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ABI2 / Abl interactor 2 · IHC design guide
Design Immunohistochemistry for ABI2
Plan ABI2 chromogenic IHC on paraffin sections using cerebellar Purkinje cells as a positive reference and adipocytes as a negative comparator (HPA tissue IHC). The A04302 tissue-IHC caption uses 2 μg/ml primary antibody overnight at 4°C, followed by peroxidase and DAB detection (datasheet A04302).
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 | Cytoplasm and nucleus expected; tissue compartment unreported (UniProt; HPA tissue IHC) | |
| Staining pattern | High in Purkinje and cortical neurons and adrenal glandular 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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining has only medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | No expression regulator specified (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage unspecified (UniProt) |
Section 1
Recommended ABI2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A04302) is accompanied by ABI2 IHC methods from three articles (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 noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet: A04302); the mouse femur method reports retrieval without conditions (PMC12690970).
Section 2
What Is the Expected ABI2 Staining Pattern?
ABI2 can localize to the nucleus, cytoplasm, actin-rich projections and adherens junctions; it has no transmembrane segment (UniProt Q9NYB9). In paraffin-section IHC, expect staining chiefly in neuronal cells of the CNS, with high staining also reported in adrenal glandular cells (HPA tissue IHC). HPA rates the tissue profile Enhanced, while noting medium consistency with RNA data and pending external verification (HPA tissue IHC).
| Neuronal cells in cerebral cortex or Purkinje cells in cerebellum stain strongly, with intracellular signal (HPA tissue IHC; UniProt Q9NYB9). | This fits HPA's High staining in those cell types and UniProt's nuclear and cytoplasmic localization. Assess the named cells rather than treating uniform staining of every structure in the section as the expected pattern (HPA tissue IHC; UniProt Q9NYB9; general IHC practice). |
| Signal is concentrated in nuclei, with some cytoplasmic staining (HPA subcellular ICC-IF; UniProt Q9NYB9). | Nucleoplasmic localization is supported by HPA ICC-IF, and nuclear plus cytoplasmic localization is listed by UniProt. ICC-IF compartment evidence can guide interpretation, but it does not establish the exact appearance of chromogenic paraffin-section IHC (HPA subcellular ICC-IF; UniProt Q9NYB9). |
| Staining appears only in extracellular deposits, lumens or a sharp continuous membrane outline (UniProt Q9NYB9 topology). | Treat an exclusively extracellular or membrane-bound pattern as suspect: ABI2 has no transmembrane segment, while its listed locations are intracellular or associated with cell projections and junctions. Check section morphology and detection controls before assigning it to ABI2 (UniProt Q9NYB9; general IHC practice). |
| Strong signal appears in adipocytes or cardiomyocytes (HPA tissue IHC: Not detected). | HPA reports ABI2 as Not detected in these cells. Unexpected staining may reflect cross-reactivity or endogenous detection activity; compare a reagent-control section. An HPA non-detection is a reference observation, not proof that every specimen must be negative (HPA tissue IHC; general IHC practice). |
| Color spreads across the section without a clear cellular pattern, or no signal appears in a known-positive region (HPA tissue IHC). | Widespread haze is background rather than a scorable localization pattern; absent signal in HPA High neuronal or adrenal glandular cells makes that run hard to interpret. Review controls and the staining workflow before drawing a biological conclusion (HPA tissue IHC; general IHC practice). |
💡Expected ABI2 appearanceCall a result consistent with ABI2 when intracellular staining is clear and relatively strong in cortical neurons, cerebellar Purkinje cells or adrenal glandular cells; diffuse noncellular color or staining confined to extracellular space is suspect (HPA tissue IHC: High in those cells; UniProt Q9NYB9 localization; general IHC practice).
| Choice of positive and reference-negative cells | HPA reports High staining in cortical neurons, Purkinje cells and adrenal glandular cells, but Not detected in adipocytes, cardiomyocytes and several other listed cell types. Use the specified cell population, not an entire organ, for comparison (HPA tissue IHC). |
| Compartment and fine structure | UniProt lists nucleus, cytoplasm, cytoskeleton, lamellipodia, filopodia and nascent adherens junctions. Fine projection or junction detail may be difficult to resolve in routine chromogenic sections; score what is visibly cellular (UniProt Q9NYB9; general IHC practice). |
| Isoform-dependent localization | UniProt lists four ABI2 isoforms and reports protruding lamellipodia and filopodia-tip localization for isoform 1 but not isoform 3. Lack of tip staining alone therefore cannot exclude ABI2; no isoform-specific behavior for the catalog antibody is supplied (UniProt Q9NYB9). |
| Antibody evidence and limits | The supplied HPA antibody record lists HPA070567 as IHC Enhanced and ICC Supported. HPA's tissue-level Enhanced profile remains pending external verification, so resolve discordant staining with controls and the named cell-level references (HPA antibody record; HPA tissue IHC). |
| Q: What does ICC-IF show? | A: Mainly nucleoplasmic ABI2, with supported cytosolic and uncertain vesicular localization in the supplied HPA subcellular record. Those are ICC-IF observations; this section's primary interpretation concerns chromogenic paraffin-section IHC (HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in cortical neurons or cerebellar Purkinje cells (HPA tissue IHC: High). | The run may have failed at retrieval, antibody incubation or detection; the observation alone does not identify which step failed (general IHC practice). | Confirm the expected cells are present, then review the catalog antibody's IHC-P instructions and run controls before changing one workflow variable at a time (HPA tissue IHC; general IHC practice). |
| Only diffuse color is visible across the section (general IHC practice). | Background from nonspecific binding or detection chemistry can obscure a cell-level pattern (general IHC practice). | Compare a section without primary antibody, inspect blocking and washes, and reassess antibody dilution against the catalog IHC-P instructions; no ABI2-specific dilution is supplied here (general IHC practice). |
| Adipocytes or cardiomyocytes stain strongly (HPA tissue IHC: Not detected). | Cross-reactivity or endogenous detection activity is possible; HPA non-detection alone cannot identify the cause (HPA tissue IHC; general IHC practice). | Check no-primary and detection-reagent controls, then compare the cellular pattern with an HPA High reference region in the same run (HPA tissue IHC; general IHC practice). |
| Staining is confined to an extracellular or lumenal structure (UniProt Q9NYB9 topology). | That pattern conflicts with the supplied intracellular ABI2 locations and lack of a transmembrane segment (UniProt Q9NYB9). | Inspect morphology and reagent controls; require convincing intracellular staining in an HPA High cell population before calling the structure ABI2-positive (UniProt Q9NYB9; HPA tissue IHC; general IHC practice). |
| Nuclear staining is called an artifact solely because cytoplasmic staining was expected (HPA subcellular ICC-IF). | The expectation is too narrow: HPA supports nucleoplasmic localization, and UniProt lists both nucleus and cytoplasm (HPA subcellular ICC-IF; UniProt Q9NYB9). | Reassess which cells stain and whether the signal is confined to nuclei or accompanies cytoplasmic staining; use section controls to judge specificity (HPA tissue IHC; HPA subcellular ICC-IF; general IHC practice). |
| One specimen disagrees with the HPA tissue profile (HPA tissue IHC). | HPA notes medium staining-to-RNA consistency and pending external verification; a single discrepancy does not establish the reason (HPA tissue IHC). | Repeat with appropriate run controls and score the named cell population. Record the discrepancy without treating an HPA High or Not detected category as an absolute rule for every specimen (HPA tissue IHC; 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 require staining in Purkinje cells (HPA: High in Purkinje cells). Use adipose tissue as the negative comparator, scoring adipocytes (HPA: Not detected in adipocytes); on the cerebellum slide, treat non-Purkinje cells as internal negatives only if they remain at background intensity in the no-primary comparison, since their ABI2 status is not specified by the supplied HPA row.
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) slide, a concentration-matched rabbit IgG isotype control, and, if available, ABI2 knockout tissue or antigen-peptide competition (caption: rabbit anti-ABI2; standard IHC practice). Quench endogenous peroxidase and assess background in the cerebellum before interpreting DAB staining (caption: peroxidase secondary and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected A04302 paraffin-section caption is unreported (selected tissue-IHC caption). That caption uses heat-mediated retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; in brain sections, endogenous peroxidase or pigment can complicate chromogenic scoring (standard IHC 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 catalog antibody’s paraffin-section protocol as the starting point, then assess staining against ABI2’s documented cellular distribution and appropriate controls.
How should I adjust retrieval when ABI2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A04302). The selected mouse-brain IHC example used that retrieval before 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (caption A04302). If staining is weak, vary heating duration within the retrieval system’s validated range while keeping section thickness, detection chemistry and exposure to DAB constant (standard IHC practice). Compare each condition with a no-primary control and assess cellular detail as well as signal, because excessive retrieval can damage morphology and increase nonspecific staining (standard IHC practice).
Could fixation explain weak or uneven ABI2 staining?
Target-specific sensitivity of ABI2 to fixation is unknown: the selected paraffin-section caption does not state a fixative (caption A04302). Record the fixative, fixation duration and tissue-processing history for every section before comparing staining intensity across specimens (standard IHC practice). If weak staining tracks a processing batch, repeat the catalog retrieval condition, EDTA at pH 8.0, on a well-preserved control section processed alongside the test tissue (datasheet A04302; standard IHC practice). Interpret any improvement as evidence about that processing workflow, not proof of an ABI2-specific fixation effect, and evaluate morphology before scoring signal (standard IHC practice).
Which cellular compartments should count as plausible ABI2 staining?
Assess cytoplasmic and nuclear staining as potentially plausible, since both locations are recorded for ABI2 (UniProt Q9NYB9). Nucleoplasmic and cytosolic localisation have supporting cell-imaging evidence, whereas the reported vesicle location is uncertain (HPA subcellular). ABI2 also occurs at actin-rich projections and nascent adherens junctions; isoform 1, unlike isoform 3, reaches protruding lamellipodia and filopodia tips (UniProt Q9NYB9). In chromogenic sections, score these fine structures only when morphology resolves them clearly, and document nuclear and cytoplasmic signal separately so a compartment shift does not masquerade as a change in total expression (standard IHC practice).
Can this stain distinguish ABI2 isoforms or modified epitopes?
ABI2 has 4 annotated isoforms, and isoforms 1 and 3 differ in their reported localisation to protruding lamellipodia and filopodia tips (UniProt Q9NYB9). The supplied caption identifies the catalog antibody and IHC conditions but gives no epitope sequence or isoform-reactivity data (caption A04302). Therefore, treat a positive paraffin-section stain as ABI2-associated signal without assigning it to a particular isoform or phosphorylation state (standard IHC interpretation). ABI2 has annotated phosphorylation sites and an SH3 domain at residues 451–510; establish any epitope-specific interpretation with documented antibody mapping and an appropriate orthogonal assay (UniProt Q9NYB9; standard IHC practice).
How can IF help resolve ABI2 staining patterns seen by chromogenic IHC?
For a separate IF experiment, multiplex ABI2 with a neuronal-cell marker when examining the CNS populations highlighted by tissue IHC (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red ABI2 channel if the specimen shows strong autofluorescence in shorter-wavelength channels (standard IF practice). ABI2 has no transmembrane segment and is reported in intracellular compartments, so permeabilisation must allow antibody access to cytosolic or nuclear epitopes (UniProt Q9NYB9; standard IF practice). Compare single-stain and no-primary controls, then use the compartment-resolved IF pattern to investigate ambiguous DAB signal without assuming that IF and paraffin-section IHC produce identical localisation (standard IF practice).
What should I change if ABI2 DAB staining is widespread and diffuse?
Begin with a no-primary section to separate primary-antibody-dependent staining from detection-system background (standard IHC practice). The selected IHC example used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary antibody (caption A04302). Check whether endogenous peroxidase was adequately blocked, then adjust primary concentration, washing and DAB development one variable at a time while preserving a positive-control section (standard IHC practice). Diffuse colour should be weighed against ABI2’s reported nuclear, cytoplasmic and projection-associated locations and against preserved cell boundaries, rather than accepted solely because the tissue looks dark (UniProt Q9NYB9; standard IHC practice).
How should I quantify ABI2 across paraffin-section samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, using the same section-selection rule for every specimen (standard IHC practice). Record the percentage of positive cells and staining intensity as an H-score, or count positive cells per mm² when cell density is the main outcome (standard IHC practice). Normalise cell counts to the measured tissue area or eligible-cell count, and keep retrieval, antibody concentration, DAB development and image settings consistent across batches (standard IHC practice). Score nuclear and cytoplasmic compartments separately if relevant, and report excluded damaged or necrotic areas so changes in morphology do not distort ABI2 comparisons (UniProt Q9NYB9; standard IHC practice).
How do I distinguish convincing ABI2 staining from artefact?
Look for reproducible cellular staining in plausible compartments, including nucleus and cytoplasm, rather than colour restricted to torn edges or necrotic regions (UniProt Q9NYB9; standard IHC practice). CNS neuronal populations offer a tissue-IHC reference, including high staining in cerebellar Purkinje cells and cerebral-cortex neuronal cells (HPA tissue IHC). Investigate strong staining in a cell population reported as undetected by HPA, such as adipocytes, with morphology and controls before calling it ABI2-positive (HPA tissue IHC; standard IHC practice). A no-primary section and adequate endogenous-peroxidase block help identify detection artefacts; HPA’s enhanced rating still carries a medium RNA-to-staining consistency caveat (standard IHC practice; HPA tissue IHC).
Boster reagents
Best ABI2 / Abl interactor 2 IHC Antibodies
A04302 has ABI2 IHC images from paraffin sections of mouse, rat, and human brain (A04302 IHC captions). IF/ICC is listed as an application, but its IF caption identifies another target (A04302 catalog).
Real IHC data
Anti-ABI2 Antibody ®
Cat # A04302
A04302 is the SKU that will render; its IHC captions show staining in paraffin sections of mouse, rat, and human brain (A04302 IHC captions). Its catalog lists IF/ICC, but the supplied IF caption identifies SNF2H/SMARCA5 rather than ABI2, so it does not establish ABI2 IF staining (A04302 catalog; A04302 IF caption).
Which to pick: Choose A04302 for tissue IHC because its own captions document paraffin-section staining; the fixative is unreported (A04302 IHC captions). For IF/ICC, PA2068 lists that application, but the payload supplies no IF image for it (PA2068 catalog). For work across human, mouse, and rat, A04302 lists reactivity with all three and has an IHC caption for each species (A04302 catalog; A04302 IHC captions).
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 Cytoplasm and nucleus expected; tissue compartment unreported (UniProt; HPA tissue IHC). 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 paraffin sections (standard IHC practice; not target-specific); Caveat: Tissue staining has only medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 4 isoforms; epitope coverage unspecified (UniProt); Regulation: No expression regulator specified (UniProt). 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.