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BIN1 / Myc box-dependent-interacting protein 1 · IHC design guide
Design Immunohistochemistry for BIN1
Plan paraffin BIN1 IHC around the high cytoplasmic staining reported in CNS and skeletal muscle (HPA tissue IHC). Use the staining pattern and controls to assess specificity, mindful of reported off-target binding (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 | Cytoplasm in CNS and skeletal muscle (HPA tissue IHC) | |
| Staining pattern | High cytoplasmic staining in CNS glia and skeletal myocytes (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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target staining has been observed (HPA tissue IHC) | |
| Regulation | Highest expression in brain and muscle (UniProt) | |
| Isoform / epitope | 11 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended BIN1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A01551-1). The published FFPE hepatocellular carcinoma method provides one comparison protocol (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 EDTA pH 8.0 heat retrieval (datasheet A01551-1); compare Tris-EDTA pH 9.0 microwave retrieval if staining needs optimization (PMC10495885).
Section 2
What Is the Expected BIN1 Staining Pattern?
BIN1 staining should be predominantly cytoplasmic in CNS cells and skeletal myocytes (HPA: high cytoplasmic expression in CNS and skeletal muscle). BIN1 can also localise to the nucleus, endosomes, sarcolemma and T-tubules, and has no transmembrane segment (UniProt O00499: subcellular location and topology). Interpret tissue staining with caution: HPA rates its IHC profile Enhanced while reporting medium agreement with RNA data and presumed off-target staining that was disregarded (HPA: tissue IHC reliability).
| Strong cytoplasmic staining in skeletal myocytes or CNS glial cells (HPA: High in skeletal muscle myocytes and glial cells of caudate, cortex and hippocampus). | This fits the reported tissue pattern (HPA: high cytoplasmic expression in CNS and skeletal muscle). Judge positivity within the named cell population, since adjacent cells need not share its staining level (HPA: cell-specific tissue IHC results). |
| Predominantly nuclear staining, with little cytoplasmic signal in a known-positive cell population. | Review localisation and controls before scoring it as the expected pattern: HPA describes high cytoplasmic tissue expression, although UniProt also lists the nucleus as a possible location (HPA: tissue IHC profile; UniProt O00499: subcellular location). Nuclear staining alone does not establish an artefact. |
| Strong staining of esophageal, oral mucosal or vaginal squamous epithelium (HPA: Not detected in these squamous epithelial cells). | Treat this as discordant with the HPA tissue pattern and investigate antibody cross-reactivity or endogenous detection activity (HPA: tissue IHC results; standard chromogenic IHC practice). HPA reports presumed off-target binding in its reliability assessment (HPA: tissue IHC reliability). |
| Widespread, poorly localised colour across tissue structures and empty spaces. | A diffuse deposit is difficult to score as cell-specific BIN1 staining; assess background with the appropriate detection controls and inspect blocking and wash steps (standard chromogenic IHC practice). It does not reproduce the reported cell-specific cytoplasmic pattern (HPA: tissue IHC profile). |
| No staining in skeletal myocytes on a section intended as a positive reference (HPA: High in skeletal muscle myocytes). | First check tissue identity and whether the positive reference stained in the same run; then review retrieval, primary antibody, detection and counterstain conditions (standard chromogenic IHC practice). An absent signal in one run does not establish that BIN1 is absent from that tissue (HPA: tissue IHC result). |
💡Expected BIN1 appearanceCall a result positive when skeletal myocytes or the listed CNS cell populations show clear, predominantly cytoplasmic staining at the reported High level; strong staining of HPA-negative squamous epithelium or widespread unstructured colour is discordant and needs control review (HPA: tissue IHC profile and cell-specific results; standard chromogenic IHC practice).
| Cell population selected for interpretation | HPA reports High staining in skeletal myocytes, selected CNS cells and lung alveolar type I cells, but Not detected in specified squamous epithelial cells (HPA: tissue IHC results). Compare the same cell type when judging a positive or negative result. |
| Antibody validation and specificity | Three listed antibodies have Enhanced IHC validation, but HPA describes medium agreement with RNA data and disregarded presumed off-target binding (HPA: antibody validation; HPA: tissue IHC reliability). Concordant localisation and control results still matter when scoring an unfamiliar pattern. |
| Isoform and compartment context | UniProt lists 11 BIN1 isoforms and reports brain-restricted isoform IIA, while isoform BIN1 is widely expressed with its highest expression in skeletal muscle (UniProt O00499: isoforms and tissue specificity). HPA's tissue profile alone cannot assign staining to a particular isoform (HPA: tissue IHC profile). |
| Interpretation of membrane-associated signal | UniProt places BIN1 at the sarcolemma and T-tubules as well as in cytoplasm, endosomes and nucleus, despite listing no transmembrane segment (UniProt O00499: subcellular location and topology). Membrane-adjacent staining can be plausible, but assess it alongside the observed predominantly cytoplasmic IHC pattern (HPA: tissue IHC profile). |
| Retrieval and fixation evidence | The supplied HPA and UniProt records give no BIN1-specific fixation sensitivity or retrieval condition. Optimise retrieval against a known-positive tissue and matched controls as a general IHC workflow step (standard chromogenic IHC practice); do not infer a target-specific fixation effect from the tissue profile. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive skeletal muscle has no specific colour (HPA: High in skeletal muscle myocytes). | The run may have lost sensitivity through a retrieval, primary antibody or detection problem (standard chromogenic IHC practice). | Confirm tissue identity, include a same-run positive reference, and check the validated antibody's IHC conditions plus detection reagents before interpreting the sample as negative (standard chromogenic IHC practice). |
| Squamous epithelium stains strongly where HPA reports no detection (HPA: esophagus, oral mucosa and vagina). | Possible primary antibody cross-reactivity or endogenous detection activity; HPA also notes disregarded presumed off-target binding (HPA: tissue IHC reliability; standard chromogenic IHC practice). | Review a no-primary control and the relevant endogenous-activity block; compare localisation with an independently validated IHC antibody if available (standard chromogenic IHC practice; HPA: three Enhanced IHC antibodies). |
| Colour is diffuse or extends beyond recognisable cells. | Background from nonspecific reagent binding, inadequate washing or detection activity can obscure cell boundaries (standard chromogenic IHC practice). | Inspect no-primary and detection controls, washing and blocking, then score only clearly localised cellular staining (standard chromogenic IHC practice). |
| A section appears nuclear-only, unlike the expected cytoplasmic profile (HPA: high cytoplasmic expression). | UniProt permits nuclear localisation, so compartment alone cannot prove an artefact; the discrepancy may also reflect background or cell identification (UniProt O00499: subcellular location; standard IHC interpretation). | Identify the stained cell population, review counterstain and controls, and report the compartment explicitly rather than calling it the typical HPA tissue pattern (HPA: tissue IHC profile; standard IHC interpretation). |
| The sample stains weakly compared with a skeletal muscle reference (HPA: High in skeletal muscle myocytes). | Expression differs by cell type: HPA reports Low staining in cardiomyocytes, small-intestinal glandular cells, urothelial cells and breast adipocytes (HPA: tissue IHC results). | Compare like cell populations and confirm the reference works in the same run before changing the assay (HPA: cell-specific tissue IHC results; standard chromogenic IHC practice). |
| An IF/ICC image shows cytosolic BIN1 and seems different from a paraffin-section result. | HPA supports cytosolic localisation in ICC-IF, while its tissue IHC profile reports high cytoplasmic expression in CNS and skeletal muscle (HPA: subcellular ICC-IF summary; HPA: tissue IHC profile). | Interpret each result in its own preparation and named cell population; use the separate IF/ICC guide for its workflow (HPA: subcellular ICC-IF summary and tissue IHC results). |
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 caudate first and assess staining in glial cells (HPA: High in caudate glial cells). Use esophageal squamous epithelium as the negative tissue (HPA: Not detected); no cell population within caudate is established here as an internal negative, so cells lacking specific staining should show only background and should not be assumed BIN1-negative (UniProt O00499: ubiquitous expression).
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 and a rabbit isotype control matched to the catalog antibody’s clonality (hero caption: rabbit anti-BIN1; standard IHC practice). Use BIN1 knockout material or a validated peptide block as a biological specificity control (standard IHC practice). Quench endogenous peroxidase for chromogenic detection, and check caudate autofluorescence if assessing IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01551-1 paraffin-section caption does not report a fixative (hero caption). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required for every specimen (hero caption). The supplied evidence does not establish that frozen sections or IF are easier; caudate autofluorescence can complicate IF interpretation (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 type and controls before comparing chromogenic signal across samples.
Which retrieval condition should I start with for BIN1 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A01551-1). The catalog antibody produced BIN1 staining in a paraffin-embedded human appendiceal adenocarcinoma section under that condition, with primary antibody at 2 μg/ml overnight at 4°C (datasheet A01551-1). If staining is weak, first check heating consistency and section adhesion, then compare retrieval duration on serial sections while keeping detection settings fixed (standard IHC practice). Judge improvement by signal in an expected positive compartment and by background together; stronger diffuse staining alone does not establish better retrieval (HPA: high cytoplasmic expression in CNS and skeletal muscle; standard IHC practice).
How should I investigate weak BIN1 staining when fixation history is uncertain?
Target-specific BIN1 sensitivity to fixation is unknown from the supplied evidence; the paraffin-section caption does not state its fixative (datasheet A01551-1). Record each specimen’s fixative, fixation duration and processing history before comparing staining between blocks (standard IHC practice). Run sections from the blocks together using EDTA retrieval at pH 8.0 and the same antibody and detection conditions, then inspect tissue preservation alongside signal (datasheet A01551-1; standard IHC practice). If only heavily processed blocks stain poorly, report an association with processing rather than a proven BIN1-specific fixation effect, and use a known positive section to check assay performance (standard IHC practice).
Which BIN1 staining patterns should I expect, and when is nuclear signal plausible?
Expect prominent cytoplasmic staining in CNS and skeletal muscle sections, while allowing membrane-associated patterns where tissue structure supports them (HPA: high cytoplasmic expression in CNS and skeletal muscle; UniProt O00499: sarcolemma and T-tubule localisation). BIN1 is also annotated in the nucleus and endosome, whereas supported ICC/IF localisation is cytosolic, so nuclear-only chromogenic staining warrants independent confirmation (UniProt O00499: subcellular locations; HPA: cytosol supported). Compare signal with tissue architecture and a counterstain, especially in tightly packed cells (standard IHC practice). For a questionable compartment, repeat staining with a second epitope or an orthogonal assay and review the control section before assigning biological meaning (standard IHC practice).
Could isoform choice or epitope accessibility explain discordant BIN1 tissue staining?
BIN1 has 11 annotated isoforms, including brain-restricted isoform IIA and a widely expressed BIN1 isoform with highest expression in skeletal muscle (UniProt O00499: isoforms and tissue specificity). Its BAR domain spans residues 29–276, its SH3 domain spans 520–593, and several annotated phosphorylation sites lie near residues 296–307 (UniProt O00499: domains and modified residues). The supplied antibody caption does not identify the recognized epitope, so do not assign isoform specificity or phosphorylation sensitivity from a staining difference (datasheet A01551-1: epitope unreported; standard IHC practice). Check the antibody’s documented epitope and compare an independent reagent or transcript evidence before interpreting regional staining as an isoform shift (standard IHC practice).
How can I check a BIN1 IHC pattern with multiplex immunofluorescence?
Use IF/ICC as a separate confirmation assay and pair BIN1 with a validated marker for the expected cell type, such as an appropriate glial or myocyte marker in the corresponding tissue (HPA: high staining in CNS glial cells and skeletal-muscle myocytes; standard IF practice). Choose a fluorophore in a channel with low measured tissue autofluorescence, and acquire unstained and single-label controls before interpreting overlap (standard IF practice). BIN1 lacks a transmembrane segment and has supported cytosolic ICC/IF localisation, so use permeabilisation when the antibody must access an intracellular epitope (UniProt O00499: topology; HPA: cytosol supported; standard IF practice). Optimize fixation and permeabilisation in IF independently; the paraffin IHC caption supplies no IF fixation condition (datasheet A01551-1; standard IF practice).
What should I check when BIN1 DAB staining is diffuse or unusually dark?
First examine a no-primary control and verify that endogenous peroxidase was blocked before DAB development (standard chromogenic IHC practice). The documented workflow used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a biotinylated secondary with avidin-biotin detection (datasheet A01551-1). If background persists, titrate primary concentration and detection time on adjacent sections while retaining the EDTA pH 8.0 retrieval condition (datasheet A01551-1; standard IHC practice). Treat widespread staining cautiously because the HPA tissue profile reports presumed off-target binding that was disregarded in its assessment (HPA: Enhanced reliability description).
How should I score BIN1 IHC across regions with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and staining intensity as an H-score when individual cells can be resolved (standard IHC practice). For spatial patterns, count positive cells per mm² within a prespecified tissue region and normalize to the area of viable tissue or to the number of eligible cells (standard IHC practice). Keep section thickness, retrieval, imaging exposure and DAB development consistent across comparison groups (standard IHC practice). Report CNS glial cells or skeletal-muscle myocytes separately from surrounding cells because both are documented high-staining populations, and review scores against a negative control (HPA: high in CNS glial cells and skeletal-muscle myocytes; standard IHC practice).
How can I distinguish genuine BIN1 signal from chromogenic artefact?
A credible pattern should agree with the sampled cell type and an annotated compartment: BIN1 is prominent in CNS and skeletal muscle cytoplasm and can localize to sarcolemma or T-tubules (HPA: high cytoplasmic expression in CNS and skeletal muscle; UniProt O00499: subcellular locations). Check nuclear-only or anatomically unexpected staining against an independent reagent or assay, since the tissue profile notes presumed off-target binding (UniProt O00499: nuclear localisation; HPA: Enhanced reliability description; standard IHC practice). Exclude section edges and necrotic areas from scoring, and inspect a no-primary control for endogenous enzyme signal (standard chromogenic IHC practice). Interpret residual DAB staining together with morphology and the documented control pattern rather than intensity alone (standard IHC practice).
Boster reagents
Best BIN1 / Myc box-dependent-interacting protein 1 IHC Antibodies
A01551-1 has IHC images from human paraffin sections and an IF image from Caco-2 cells (catalog image captions); its listed reactivity includes human, mouse and rat (catalog reactivity).
Real IHC data
Anti-BIN1 Antibody ®
Cat # A01551-1
A01551-1 is the sole rendered card, with IHC images from human appendiceal adenocarcinoma, gastric adenocarcinoma, lung cancer and brain paraffin sections (catalog IHC captions). Its IF image uses Caco-2 cells, and its listed applications include IHC, IF and ICC (catalog IF caption; catalog applications).
Which to pick: For tissue IHC, choose A01551-1: its own caption documents human paraffin sections, EDTA retrieval at pH 8.0, and 2 μg/ml primary antibody; the fixative is unreported (A01551-1 IHC caption). For IF/ICC, A01551-1 has an IF image in Caco-2 cells using 5 μg/ml primary antibody (A01551-1 IF caption). For cross-species planning, A01551-1 lists human, mouse and rat reactivity, while rabbit monoclonals M01551-3 and M01551-4 list human/mouse and human reactivity, respectively; the supplied IHC and IF images for A01551-1 document human samples only (catalog reactivity; catalog image captions).
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 Cytoplasm 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: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target staining has been observed (HPA tissue IHC); Isoform / epitope: 11 isoforms; epitope coverage is unspecified (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.