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BIN1 / Myc box-dependent-interacting protein 1 · IHC design guide
Design Immunohistochemistry for BIN1
Plan BIN1 staining in paraffin sections using cytoplasmic CNS and skeletal muscle patterns as reference points (HPA tissue IHC). Compare staining with matched controls, and interpret unexpected signal cautiously because presumed off-target binding has been observed (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BIN1 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 | Predominantly cytoplasmic in CNS and skeletal muscle (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic CNS glia and myocytes; granular cerebellar cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01551-1) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Esophagus+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01551-1) | |
| Caveat | Presumed off-target binding may complicate scoring (HPA tissue IHC) | |
| Regulation | Highest expression in brain and muscle (UniProt) | |
| Isoform / epitope | 11 isoforms; epitope coverage needs checking (UniProt) |
Section 1
Recommended BIN1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A01551-1); one published BIN1 IHC protocol uses FFPE HCC tissue (PMC10495885).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A01551-1) |
| Fixation | Image fixative and duration unreported (datasheet A01551-1); 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 A01551-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01551-1) |
| Primary antibody | Rabbit anti-BIN1, 2-5 μg/ml (datasheet A01551-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01551-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01551-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | BIN1-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: High cytoplasmic expression in CNS and skeletal muscle. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A01551-1); consider the published Tris-EDTA pH 9.0 method when adapting the HCC protocol (PMC10495885).
Section 2
What Is the Expected BIN1 Staining Pattern?
BIN1 staining should be predominantly cytoplasmic in CNS cells and skeletal myocytes (HPA tissue IHC: high cytoplasmic expression in CNS and skeletal muscle). Cytoplasmic or membrane-associated signal is consistent with BIN1 localisation; UniProt also lists the nucleus, endosomes, sarcolemma and T-tubules, and reports no transmembrane segment (UniProt O00499). Interpret tissue staining with HPA’s Enhanced IHC reliability in mind: RNA agreement is medium, and presumed off-target staining was disregarded (HPA tissue IHC).
| Strong cytoplasmic staining in skeletal myocytes or CNS glial cells; cerebellar granular cells show cytoplasmic or membrane staining (HPA tissue IHC: High). | This matches documented positive cells and compartments (HPA tissue IHC). Compare intensity within the identified cell population; a dark area containing several cell types does not establish which cells express BIN1 (general IHC practice). |
| Predominantly nuclear-only staining, with little cytoplasmic signal, in a known-positive section (HPA tissue IHC: high cytoplasmic profile). | Treat this as a compartment mismatch requiring review, not automatic proof of artefact: UniProt lists nuclear BIN1 (UniProt O00499). Check whether cytoplasmic signal is present in the expected cells and compare with a validated reference section (general IHC practice). |
| Prominent staining in esophageal, oral mucosal or vaginal squamous epithelial cells (HPA tissue IHC: Not detected in these cells). | This conflicts with the supplied IHC reference pattern and warrants a specificity check; cross-reactivity or endogenous detection activity are possible explanations, not established causes (HPA tissue IHC; general IHC practice). Reassess the cell identity before scoring (general IHC practice). |
| Color spread across cells, stroma or the whole section without a discernible cell pattern (general IHC practice). | Diffuse background cannot be scored as BIN1 localisation (general IHC practice). Check a no-primary control, blocking, washes and detection conditions; HPA reports presumed off-target staining was disregarded in its tissue assessment (HPA tissue IHC). |
| No signal in skeletal myocytes or documented positive CNS cells (HPA tissue IHC: High). | A negative result there is discordant with the HPA reference, but does not by itself identify a failed step (HPA tissue IHC; general IHC practice). Confirm the cells sampled, then review the antibody’s IHC validation and detection controls (general IHC practice). |
💡Expected BIN1 appearanceCall positive when identifiable skeletal myocytes or documented CNS cells show clear, predominantly cytoplasmic staining, with cytoplasmic or membrane staining in cerebellar granular cells (HPA tissue IHC: High); prominent signal in reference-negative squamous epithelial cells is a specificity warning (HPA tissue IHC: Not detected).
| Tissue and cell selection (HPA tissue IHC). | HPA reports High staining in skeletal myocytes, CNS glial cells and cerebellar granular cells, but Low staining in cardiomyocytes (HPA tissue IHC). Select and score the named cells; a weak heart-muscle result is less informative than a weak skeletal-muscle result (HPA tissue IHC; general IHC practice). |
| Isoform distribution (UniProt O00499). | UniProt lists 11 BIN1 isoforms and reports brain-restricted isoform IIA, while isoform BIN1 is widely expressed with highest expression in skeletal muscle (UniProt O00499). An antibody’s isoform coverage cannot be inferred from tissue staining alone; check its stated epitope before interpreting differences (general IHC practice). |
| Location and membrane association (UniProt O00499; HPA tissue IHC). | UniProt lists cytoplasm, endosome, sarcolemma and T-tubules without a transmembrane segment; HPA describes a high cytoplasmic tissue profile (UniProt O00499; HPA tissue IHC). Score visible cell-associated staining by compartment rather than assuming every membrane-shaped feature is specific (general IHC practice). |
| IHC validation and its limit (HPA antibodies; HPA tissue IHC). | HPA marks HPA003894, HPA005437 and CAB001945 Enhanced for IHC, meaning the pattern is reproduced by independent antibodies or orthogonal data (HPA antibodies). HPA still reports medium RNA consistency and disregarded presumed off-target binding; examine unexpected staining locally (HPA tissue IHC). |
| IF/ICC: what localisation is supported? (HPA subcellular). | Cytosol is the supported main location in HPA ICC-IF (HPA subcellular). That observation helps interpret an IF image, but it does not turn the tissue IHC pattern into an IF/ICC protocol or establish that every cell line stains identically (HPA subcellular; general IHC/IF practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Skeletal muscle lacks detectable myocyte staining (HPA tissue IHC: High). | The expected positive cells may be absent from the viewed area, or the staining workflow may have failed; the image alone cannot distinguish these possibilities (HPA tissue IHC; general IHC practice). | Confirm myocytes on the counterstained section, inspect a positive control, and check the catalog antibody’s IHC-P instructions and detection steps (general IHC practice). |
| CNS staining appears confined to an unexpected cell population (HPA tissue IHC: high cytoplasmic CNS profile). | Cell identification may be uncertain, or the signal may differ from HPA’s reported glial and cerebellar granular-cell patterns (HPA tissue IHC; general IHC practice). | Identify cells using morphology and anatomical context; compare the same cell type and compartment with HPA’s tissue reference before assigning positivity (HPA tissue IHC; general IHC practice). |
| Squamous epithelium stains strongly in an esophagus, oral mucosa or vagina section (HPA tissue IHC: Not detected). | Unexpected antibody binding or endogenous detection activity is possible; HPA’s presumed off-target staining caveat supports caution, not a diagnosis of the cause (HPA tissue IHC; general IHC practice). | Run a no-primary control, review blocking and detection, and compare a documented positive cell population before scoring epithelial signal as BIN1 (general IHC practice; HPA tissue IHC). |
| The entire section has diffuse chromogen signal (general IHC practice). | Background from blocking, washing or detection can obscure cellular localisation; diffuse color alone cannot establish BIN1 expression (general IHC practice). | Compare with a no-primary control, inspect wash and detection conditions, then reassess whether signal resolves into the expected cells and compartments (general IHC practice; HPA tissue IHC). |
| Staining is chiefly nuclear with little cytoplasmic signal (HPA tissue IHC: high cytoplasmic profile). | The pattern differs from HPA’s dominant tissue pattern, although nuclear localisation is listed by UniProt; its appearance alone cannot prove an artefact (HPA tissue IHC; UniProt O00499). | Review cell identity, the cytoplasmic positive control and antibody validation; record the nuclear signal separately until its specificity is supported (general IHC practice; HPA antibodies). |
| Heart muscle stains more weakly than skeletal muscle (HPA tissue IHC: Low in cardiomyocytes; High in skeletal myocytes). | This contrast is consistent with HPA’s cell-specific levels and does not alone indicate a technical failure (HPA tissue IHC). | Use the expected level for each named cell population when judging the result; investigate workflow failure if the documented High skeletal-myocyte control is also absent (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing BIN1 staining
A01551-1 · IHCIHC analysis of BIN1 using anti-BIN1 antibody (A01551-1). BIN1 was detected in a paraffin-embedded section of human appendiceal adenocarcino
A01551-1 · IHCIHC analysis of BIN1 using anti-BIN1 antibody (A01551-1). BIN1 was detected in a paraffin-embedded section of human gastric adenocarcinoma t
A01551-1 · IFIF analysis of BIN1 using anti-BIN1 antibody (A01551-1). BIN1 was detected in an immunocytochemical section of Caco-2 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136717-BIN1/tissue/cerebral+cortex#imid_2234560
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136717-BIN1/tissue/colon#imid_2234470
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136717-BIN1/tissue/liver#imid_2234487
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136717-BIN1/tissue/kidney#imid_2234488
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003894; green = BIN1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003894; green = BIN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BIN1 IHC & IF
🧪Run cerebral cortex first: glial cells should stain (HPA: High in cerebral cortex glial cells). Use esophageal squamous epithelium as the negative tissue (HPA: Not detected); on the cortex slide, cells without BIN1 staining should remain at background, but their identity as negative cells requires validation.
Positive control tissue: Caudate (Glial cells, HPA High)
Negative control tissue: Esophagus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BIN1 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control appropriate to the antibody’s clonality; and BIN1-knockout material or an immunogen-peptide competition control if available (caption: rabbit anti-BIN1 antibody). For the caption’s biotin-based DAB detection, quench endogenous peroxidase and check for endogenous biotin signal in cortex (caption: SABC with DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A01551-1 tissue-IHC caption does not state a fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 is a starting condition from the paraffin-section caption; whether BIN1 staining depends on that retrieval condition in cerebral cortex is unreported (caption: EDTA retrieval in appendiceal adenocarcinoma). The supplied evidence does not establish whether frozen sections or IF are easier; for cortex IF, check for tissue autofluorescence with a no-primary control (HPA: High in cerebral cortex glial cells; standard IF practice).
HPA tissue IHC evidence for BIN1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells type I | High | Protein (IHC) | HPA → |
Section 3
Advanced BIN1 IHC Tips
Troubleshoot BIN1 staining in paraffin sections by checking retrieval, compartment, cell identity, and antibody controls before comparing staining intensity.
Which retrieval conditions should I try first for BIN1 IHC?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet A01551-1). The selected image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A01551-1). If staining is weak, adjust heating time in small increments while keeping the buffer fixed, and compare adjacent sections in the same run (standard IHC practice). Include a skeletal-muscle positive control because myocytes show high BIN1 staining, and judge retrieval by signal in the expected cells rather than by the darkest area anywhere on the slide (HPA: high in skeletal-muscle myocytes; standard IHC practice).
Could fixation explain weak or uneven BIN1 staining?
Target-specific fixation sensitivity is unknown: the selected BIN1 caption identifies a paraffin section but does not state its fixative (datasheet A01551-1). Record the actual fixative and fixation duration for each specimen, then compare similarly processed sections before changing antibody concentration (standard IHC practice). Underfixation can compromise tissue preservation, whereas prolonged fixation can reduce epitope accessibility; neither effect has been established specifically for BIN1 here (standard IHC practice; datasheet A01551-1: fixative unreported). If staining varies across blocks, run them together with the same EDTA pH 8.0 retrieval and a skeletal-muscle control to separate processing variation from the staining run (datasheet A01551-1; HPA: high in skeletal-muscle myocytes; standard IHC practice).
Where should BIN1 staining appear within tissue cells?
Evaluate BIN1 chiefly in cytoplasmic and membrane-associated patterns, while allowing nuclear staining when the antibody and tissue controls support it (UniProt O00499: nucleus, cytoplasm, endosome, sarcolemma and T-tubule). High cytoplasmic staining in CNS and skeletal muscle provides useful tissue context (HPA: tissue IHC profile). In skeletal muscle, review signal in myocytes with attention to membrane-associated structure; BIN1 helps form T-tubules there (HPA: high in myocytes; UniProt O00499: function). A purely extracellular deposit or a uniform stain across every compartment warrants review of morphology, counterstain, and the no-primary control before it is scored as BIN1 (standard IHC practice).
Can one BIN1 antibody report every isoform equally?
Do not assume equivalent recognition across all 11 annotated BIN1 isoforms without a mapped epitope and isoform-level validation (UniProt O00499: isoforms; standard antibody-validation practice). BIN1 contains a BAR domain at residues 29–276 and an SH3 domain at 520–593; alternative splicing can change which sequence an antibody encounters (UniProt O00499: domains and isoforms). Isoform IIA is reported in brain gray matter and absent from white matter, while isoform BIN1 is widely expressed with highest expression in skeletal muscle (UniProt O00499: tissue specificity). If those regions stain differently, compare the antibody's stated immunogen with the isoforms under study before attributing the difference solely to abundance (standard antibody-validation practice).
How should I check a BIN1 pattern by multiplex IF?
Pair BIN1 with a marker identifying the expected cell type, such as a validated myocyte or glial marker selected for the specimen, and inspect both single-channel images before interpreting overlap (HPA: high in skeletal-muscle myocytes and CNS glial cells; standard IF practice). Choose fluorophores and filter sets after measuring unstained tissue autofluorescence, keeping the BIN1 channel away from its strongest background where feasible (standard IF practice). BIN1 has no transmembrane segment, so permeabilisation should be tested for access to cytosolic or intracellular-membrane epitopes rather than assumed from membrane proximity (UniProt O00499: topology and localisation; standard IF practice). An IF pattern supports the paraffin-section IHC interpretation only after its own antibody and channel controls pass (standard IF practice).
What should I change when BIN1 DAB staining is widespread?
First compare the tissue with a no-primary control and inspect whether color follows cells or pools along folds, section edges, and damaged areas (standard IHC practice). The selected BIN1 image used 10% goat-serum block, biotinylated secondary antibody, a streptavidin–biotin detection complex, and DAB (datasheet A01551-1). Check peroxidase quenching and endogenous-biotin controls when using that detection scheme, then adjust blocking, washing, or primary concentration one variable at a time (standard IHC practice; datasheet A01551-1: detection scheme). Widespread brown color should not be accepted solely because BIN1 is ubiquitous; the reported high cytoplasmic patterns and cell identities provide stronger checks (UniProt O00499: tissue specificity; HPA: tissue IHC profile).
How should I quantify BIN1 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and staining intensity on a consistent 0–3 scale (standard IHC scoring practice). An H-score sums each intensity category multiplied by its percentage of cells, giving a 0–300 range; use the same threshold and observer rules for every section (standard IHC scoring practice). For spatial questions, count BIN1-positive cells per mm² of viable, annotated tissue and normalise to the relevant cell population or tissue area (standard image-analysis practice). Analyse CNS glia and skeletal-muscle myocytes separately because both can show high signal, and record the compartment used for each score (HPA: positive cell profiles; standard IHC scoring practice).
How can I distinguish true BIN1 signal from artefact?
Give greatest weight to staining that follows expected cells and compartments: BIN1 is reported in cytoplasm and membrane-related locations, with high IHC signal in CNS glia and skeletal-muscle myocytes (UniProt O00499: localisation; HPA: positive cell profiles). Treat isolated staining in the wrong compartment or unexpected cell population as a finding to verify, especially because the HPA tissue assessment notes presumed off-target binding (HPA: reliability description; standard IHC practice). Exclude section-edge concentration and necrotic areas during review, and compare a no-primary control for detection-related color (standard IHC practice). If a disputed signal persists, repeat adjacent sections with the documented EDTA pH 8.0 retrieval and appropriate tissue controls before assigning biological meaning (datasheet A01551-1; standard IHC practice).
Boster reagents
Best BIN1 / Myc box-dependent-interacting protein 1 IHC Antibodies
The catalog lists anti-BIN1 antibodies for human, mouse and rat samples (catalog: reactivity); A01551-1 has IHC images of human paraffin sections and an IF image of Caco-2 cells (catalog: image captions).
Real IHC data
Anti-BIN1 Antibody ®
Cat # A01551-1
A01551-1 has IHC images of human appendiceal adenocarcinoma, gastric adenocarcinoma, lung cancer and brain paraffin sections, plus an IF image of Caco-2 cells (catalog: A01551-1 image captions). M01551-3 lists IHC and IF/ICC for human and mouse, while M01551-4 lists those applications for human; neither has an IHC or IF image in the payload (catalog: M01551-3 and M01551-4 applications, reactivity and image captions).
Which to pick: For tissue IHC, choose A01551-1 when an illustrated paraffin-section workflow matters: its human IHC captions report EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, but do not report the fixative (catalog: A01551-1 IHC image captions). For IF/ICC, A01551-1 has a Caco-2 image at 5 μg/ml; rabbit monoclonals M01551-3 and M01551-4 also list IF/ICC at 1:50–1:200, without images in the payload (catalog: IF image caption, clones, applications and dilutions). For work across species, A01551-1 lists human, mouse and rat reactivity; M01551-3 lists human and mouse, and M01551-4 lists human only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00499 (BIN1_HUMAN, Myc box-dependent-interacting protein 1).
- Human Protein Atlas. BIN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. BIN1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. BIN1 antibody validation summary (3 antibodies).
- High Indoleamine 2,3-Dioxygenase Expression along with Low Bridging Integrator-1 Expression in Hepatocellular Carcinoma Patients. Asian Pacific journal of cancer prevention : APJCP 2023 — PMC10495885.
- Parvalbumin Neuron-Targeted Loss of Alzheimer's Disease Risk Gene BIN1 Is Insufficient to Drive Cognitive or Network Excitability Changes. eNeuro 2026 — PMC13064429.
- Predominant expression of Alzheimer's disease-associated BIN1 in mature oligodendrocytes and localization to white matter tracts. Molecular neurodegeneration 2016 — PMC4973113.
- Increased expression of BIN1 mediates Alzheimer genetic risk by modulating tau pathology. Molecular psychiatry 2013 — PMC3807661.
- PubMed PMID:9195986 — UniProt-cited evidence.
- PubMed PMID:9182667 — UniProt-cited evidence.
- PubMed PMID:8782822 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BIN1 (Myc box-dependent-interacting protein 1). Expected localisation Predominantly cytoplasmic in CNS and skeletal muscle (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01551-1). Validated IHC antibody: A01551-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Esophagus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A01551-1); 2. Fixation: Image fixative and duration unreported (datasheet A01551-1); 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 A01551-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01551-1); 8. Primary antibody: Rabbit anti-BIN1, 2-5 μg/ml (datasheet A01551-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01551-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01551-1); Caveat: Presumed off-target binding may complicate scoring (HPA tissue IHC); Isoform / epitope: 11 isoforms; epitope coverage needs checking (UniProt); Regulation: Highest expression in brain and muscle (UniProt). Evidence: UniProt O00499, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet A01551-1, PMC10495885. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BIN1/immunohistochemistry-ihc-guide.