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ABI1 / Abl interactor 1 · IHC design guide
Design Immunohistochemistry for ABI1
Plan ABI1 chromogenic IHC in paraffin sections using cytoplasmic staining as the tissue readout (HPA tissue IHC). The guide identifies positive control tissues (HPA tissue IHC) and a catalog antibody's IHC-P starting concentration of 2–5 μg/mL (datasheet PA1001).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ABI1 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 | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9416) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Smooth muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Smooth muscle cells have no detected staining (HPA tissue IHC) | |
| Regulation | No specific inducer reported (UniProt) | |
| Isoform / epitope | 12 isoforms; epitope coverage unspecified (UniProt; catalog datasheets) |
Section 1
Recommended ABI1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet PB9416). The published options cover colonic lesions (PMC3393686) and ovarian cancer sections (PMC3059077).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet PB9416) |
| Fixation | Image fixative and duration unreported (datasheet PB9416); 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 PB9416); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9416) |
| Primary antibody | Rabbit anti-ABI1, 2-5μg/ml (datasheet PB9416) |
| Primary incubation | Overnight at 4 °C (datasheet PB9416) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9416) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ABI1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet PB9416); neither published excerpt specifies a retrieval method (PMC3393686; PMC3059077).
Section 2
What Is the Expected ABI1 Staining Pattern?
ABI1 is a non-transmembrane protein found in the cytoplasm, nucleus, cell projections and neuronal synaptic regions (UniProt Q8IZP0 localization and topology). In paraffin-section IHC, expect mainly cytoplasmic staining across many cell types, with high staining reported in selected glandular, hematopoietic, respiratory epithelial and neuronal cells (HPA tissue IHC). HPA rates the tissue pattern Approved, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Cytoplasmic staining in appendix or colon glandular cells, or in cerebral cortex neurons. | This fits cells scored High by HPA and its ubiquitous cytoplasmic IHC profile (HPA tissue IHC). Score the named cell population and compartment separately; a tissue label alone does not mean every cell should stain. Use the expected pattern as a comparison, while retaining HPA's medium-consistency and pending-verification caveat (HPA tissue IHC). |
| Predominantly nuclear-only staining, or a sharp surface outline with little cytoplasmic signal. | Neither is the reported dominant tissue-IHC pattern (HPA tissue IHC). Nuclear localization is biologically possible, so nuclear signal alone does not prove an artefact (UniProt Q8IZP0 localization). Check morphology and controls before interpreting an unusual pattern; the membrane and junction pattern reported by ICC-IF does not establish the expected chromogenic IHC pattern (HPA subcellular ICC-IF). |
| Strong staining of smooth muscle cells while expected positive cells also stain. | Smooth muscle cells are reported Not detected, making this a discordant cell-type pattern (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity, then compare antibody-omission and detection controls (general IHC practice). Do not treat adjacent positive cells as evidence that smooth muscle staining is ABI1-specific. |
| Color spreads across stroma, lumens or most cells without clear cellular boundaries. | A diffuse field is difficult to reconcile with interpretable cytoplasmic staining in identifiable cells (HPA tissue IHC). Review the reagent-omission control, blocking and detection steps for nonspecific or endogenous color development (general IHC practice). Score only signal that can be assigned to cells and a compartment. |
| No staining in a listed high-staining cell population. | Absence in, for example, appendix glandular cells conflicts with HPA's High observation (HPA tissue IHC). First confirm that the expected cells are present and the detection control worked (general IHC practice). A negative slide alone cannot distinguish a technical failure from sample variation or the limits of HPA's Approved, medium-consistency evidence (HPA tissue IHC). |
💡Expected ABI1 appearanceCall a convincing positive result cell-associated, predominantly cytoplasmic chromogenic staining in an HPA High population, with distinguishable cell boundaries; strong smooth muscle staining or diffuse color without cell boundaries needs control review (HPA tissue IHC; general IHC practice).
| Which cells provide a comparison? | HPA reports High staining in appendix, breast, cervix and colon glandular cells; bone marrow hematopoietic cells; bronchial respiratory epithelium; and selected cerebellar and cortical cells. Smooth muscle cells are Not detected (HPA tissue IHC). |
| Does IF/ICC require the same pattern? | No. ICC-IF reports approved plasma membrane and cell-junction localization, whereas tissue IHC reports a ubiquitous cytoplasmic profile (HPA subcellular ICC-IF; HPA tissue IHC). Interpret each assay against its own observed pattern. |
| How firm is antibody support? | The listed mouse monoclonal CAB008375 is Approved for IHC; rabbit polyclonal HPA068407 is Approved for ICC, with no IHC status listed (HPA antibody records). HPA describes tissue-IHC agreement as medium, pending external verification (HPA tissue IHC). |
| Could isoforms or epitope choice matter? | UniProt lists 12 ABI1 isoforms and an SH3 domain at residues 446–505 (UniProt Q8IZP0). The supplied record gives no IHC antibody epitope, so it cannot establish which isoforms the IHC signal represents. |
| Does antigen retrieval have a specified ABI1 effect? | Retrieval is a standard paraffin-IHC variable (general IHC practice). The supplied HPA and UniProt records give no ABI1-specific retrieval condition or fixation-sensitivity result; judge a chosen condition by tissue morphology and controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected High cells are unstained. | The cell population may be absent from the section, or staining or detection may have failed (general IHC practice). | Locate the named population first, then check a known-positive section and the detection control before changing conditions. Appendix or colon glandular cells are reported High (HPA tissue IHC). |
| Smooth muscle cells stain strongly. | This conflicts with the Not detected HPA observation; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Check antibody-omission and detection controls, then reassess whether color is truly within smooth muscle cells (general IHC practice). |
| Signal is uniformly nuclear. | Nuclear ABI1 is possible, but a nuclear-only field differs from HPA's predominant cytoplasmic tissue-IHC profile (UniProt Q8IZP0 localization; HPA tissue IHC). | Confirm cell boundaries and compare a known-positive section and omission control before assigning the nuclear signal to ABI1 (general IHC practice). |
| Background obscures cell boundaries. | Nonspecific reagent binding or endogenous detection activity can produce diffuse chromogenic color (general IHC practice). | Inspect omission controls and blocking and detection steps; interpret only cell-associated signal with a readable compartment (general IHC practice). |
| Staining differs across tissues. | HPA reports both High and Low cell populations, despite an overall ubiquitous cytoplasmic profile (HPA tissue IHC). | Compare the same named cell types and compartments. Record intensity by cell population, rather than treating every tissue as an equally strong positive (HPA tissue IHC). |
| An IF image looks junctional while IHC looks cytoplasmic. | Those are the respective reported ICC-IF and tissue-IHC observations (HPA subcellular ICC-IF; HPA tissue IHC). | Assess the paraffin section against the tissue-IHC profile and its controls; use the assay-specific IF/ICC guide for IF interpretation (HPA tissue IHC; HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing ABI1 staining
PB9416 · IHCIHC analysis of SSH3BP1/ABI1 using anti-SSH3BP1/ABI1 antibody (PB9416). SSH3BP1/ABI1 was detected in a paraffin-embedded section of mouse br
PB9416 · IHCIHC analysis of SSH3BP1/ABI1 using anti-SSH3BP1/ABI1 antibody (PB9416). SSH3BP1/ABI1 was detected in a paraffin-embedded section of rat brai
PB9416 · IFIF analysis of SSH3BP1/ABI1 using anti-SSH3BP1/ABI1 antibody (PB9416). SSH3BP1/ABI1 was detected in an immunocytochemical section of A431 ce
PA1001 · IHCIHC analysis of ABI1 using anti-ABI1 antibody (PA1001). ABI1 was detected in a paraffin-embedded section of human lung cancer tissue. Heat m
PA1001 · IHCIHC analysis of ABI1 using anti-ABI1 antibody (PA1001). ABI1 was detected in a paraffin-embedded section of human colorectal adenocarcinoma
PA1001 · IFIF analysis of ABI1 using anti-ABI1 antibody (PA1001). ABI1 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retr
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136754-ABI1/tissue/cerebral+cortex#imid_2243808
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136754-ABI1/tissue/colon#imid_2243734
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136754-ABI1/tissue/liver#imid_2243743
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136754-ABI1/tissue/kidney#imid_2243744
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA068407; green = ABI1, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA068407; green = ABI1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ABI1 IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). Use smooth muscle as the negative tissue (HPA: Not detected in smooth muscle cells); any cells used as internal negatives on the marrow slide should show counterstain without specific DAB signal, and their negative status must be confirmed on that slide.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Smooth muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ABI1 in CACO-2, SK-MEL-30, U2OS, with annotated localisation: Plasma membrane (approved), Cell Junctions (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), host- and clonality-matched rabbit IgG isotype, and ABI1-knockout tissue controls (caption: rabbit anti-ABI1 antibody; standard IHC controls). Quench endogenous peroxidase before HRP/DAB detection because hematopoietic cells can produce background signal (HPA: bone marrow hematopoietic cells; caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: The selected PB9416 paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but its fixative is unreported; no ABI1-specific fixation window or fixation effect is reported in the supplied evidence (caption: EDTA retrieval; caption: fixative not stated). Frozen sections or IF cannot be judged easier from these data; the HPA IF images document plasma-membrane and cell-junction localization in cell lines, not marrow tissue (HPA: subcellular ICC-IF). In marrow, endogenous peroxidase can mimic chromogenic staining, so interpret signal against the peroxidase-quenched controls (HPA: bone marrow hematopoietic cells; caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for ABI1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ABI1 IHC Tips
Troubleshoot ABI1 staining in paraffin sections by checking retrieval, cellular distribution, controls, and scoring before interpreting chromogenic signal.
Which retrieval condition should I start with for ABI1 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for the catalog antibody in paraffin sections (datasheet PB9416). The illustrated mouse brain section used that condition before overnight incubation with 2 μg/ml primary antibody at 4°C (datasheet PB9416). If staining is weak, compare retrieval durations on adjacent sections while keeping antibody concentration and DAB development constant, then inspect tissue structure and background (standard IHC practice). Record the heating and cooling conditions for every run so a brighter signal can be distinguished from increased nonspecific staining (standard IHC practice).
Could fixation explain weak ABI1 staining in my sections?
Target-specific sensitivity of ABI1 staining to fixation is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (datasheet PB9416). Record the specimen's actual fixative, fixation duration, processing history, and storage conditions before comparing staining between blocks (standard IHC practice). Run sections from differently processed blocks with the same EDTA pH 8.0 retrieval and antibody conditions, including a positive reference section in each run (datasheet PB9416; standard IHC practice). If signal tracks processing history, describe that association without assigning a molecular fixation mechanism to ABI1 (standard IHC practice).
Where should convincing ABI1 staining appear?
Assess cellular patterns rather than requiring one compartment: ABI1 is reported in cytoplasm, nucleus, cytoskeleton, and cell projections (UniProt Q8IZP0), while tissue IHC shows ubiquitous cytoplasmic expression (HPA tissue IHC). In the supplied subcellular imaging data, plasma membrane and cell junctions are approved locations (HPA subcellular). Compare staining within intact cells and across adjacent anatomical regions, using a counterstain to identify nuclei and tissue boundaries (standard IHC practice). A nuclear, junctional, or projection-associated pattern warrants comparison with cytoplasmic staining and controls before being scored as ABI1-specific (UniProt Q8IZP0; HPA tissue IHC; standard IHC practice).
Why might an ABI1 antibody miss signal in some cells?
ABI1 has 12 listed isoforms, and the supplied caption does not identify the epitope recognized by PB9416 (UniProt Q8IZP0; datasheet PB9416). Its coiled-coil homology region spans residues 45–107, and its SH3 domain spans 446–505; multiple modified residues are also listed (UniProt Q8IZP0). Check the antibody's documented immunogen or epitope before attributing regional staining differences to isoform expression or epitope masking (standard IHC practice). Compare the same section-processing conditions across samples and report unresolved epitope coverage as a limitation of the interpretation (standard IHC practice).
How should I cross-check an ABI1 IHC pattern by IF?
Use IF as a separate validation experiment and multiplex ABI1 with a marker for the cell type expected in the tissue being examined (standard IF practice). Choose spectrally separated fluorophores and image an unstained section to assess tissue autofluorescence before interpreting a weak ABI1 channel (standard IF practice). ABI1 has no transmembrane segment, but its reported locations include cytoplasm, nucleus, plasma membrane, and junctions; select permeabilisation to provide access to the antibody's documented epitope (UniProt Q8IZP0; HPA subcellular; standard IF practice). Compare the IF distribution with the chromogenic section without assuming that an IF condition validates the IHC antibody or its retrieval (standard IHC/IF practice).
How can I reduce diffuse or granular ABI1 background?
First compare the stained section with a no-primary control and inspect whether background follows tissue edges, damaged regions, or the entire slide (standard IHC practice). The illustrated PB9416 workflow used 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet PB9416). For HRP and DAB detection, include a peroxidase block and control DAB development consistently across sections; these are general chromogenic IHC steps (standard IHC practice). If background persists, titrate primary antibody and secondary detection separately while checking that expected cellular detail remains visible (standard IHC practice).
What should I measure when comparing ABI1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell population before scoring, then use the same imaging and DAB development settings for all compared sections (standard IHC practice). For cell-associated staining, report the percentage of positive cells and an intensity-weighted H-score; for selected cell populations, density per mm² may be useful (standard IHC practice). Normalize counts to the number of evaluable cells or measured tissue area, and exclude folds, necrosis, and poorly preserved regions by preset criteria (standard IHC practice). Score cytoplasmic and other compartments separately when relevant, because ABI1 has several reported locations (UniProt Q8IZP0; HPA tissue IHC; HPA subcellular).
How do I distinguish true ABI1 signal from artefact?
Compare cellular staining with a no-primary control, intact tissue morphology, and the expected compartment before calling a chromogenic signal positive (standard IHC practice). Cytoplasmic tissue staining is broadly reported, including high staining in cerebral cortex neuronal cells, whereas smooth muscle cells were not detected in the supplied tissue profile (HPA tissue IHC). Flag staining restricted to cut edges, necrotic areas, or endogenous peroxidase-rich regions for control review rather than scoring it automatically (standard IHC practice). A pattern that conflicts with the annotated cytoplasmic, nuclear, projection, membrane, or junctional locations needs independent validation before a biological explanation (UniProt Q8IZP0; HPA subcellular; standard IHC practice).
Boster reagents
Best ABI1 / Abl interactor 1 IHC Antibodies
ABI1 IHC and IF examples cover human cancer tissues, mouse and rat brain sections, and U2OS and A431 cells (catalog image captions).
Real IHC data
Anti-SSH3BP1/ABI1 Antibody ®
Cat # PB9416
Real IHC data
Anti-SSH3BP1/ABI1 Antibody ®
Cat # PA1001
PA1001 has paraffin-section IHC examples in human lung cancer and colorectal adenocarcinoma, plus IF examples in U2OS cells and human intestine and breast cancer sections (PA1001 image captions). PB9416 has paraffin-section IHC examples in mouse and rat brain and human ovarian cancer, plus an IF/ICC example in A431 cells (PB9416 image captions).
Which to pick: For human tissue IHC, choose PA1001 when its lung or colorectal examples fit your study; its paraffin-section captions report EDTA pH 8.0 retrieval and 2 μg/ml primary antibody (PA1001 IHC captions). For IF/ICC, PA1001 has U2OS-cell and tissue IF examples, while PB9416 has an A431-cell IF/ICC example; both list IF and ICC applications (catalog applications; IF image captions). For cross-species tissue IHC, PB9416 has direct mouse, rat and human paraffin-section examples; its captions report EDTA pH 8.0 retrieval and 2 μg/ml primary antibody, while neither SKU’s IHC captions report the fixative (PB9416 and PA1001 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8IZP0 (ABI1_HUMAN, Abl interactor 1).
- Human Protein Atlas. ABI1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ABI1 subcellular location (ICC-IF): Localized to the plasma membrane and cell junctions..
- Human Protein Atlas. ABI1 antibody validation summary (2 antibodies).
- Expression of Abelson interactor 1 (Abi1) correlates with inflammation, KRAS mutation and adenomatous change during colonic carcinogenesis. PloS one 2012 — PMC3393686.
- Expression and Y435-phosphorylation of Abelson interactor 1 (Abi1) promotes tumour cell adhesion, extracellular matrix degradation and invasion by colorectal carcinoma cells. Molecular cancer 2014 — PMC4066275.
- Integrity of SOS1/EPS8/ABI1 tri-complex determines ovarian cancer metastasis. Cancer research 2010 — PMC3059077.
- Fat3 acts through independent cytoskeletal effectors to coordinate asymmetric cell behaviors during polarized circuit assembly. Cell reports 2022 — PMC8865054.
- PubMed PMID:9010225 — UniProt-cited evidence.
- PubMed PMID:9593709 — UniProt-cited evidence.
- PubMed PMID:11418237 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ABI1 (Abl interactor 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9416). Validated IHC antibody: PB9416 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Smooth muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet PB9416); 2. Fixation: Image fixative and duration unreported (datasheet PB9416); 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 PB9416); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9416); 8. Primary antibody: Rabbit anti-ABI1, 2-5μg/ml (datasheet PB9416); 9. Primary incubation: Overnight at 4 °C (datasheet PB9416). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Smooth muscle cells have no detected staining (HPA tissue IHC); Isoform / epitope: 12 isoforms; epitope coverage unspecified (UniProt; catalog datasheets); Regulation: No specific inducer reported (UniProt). Evidence: UniProt Q8IZP0, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane and cell junctions., Datasheet PB9416, PMC3393686, PMC3059077. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ABI1/immunohistochemistry-ihc-guide.