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ABI2 / Abl interactor 2 · IHC design guide
Design Immunohistochemistry for ABI2
Plan ABI2 chromogenic IHC in paraffin sections using the A04302 protocol at 2 μg/ml (datasheet: A04302). Use strong neuronal and Purkinje cell staining as tissue reference points, while checking antibody specificity because HPA reports medium consistency with RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABI2 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 | Expected: cytoplasm, nucleus, projections, junctions (UniProt) | |
| Staining pattern | Strong staining in CNS neurons and Purkinje cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04302) | |
| Positive control | Adrenal gland+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 | Isoform 1, unlike isoform 3, reaches protrusion tips (UniProt) | |
| Regulation | CNS enriched; intensity varies by tissue (HPA tissue IHC) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended ABI2 IHC & IF Protocols
The catalog antibody protocol is followed by published ABI2 IHC methods for mouse femur, paired human TNBC samples, and HCC specimens (PMC12690970; PMC11212232; PMC9825985).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04302) |
| Fixation | Image fixative and duration unreported (datasheet A04302); 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 A04302); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04302) |
| Primary antibody | Rabbit anti-ABI2, 2-5 μg/ml (datasheet A04302) |
| Primary incubation | Overnight at 4 °C (datasheet A04302) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04302) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ABI2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Expression in most tissues, most abundant in CNS. No signal in the no-primary control. |
💡Decision noteFor the catalog antibody, start with heat-mediated EDTA retrieval at pH 8.0 (datasheet A04302). The cited article excerpts do not specify retrieval conditions (PMC12690970; PMC11212232; PMC9825985).
Section 2
What Is the Expected ABI2 Staining Pattern?
ABI2 is a nonmembrane protein associated with the cytoplasm, nucleus, cell projections and adherens junctions (UniProt Q9NYB9 topology and localization). In paraffin IHC, expect staining chiefly in neuronal cells, Purkinje cells and adrenal glandular cells, which HPA scores High (HPA tissue IHC). HPA describes expression across most tissues, with greatest abundance in the CNS; its tissue profile has Enhanced reliability but awaits external verification (HPA tissue IHC).
| Strong staining in cerebellar Purkinje cells, cortical neurons or adrenal glandular cells. | These are HPA High cell populations and useful positive references (HPA tissue IHC). Assess staining in the identified cells, not by whole-section darkness (general IHC practice). |
| Cell-associated cytoplasmic or nuclear staining, with possible signal near cell edges. | Cytoplasm, nucleus, projections and adherens junctions are reported locations (UniProt Q9NYB9 localization). Fine projection-tip patterns may be difficult to resolve in chromogenic sections (general IHC practice). |
| Predominant staining of a compartment inconsistent with the expected cellular pattern, such as a crisp plasma-membrane rim throughout the section. | ABI2 has no transmembrane segment (UniProt Q9NYB9 topology). Recheck morphology and controls before interpreting a pervasive membrane-rim pattern as ABI2 (general IHC practice). |
| Strong staining in adipocytes, cardiomyocytes or prostate glandular cells, especially when positive controls also look atypical. | HPA reports these cell populations as Not detected by tissue IHC (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogen activity, and inspect controls (general IHC practice). |
| Uniform haze across cells and stroma, or no staining in an HPA High reference tissue. | Haze lacks the reported cell-selective pattern (HPA tissue IHC); absent reference staining leaves the run uninterpretable. Check detection background or assay performance before scoring ABI2 (general IHC practice). |
💡Expected ABI2 appearanceCall positive when identifiable neurons, Purkinje cells or adrenal glandular cells show clear cell-associated staining, potentially cytoplasmic or nuclear, with relatively strong signal in these HPA High populations; uniform haze or dominant staining of HPA Not detected cells is suspect (HPA tissue IHC; UniProt Q9NYB9 localization; general IHC practice).
| Tissue and cell selection | HPA scores Purkinje cells, cortical neurons and adrenal glandular cells High, but breast glandular and bronchial ciliated cells Medium (HPA tissue IHC). Compare the named cell population within each tissue rather than imposing one intensity threshold across tissues. |
| Subcellular interpretation | UniProt lists cytoplasm, nucleus, cytoskeleton, projections and adherens junctions (UniProt Q9NYB9 localization). A paraffin-section pattern may emphasize a subset; judge localization alongside cell identity and the positive control (general IHC practice). |
| Isoform and resolution | Four isoforms are listed; isoform 1, unlike isoform 3, reaches protruding lamellipodia and filopodia tips (UniProt Q9NYB9 isoforms and localization). A lack of visible tip staining in IHC does not alone exclude ABI2 (general IHC practice). |
| Topology and processing | ABI2 lacks a transmembrane segment, signal peptide and propeptide; the listed chain spans residues 1–513 (UniProt Q9NYB9 topology and processing). The supplied record gives no antibody epitope, so it cannot predict epitope-specific retrieval or isoform detection. |
| Antibody evidence | The listed antibody HPA070567 has IHC Enhanced validation, meaning its pattern was reproduced by independent antibodies or orthogonal data (HPA antibodies). The tissue profile still notes medium RNA–staining consistency and pending external verification (HPA tissue IHC). |
| IF/ICC: where should ABI2 appear? | HPA reports mainly supported nucleoplasmic localization, additional supported cytosol and uncertain vesicles; the listed antibody has ICC Supported status (HPA subcellular; HPA antibodies). Use that as an IF interpretation reference, not an IHC compartment requirement. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in Purkinje cells, cortical neurons or adrenal glandular cells. | These populations are HPA High, so absent staining may reflect an unsuccessful IHC run or unsuitable assay conditions (HPA tissue IHC; general IHC practice). | Verify tissue identity and the positive control, then review the catalog antibody's IHC-P instructions for retrieval and dilution; no ABI2-specific retrieval condition is supplied here (general IHC practice). |
| A supposedly negative section shows widespread brown precipitate. | Endogenous detection activity, nonspecific antibody binding or residual reagent can produce background in chromogenic IHC (general IHC practice). | Inspect a no-primary control, review detection-system blocking and washes, and compare staining with the expected HPA cell distribution (general IHC practice; HPA tissue IHC). |
| Adipocytes, cardiomyocytes or prostate glandular cells stain more strongly than the reference cells. | HPA reports these cell populations as Not detected, while the selected neuronal and adrenal populations are High (HPA tissue IHC). | Confirm cell identity and compare controls; if the discordant pattern persists, treat specificity as unresolved rather than assigning ABI2 positivity (general IHC practice). |
| Only faint staining appears in breast or bronchus. | HPA scores breast glandular cells and bronchial ciliated cell bodies Medium, below its High reference populations (HPA tissue IHC). | Run an HPA High reference tissue alongside them and score the specified cells separately; faint signal alone does not establish assay failure (HPA tissue IHC; general IHC practice). |
| Signal appears mainly nuclear, raising concern because ABI2 regulates actin. | Nuclear localization is listed by UniProt, and HPA IF reports supported nucleoplasmic localization (UniProt Q9NYB9 localization; HPA subcellular). | Check the nuclear counterstain, cell identity and controls before rejecting nuclear signal; do not require visible projection tips in paraffin IHC (general IHC practice). |
| The positive control stains, but the test section has diffuse haze that obscures cells. | The HPA reference pattern identifies particular stained cell populations; uniform haze cannot be scored as that pattern (HPA tissue IHC; general IHC practice). | Review no-primary background, blocking, detection and washes; adjust conditions using the catalog IHC-P instructions, then rescore only clearly identifiable cells (general IHC practice). |
Experiment evidence
IHC & IF images showing ABI2 staining
A04302 · IHCIHC analysis of ABI2 using anti-ABI2 antibody (A04302). ABI2 was detected in a paraffin-embedded section of mouse brain tissue. Heat mediate
A04302 · IHCIHC analysis of ABI2 using anti-ABI2 antibody (A04302). ABI2 was detected in a paraffin-embedded section of rat brain tissue. Heat mediated
A04302 · IFIF analysis of SNF2H/SMARCA5 and Tubulin beta using anti-SNF2H/SMARCA5 antibody (A04302) and anti-Tubulin beta antibody (M05613-4). SNF2H/SM
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138443-ABI2/tissue/cerebral+cortex#imid_19139235
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138443-ABI2/tissue/colon#imid_19139145
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138443-ABI2/tissue/liver#imid_19139146
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000138443-ABI2/tissue/kidney#imid_19139163
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA070567; green = ABI2, blue = nucleus, red = microtubules.
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA070567; green = ABI2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABI2 IHC & IF
🧪Run cerebellum first and assess Purkinje cells, where ABI2 staining is High (HPA: High in Purkinje cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); neighboring unstained cells on the cerebellum slide should show counterstain without specific chromogen, but are background comparators rather than verified ABI2-negative cells.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABI2 in PC-3, SH-SY5Y, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit isotype control for the rabbit catalog antibody; and ABI2 knockout tissue or a validated immunizing-peptide block as a biological specificity control (caption: rabbit anti-ABI2 A04302; standard IHC practice). Block endogenous peroxidase and check cerebellar neuronal pigment against the controls when interpreting brown DAB signal (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04302 paraffin-section caption does not state the fixative (caption: fixative unreported). Its mouse brain example uses heat-mediated EDTA retrieval at pH 8.0, which provides a starting condition but does not establish that retrieval is required (caption: EDTA pH 8.0). Whether frozen sections or IF are easier is unreported; neuronal lipofuscin can resemble DAB pigment or autofluoresce, so compare signal with controls (standard IHC/IF practice).
HPA tissue IHC evidence for ABI2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABI2 IHC Tips
Use the A04302 paraffin-section result as a starting point, then judge ABI2 staining by compartment, cell type, controls, and section quality.
What retrieval should I try first if ABI2 staining is weak in paraffin sections?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A04302). This is the retrieval used for the catalog antibody in the mouse brain section; the caption reports paraffin embedding but does not state a fixative (caption A04302). If staining is weak, compare retrieval duration on matched sections while holding the 2 μg/ml primary concentration and overnight 4°C incubation constant (caption A04302; standard IHC practice). Judge improvement by cellular staining in the section, because ABI2 can occupy cytoplasm and nucleus, while isoform 1 can also reach projection tips (UniProt Q9NYB9 localisation).
Could fixation explain a weak or patchy ABI2 IHC result?
ABI2-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not report its fixative (caption A04302). Record fixative, fixation duration, tissue thickness, and time before fixation for each specimen, then compare sections processed together with the same EDTA pH 8.0 retrieval (standard IHC practice; datasheet A04302). Fixation can alter epitope accessibility and morphology in paraffin IHC, so vary one processing step at a time and inspect tissue preservation alongside signal (standard IHC practice). Do not attribute a weak result to ABI2 phosphorylation or its lack of a transmembrane segment without direct fixation evidence (UniProt Q9NYB9 modifications and topology).
Where should convincing ABI2 staining appear in a paraffin section?
Assess cytoplasmic and nuclear staining in intact cells, and consider staining near cell junctions or projections where morphology permits (UniProt Q9NYB9 localisation). In cell imaging, nucleoplasm is the supported main location and cytosol is also supported; vesicular localisation is uncertain (HPA subcellular). Projection-tip localisation applies to isoform 1 but not isoform 3, so its absence alone cannot exclude ABI2 in a paraffin section (UniProt Q9NYB9 isoforms and localisation). Compare cellular signal with morphology and a negative control on adjacent sections before assigning fine structures that chromogenic resolution may not separate (standard IHC practice).
Can this IHC result distinguish ABI2 isoforms or phosphorylation states?
ABI2 has 4 listed isoforms, and isoforms 1 and 3 differ in reported projection-tip localisation (UniProt Q9NYB9 isoforms and localisation). Check the antibody's stated immunogen or mapped epitope against each isoform before interpreting a missing compartment as absent protein (standard IHC practice). The record lists phosphorylation sites including serines 40, 183, and 227, but a total-ABI2 stain cannot identify those modifications without a validated site-specific reagent (UniProt Q9NYB9 modified residues; standard IHC practice). Score the observed compartment and cell population separately, and reserve isoform or phosphorylation claims for an assay that distinguishes them (standard IHC practice).
How should I plan IF follow-up to an ABI2 chromogenic IHC pattern?
Use the IHC pattern to select a cell population, then multiplex ABI2 with an independently validated marker for that expected cell type (standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue in each channel, especially when tissue autofluorescence could resemble weak ABI2 signal (standard IF practice). ABI2 has no transmembrane segment and is reported in nuclear and cytoplasmic compartments, so choose permeabilisation that gives antibody access to the compartment containing the epitope (UniProt Q9NYB9 topology and localisation; standard IF practice). Treat the paraffin-section EDTA pH 8.0 result as IHC evidence; establish IF fixation and permeabilisation empirically (datasheet A04302; standard IF practice).
How can I reduce diffuse DAB or nonspecific ABI2 staining?
First compare the test section with a no-primary control and inspect whether brown deposit follows cells, tissue edges, or damaged regions (standard IHC practice). The reported mouse brain IHC used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and an HRP/DAB detection system (caption A04302). Apply a peroxidase block before HRP detection, maintain consistent washes, and titrate primary concentration or DAB development if the control or tissue shows excess deposit (standard IHC practice). Retain plausible nuclear or cytoplasmic signal while assessing background, since both compartments are reported for ABI2 (UniProt Q9NYB9 localisation).
What should I score when comparing ABI2 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since ABI2 can be cytoplasmic or nuclear and HPA reports different staining levels across cell types (UniProt Q9NYB9 localisation; HPA tissue IHC). For intact target cells, record the 0–3 intensity category and percentage in each category to calculate an H-score, or report the percentage positive using one fixed threshold (standard IHC practice). For spatial comparisons, count positive cells per mm² of viable tissue rather than per whole image, and exclude folds and necrosis (standard IHC practice). Normalise comparisons to the same cell population, tissue area, staining batch, and image settings, with negative controls reviewed alongside (standard IHC practice).
How do I distinguish genuine ABI2 staining from an artefact?
Give greatest weight to staining in intact cells with plausible cytoplasmic or nuclear distribution, since both are reported for ABI2 (UniProt Q9NYB9 localisation). HPA reports high staining in cerebellar Purkinje cells and cerebral-cortex neuronal cells, while adipocytes were not detected; use those observations as context, not as universal controls (HPA tissue IHC). Suspect artefact when signal is confined to section edges, folds, or necrotic areas, or appears in the no-primary control after HRP/DAB development (standard IHC practice). The HPA tissue assessment is Enhanced, with medium staining–RNA consistency and external verification pending, so resolve surprising cell patterns with independent evidence (HPA tissue IHC).
Boster reagents
Best ABI2 / Abl interactor 2 IHC Antibodies
Real ABI2 IHC images show paraffin sections of human, mouse, and rat brain (A04302 IHC captions). IF/ICC is listed, but the supplied IF caption identifies a different target (catalog IF caption).
Real IHC data
Anti-ABI2 Antibody ®
Cat # A04302
A04302 is the only SKU with a rendered card and its own IHC figure, showing ABI2 staining in a mouse brain paraffin section (A04302 card caption). Additional A04302 IHC captions describe rat and human brain paraffin sections; its applications also list IF/ICC (A04302 IHC captions; catalog applications).
Which to pick: Choose A04302 for paraffin-section IHC: its own brain captions document staining across human, mouse, and rat, with 2 μg/ml primary antibody after EDTA pH 8.0 retrieval; the fixative is unreported (A04302 IHC captions). For IF/ICC, PA2068 lists both applications and 5 μg/ml for human immunocytochemistry/immunofluorescence, but has no supplied IF image (PA2068 catalog applications and dilution; IF image alts). For cross-species IHC, A04302 has the strongest pictured support across the three species; M04302 lists IHC and human, mouse, and rat reactivity but has no supplied IHC figure (A04302 IHC captions; M04302 catalog applications, reactivity, and IHC image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NYB9 (ABI2_HUMAN, Abl interactor 2).
- Human Protein Atlas. ABI2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ABI2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to vesicles and cytosol..
- Human Protein Atlas. ABI2 antibody validation summary (1 antibodies).
- Arsenic Sulfide Induces Apoptosis in Myelodysplastic Neoplasm Cells Through the Suppression of ABI2. Biological procedures online 2025 — PMC12690970.
- Inhibition of ABI2 ubiquitination-dependent degradation suppresses TNBC cell growth via down-regulating PI3K/Akt signaling pathway. Cancer cell international 2024 — PMC11212232.
- ABI2-mediated MEOX2/KLF4-NANOG axis promotes liver cancer stem cell and drives tumour recurrence. Liver international : official journal of the International Association for the Study of the Liver 2022 — PMC9825985.
- PIM1 phosphorylates ABI2 to enhance actin dynamics and promote tumor invasion. The Journal of cell biology 2023 — PMC10103708.
- PubMed PMID:7590236 — UniProt-cited evidence.
- PubMed PMID:8649853 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABI2 (Abl interactor 2). Expected localisation Expected: cytoplasm, nucleus, projections, junctions (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04302). Validated IHC antibody: A04302 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A04302); 2. Fixation: Image fixative and duration unreported (datasheet A04302); 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 retrieval: EDTA pH 8.0 (datasheet A04302); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04302); 8. Primary antibody: Rabbit anti-ABI2, 2-5 μg/ml (datasheet A04302); 9. Primary incubation: Overnight at 4 °C (datasheet A04302). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Isoform 1, unlike isoform 3, reaches protrusion tips (UniProt); Isoform / epitope: 4 isoforms; epitope coverage is unspecified (UniProt); Regulation: CNS enriched; intensity varies by tissue (HPA tissue IHC). Evidence: UniProt Q9NYB9, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to…, Datasheet A04302, PMC12690970, PMC11212232. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABI2/immunohistochemistry-ihc-guide.