection can prevent assessment of the cellular cytoplasmic pattern (standard IHC practice; HPA tissue IHC).	Compare with a no-primary control and review blocking, washes and detection conditions (standard IHC practice). Score DAB2 only where cell-associated signal is distinguishable (HPA tissue IHC).
Staining is weak in a Medium-level reference cell.	Medium is the reported level for several cell types, including adrenal glandular cells and endometrial stromal cells (HPA tissue IHC); weak signal alone does not identify a technical failure.	Compare like cell types and check the positive control before changing conditions (HPA tissue IHC; standard IHC practice). Use lung macrophages or Hofbauer cells as stronger references (HPA: High).
The catalog antibody gives a pattern unlike HPA images.	The supplied HPA validation applies to HPA028888 and CAB009314; it does not establish the catalog antibody's performance or epitope (HPA antibodies).	Check the catalog antibody's IHC-P validation and compare matched cell types and controls (standard IHC practice). Treat the HPA pattern as a reference, with its reported medium antibody/RNA consistency (HPA tissue IHC).

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 require staining in its macrophages (High; HPA: lung macrophages), then use seminal vesicle glandular cells as the negative comparator (HPA: Not detected in seminal vesicle glandular cells). On the lung slide, compare macrophage staining with background in morphologically distinct cells that show no signal; the HPA macrophage row does not establish that every neighboring cell is 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) slide, an isotype control matched to the primary antibody’s host species and isotype, and a DAB2 knockout specimen where available. For lung, quench endogenous peroxidase and inspect macrophage pigment before interpreting DAB signal (standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01546-3 paraffin-section caption does not state its fixative (selected IHC caption). That caption uses heat retrieval in EDTA, pH 8.0, but does not establish that retrieval is required; no matched frozen-section or tissue-IF protocol is supplied, so neither can be judged easier (selected IHC caption). Lung macrophage pigment may complicate chromogenic scoring, so assess signal against the control slides (standard 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, cellular localisation, controls and cell specific scoring.

What retrieval should I try first when DAB2 staining is weak?

Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01546-3). The selected image used this retrieval before 2 μg/ml primary antibody overnight at 4°C, so keep those conditions together for the first comparison (datasheet A01546-3). If signal remains weak, vary heating duration on adjacent sections while keeping antibody concentration, detection and imaging consistent (standard IHC practice). Check whether cytoplasmic and vesicular staining improves in the expected cells, since DAB2 occupies those compartments (UniProt P98082 subcellular location; HPA tissue IHC). Record tissue preservation and section loss alongside signal, because harsh retrieval can make an apparent gain difficult to interpret (standard IHC practice).

Could fixation explain variable DAB2 staining between paraffin blocks?

The selected paraffin section caption does not report a fixative, so target specific DAB2 sensitivity to fixation is unknown (datasheet A01546-3). Record each block’s fixative, fixation duration and processing history before comparing staining across blocks (standard IHC practice). Run matched sections with the same EDTA pH 8.0 retrieval, 2 μg/ml primary antibody and overnight incubation at 4°C used in the selected image (datasheet A01546-3). Include a known staining control in each run and judge both cellular signal and tissue morphology (standard IHC practice). A difference between blocks warrants a controlled processing comparison; the available caption cannot establish which fixation condition caused it (datasheet A01546-3).

Should DAB2 appear at the membrane, in vesicles or throughout the cytoplasm?

Assess cytoplasmic staining together with membrane associated and vesicular patterns: DAB2 is reported in cytoplasm, clathrin coated vesicles and coated pits (UniProt P98082 subcellular location). HPA places its main subcellular signal at vesicles and the plasma membrane, with an additional nucleolar fibrillar center location (HPA subcellular). Compare patterns within the same cell type and section before treating a diffuse deposit as specific staining (standard IHC practice). Mitotic cells may differ because DAB2 progressively moves from membrane to cytoplasm during mitosis (UniProt P98082 subcellular location). Use a 2 μg/ml reference section from the selected paraffin workflow to check whether altered retrieval changes the pattern as well as its intensity (datasheet A01546-3).

How can I assess whether the antibody detects the DAB2 isoform or epitope I need?

DAB2 has 3 listed isoforms, so confirm the catalog antibody’s immunogen or mapped epitope against the isoform required for the experiment (UniProt P98082 isoforms; standard IHC practice). The supplied caption establishes staining of a paraffin section at 2 μg/ml, but gives no epitope map or isoform selectivity (datasheet A01546-3). DAB2 contains a PID domain at residues 45–196 and reported modified residues including phosphotyrosine 170; these are sequence features, not evidence of antibody sensitivity to modification (UniProt P98082 domains and modified residues). Compare independently validated reagents or an appropriate loss of target control when isoform attribution matters (standard IHC practice). Score only the supported cellular pattern while that specificity remains unresolved (UniProt P98082 subcellular location; standard IHC practice).

How should I use IF to investigate an ambiguous DAB2 IHC pattern?

Use IF as a companion localisation check for the chromogenic IHC result, with a marker identifying the cell population under study (standard IF practice). A macrophage marker is useful where the IHC question concerns lung macrophages or placental Hofbauer cells, both reported as high for DAB2 (HPA tissue IHC). Choose fluorophores in channels with low tissue autofluorescence, and inspect single stain and no primary controls before interpreting overlap (standard IF practice). DAB2 has no transmembrane segment and occurs in cytoplasm and coated vesicles, so use controlled permeabilisation if the antibody epitope faces the cytosol (UniProt P98082 topology and subcellular location; standard IF practice). The paraffin IHC caption reports 2 μg/ml antibody overnight at 4°C; establish IF conditions separately rather than treating that caption as IF validation (datasheet A01546-3).

What should I check when DAB staining obscures DAB2 positive cells?

First inspect a no primary control and confirm that endogenous peroxidase was blocked before HRP and DAB development (standard IHC practice). The selected workflow used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase conjugated secondary for 30 minutes at 37°C (datasheet A01546-3). If background persists, compare primary antibody titrations and shorter chromogen development on adjacent sections while holding retrieval constant (standard IHC practice). Examine tissue edges, folds and damaged regions separately from intact cells because deposits there can mimic cellular staining (standard IHC practice). Retain a positive control when reducing signal so that a cleaner section still shows the expected DAB2 compartment (UniProt P98082 subcellular location; standard IHC practice).

How should I quantify DAB2 across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, because DAB2 expression and localisation vary by cell type and subcellular site (HPA tissue IHC; UniProt P98082 subcellular location). For a defined population, record the percentage of positive cells and an intensity weighted H-score using 0–3 intensity categories (standard IHC practice). For infiltrating cells, positive cell density per mm² can accompany the fraction of positive cells among all cells of that type (standard IHC practice). Normalise to the number of eligible cells or assessed tissue area, and exclude folds and necrotic regions under preset rules (standard IHC practice). Keep retrieval, 2 μg/ml reference antibody condition and imaging settings consistent across the comparison (datasheet A01546-3; standard IHC practice).

How do I distinguish true DAB2 staining from an IHC artefact?

Check whether staining follows DAB2’s cytoplasmic, vesicular or coated pit associated distribution in intact cells (UniProt P98082 subcellular location). Strong signal in lung macrophages or placental Hofbauer cells fits reported high expression, while staining in a different cell population needs its own control rather than automatic attribution to DAB2 (HPA tissue IHC; standard IHC practice). Treat edge staining, necrotic deposits and signal retained in a no primary control as possible artefacts, and evaluate endogenous peroxidase blocking when DAB is used (standard IHC practice). An exclusively nuclear pattern warrants scrutiny: HPA reports an additional nucleolar fibrillar center location, but its main IF locations are vesicles and plasma membrane (HPA subcellular). Compare morphology, compartment and control sections before assigning a biological difference, particularly when an antibody has only medium consistency between staining and RNA data (HPA tissue IHC).

Boster reagents

Best DAB2 / Disabled homolog 2 IHC Antibodies

A01546-3 has IHC data from a human melanoma paraffin section and IF data from HeLa cells (catalog image captions); listed reactivity is 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 caption). The same SKU is listed for IHC and IF/ICC, with IF imaging in HeLa cells (catalog applications; A01546-3 IF caption).

Which to pick: Choose A01546-3 for paraffin-section IHC: its own caption shows a human melanoma section with EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (A01546-3 IHC caption). For IF/ICC, the same SKU has a HeLa-cell IF image at 5 μg/ml (A01546-3 IF caption). Its listed reactivity covers human, mouse, rat and monkey, but the supplied IHC image documents human tissue only; clonality is unreported (catalog reactivity; A01546-3 IHC caption; 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.