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RANBP3 / Ran-binding protein 3 · IHC design guide
Design Immunohistochemistry for RANBP3
RANBP3 shows widespread nuclear and cytoplasmic tissue staining (HPA tissue IHC). This paraffin IHC guide covers the catalog antibody’s 2–5 μg/ml working range (datasheet A06252-1) and interpretation of nuclear signal in light of phosphorylation-dependent import (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RANBP3 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 | Nuclear and cytoplasmic in tissue (HPA tissue IHC) | |
| Staining pattern | Many cell types show nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06252-1) | |
| Positive control | Epididymis+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Broad staining limits negative-tissue controls (HPA tissue IHC) | |
| Regulation | Ser-126 phosphorylation promotes nuclear import (UniProt) | |
| Isoform / epitope | Three isoforms; confirm antibody epitope coverage (UniProt) |
Section 1
Recommended RANBP3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A06252-1) with the published RanBP3 tissue microarray protocol (PMC9731691 methods).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06252-1) |
| Fixation | Image fixative and duration unreported (datasheet A06252-1); 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 A06252-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06252-1) |
| Primary antibody | Rabbit anti-RANBP3, 2-5 μg/ml (datasheet A06252-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06252-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06252-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RANBP3-positive staining in glandular cells of epididymis (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A06252-1).
Section 2
What Is the Expected RANBP3 Staining Pattern?
RANBP3 is found in the nucleus and cytoplasm and has no transmembrane segment (UniProt Q9H6Z4). In tissue sections, expect widespread nuclear and cytoplasmic staining, including high staining in epididymal glandular cells and medium staining in several other listed cell types (HPA tissue IHC). HPA rates the tissue pattern Enhanced because antibody staining is highly consistent with RNA expression data (HPA tissue IHC).
| Nuclear and cytoplasmic signal appears in many cell types; epididymal glandular cells stain strongly. | This fits the ubiquitous tissue pattern and its listed high-staining example (HPA tissue IHC). Judge intensity within identified cells, since HPA also reports low staining in some cell types; a uniformly strong field is not required. |
| Signal appears mainly at the cell surface or in extracellular material, with little nuclear or cytoplasmic staining. | That distribution conflicts with the reported compartments and lack of a transmembrane segment (UniProt Q9H6Z4; HPA tissue IHC). Check morphology and staining controls before interpreting it as RANBP3; localization alone cannot identify the artefact. |
| The strongest signal is in an unexpected cell type, while the listed cells in the same section stain weakly. | For example, strong smooth muscle staining alongside weak expected glandular staining conflicts with HPA’s listed low smooth muscle result and glandular examples (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, then compare controls. |
| Color spreads across tissue structures or appears where cellular boundaries cannot be resolved. | Diffuse background prevents a reliable nuclear and cytoplasmic call. Review blocking, washes, detection controls and counterstain as general IHC practice; this appearance cannot establish a RANBP3-specific cause (general IHC practice). |
| No signal is visible in epididymal glandular cells. | That conflicts with HPA’s high-staining example (HPA tissue IHC). Confirm the cell type and examine a working positive section and detection controls before calling the sample negative; HPA supplies no universally negative tissue here. |
💡Expected RANBP3 appearanceCall a convincing positive when identifiable cells show nuclear and cytoplasmic signal, with high staining in epididymal glandular cells (HPA tissue IHC); isolated surface or extracellular color is discordant (UniProt Q9H6Z4).
| Tissue and cell type (HPA tissue IHC) | HPA describes ubiquitous staining but lists high epididymal glandular staining, several medium examples and specific low-staining cell types. Compare like cells when judging intensity; the record supplies no negative tissue. |
| Compartment and regulated import (UniProt Q9H6Z4) | RANBP3 occupies nucleus and cytoplasm; phosphorylation at Ser-126 promotes KPNA4-dependent nuclear import. Nuclear-to-cytoplasmic balance may vary, but these facts supply no IHC intensity threshold or fixation effect. |
| IHC evidence (HPA tissue IHC; HPA antibodies) | The tissue profile is rated Enhanced for consistency with RNA data. HPA043375 and HPA043389 each have Enhanced IHC validation; that supports pattern confidence without proving every signal in a new specimen is specific. |
| Isoforms and processing (UniProt Q9H6Z4) | Three isoforms are listed, with no signal peptide or propeptide and a reported chain spanning residues 2–567. These facts do not identify the catalog antibody’s epitope or establish isoform-specific staining. |
| IF/ICC reference (HPA subcellular) | HPA reports enhanced nucleoplasmic localization in ICC-IF images from A-431, U-251MG and U2OS. This is a compartment reference for the separate IF/ICC guide, not an IHC protocol or a tissue-intensity standard. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected epididymal glandular staining is absent (HPA tissue IHC). | The target-specific cause is undetermined; failed detection or misidentified cells are possibilities (general IHC practice). | Confirm glandular morphology, inspect a known-working positive section, and check primary-antibody and detection steps before scoring absence (general IHC practice). |
| Signal is confined to a membrane-like rim or extracellular space. | The location conflicts with nuclear and cytoplasmic RANBP3 and its lack of a transmembrane segment (UniProt Q9H6Z4). | Review the no-primary control and tissue morphology, then assess whether intracellular staining can be resolved (general IHC practice). |
| Unexpected cells dominate while expected cells are weak. | Cross-reactivity or endogenous detection activity is possible; HPA lists cell-specific intensity differences but cannot diagnose this slide (HPA tissue IHC). | Compare the same cell types in a positive reference and inspect detection controls before assigning the strong signal to RANBP3 (general IHC practice). |
| Diffuse chromogen obscures nuclei and cytoplasm. | Background can reflect a staining workflow problem; the supplied sources do not establish a RANBP3-specific mechanism (general IHC practice). | Review blocking, washes and detection controls, and use the counterstain to reassess cellular boundaries (general IHC practice). |
| Only faint staining appears in a low-listed cell type. | Low staining is reported for selected cells, including smooth muscle cells and liver cholangiocytes (HPA tissue IHC). | Evaluate an HPA-listed high or medium cell type before treating the faint result as assay failure; avoid a universal positive cutoff (HPA tissue IHC). |
| What should the separate IF/ICC guide expect? | HPA reports enhanced nucleoplasmic localization in ICC-IF, while UniProt also lists cytoplasm and nucleus (HPA subcellular; UniProt Q9H6Z4). | Use nucleoplasmic signal as the IF/ICC reference and interpret any cytoplasmic signal in context; consult that guide for its own workflow (HPA subcellular; UniProt Q9H6Z4). |
Experiment evidence
IHC & IF images showing RANBP3 staining
A06252-1 · IHCIHC analysis of RANBP3 using anti-RANBP3 antibody (A06252-1). RANBP3 was detected in a paraffin-embedded section of human breast cancer tiss
A06252-1 · IFIF analysis of RANBP3 using anti-RANBP3 antibody (A06252-1) and anti-Tubulin Alpha antibody (M03989-3). RANBP3 was detected in immunocytoche
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000031823-RANBP3/tissue/cerebral+cortex#imid_11962447
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000031823-RANBP3/tissue/colon#imid_11962373
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000031823-RANBP3/tissue/liver#imid_11962382
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000031823-RANBP3/tissue/kidney#imid_11962391
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043375; green = RANBP3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043375; green = RANBP3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RANBP3 IHC & IF
🧪Run epididymis first; its glandular cells should stain (HPA: High in epididymis glandular cells). HPA detects RANBP3 in all 45 scored tissues, so there is no HPA negative tissue; use no-primary and isotype controls, and expect little background in adjacent cells on the positive slide without treating those cells as confirmed biological negatives (HPA: no negative tissue rows).
Positive control tissue: Epididymis (Glandular cells, HPA High)
Negative control tissue: None in HPA: RANBP3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RANBP3 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a rabbit isotype control matched to the primary antibody’s class and clonality where known (caption: rabbit primary antibody); use RANBP3 knockout material or a validated peptide block as a biological specificity control (standard IHC practice). Quench endogenous peroxidase and inspect glandular lumens for nonspecific DAB deposit (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: The selected paraffin-section caption uses heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval is required; its fixative is unreported (caption: paraffin section and EDTA retrieval; caption: fixative not stated). No target-specific fixation window or fixation effect is reported in the supplied evidence, and there is no basis here to say frozen sections or IF are easier (supplied target/application evidence). Inspect epididymal glandular lumens for staining artefact when scoring glandular cells (HPA: High in epididymis glandular cells; standard IHC practice).
HPA tissue IHC evidence for RANBP3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RANBP3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RANBP3 IHC Tips
Troubleshoot RANBP3 staining in paraffin sections by checking retrieval, compartment pattern and cell-specific signal before comparing chromogenic scores (datasheet A06252-1; HPA tissue IHC).
Which retrieval should I start with when RANBP3 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06252-1). The published product image used that retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C, so reproduce those conditions when assessing a weak result (datasheet A06252-1). Include a positive control section in each run, and compare nuclear and cytoplasmic staining within the same cell population (standard IHC practice; HPA tissue IHC). If signal remains weak, vary heating time on matched sections while holding antibody concentration and detection constant, then reject conditions that increase diffuse background (standard IHC practice).
How should I troubleshoot variable RANBP3 staining when fixation history is uncertain?
Target-specific sensitivity to fixation is unknown because the selected tissue-IHC caption does not state a fixative (datasheet A06252-1). Record the available processing history for every block, and compare sections of similar thickness, age and processing in the same staining run (standard IHC practice). Keep heat-mediated EDTA retrieval at pH 8.0 and the primary concentration at 2 μg/ml while assessing processing-related variation (datasheet A06252-1; standard IHC practice). Use a positive control with documented processing, and interpret a uniformly weak specimen cautiously when its processing history differs from the control (standard IHC practice).
What compartment pattern should I expect for RANBP3 in chromogenic tissue IHC?
Assess both nuclear and cytoplasmic staining because tissue IHC reports ubiquitous expression in both compartments (HPA tissue IHC). Nucleoplasmic localization is supported by subcellular imaging, while the protein record also lists cytoplasmic and nuclear localization (HPA subcellular; UniProt Q9H6Z4). Nuclear import is promoted by phosphorylation at Ser-126 and depends on KPNA4, so compartment balance may vary biologically (UniProt Q9H6Z4). Score nuclear and cytoplasmic signal separately in defined cell populations, checking that staining follows cell boundaries and nuclear counterstain rather than folds or tissue edges (standard IHC practice). Do not require an exclusively nuclear pattern as the acceptance criterion (HPA tissue IHC).
Could RANBP3 isoforms or epitope accessibility explain discrepant IHC patterns?
RANBP3 has 3 recorded isoforms, and its Ran-binding domain spans residues 378–518 (UniProt Q9H6Z4). The supplied product caption does not identify the antibody epitope, so staining cannot be assigned to a particular isoform from this evidence (datasheet A06252-1; UniProt Q9H6Z4). Compare matched sections using the same EDTA pH 8.0 retrieval and 2 μg/ml primary concentration before attributing differences to epitope accessibility (datasheet A06252-1; standard IHC practice). Document any alternate antibody's stated epitope and validation separately, and require a comparable cellular pattern before interpreting differences as isoform-specific (standard IHC practice; HPA tissue IHC).
How can I compare RANBP3 immunofluorescence with this chromogenic IHC pattern?
Treat IF as a separate validation workflow: the selected antibody evidence describes paraffin-section chromogenic IHC with DAB, while subcellular imaging reports nucleoplasmic RANBP3 (datasheet A06252-1; HPA subcellular). For multiplex IF, pair RANBP3 with a marker identifying the expected cell population, such as a glandular-cell marker when examining epididymis, and confirm each antibody independently (HPA tissue IHC; standard IF practice). Choose fluorophores after checking tissue autofluorescence and single-color controls, favoring a spectral channel with cleaner contrast (standard IF practice). Optimize permeabilisation for access to the intracellular nuclear or cytoplasmic epitope, then compare compartment-resolved IF signal with matched IHC sections (UniProt Q9H6Z4; standard IF practice).
What should I change when RANBP3 DAB staining is diffuse or obscures nuclei?
First inspect a no-primary control to distinguish antibody-associated staining from detection-system or tissue background (standard IHC practice). The product image used 10% goat serum blocking, 2 μg/ml rabbit primary overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A06252-1). Apply an appropriate peroxidase block and check that DAB development preserves visible nuclear counterstain; these are general chromogenic workflow controls (standard IHC practice). If background persists, titrate primary concentration and development time on adjacent sections while retaining EDTA pH 8.0 retrieval and a positive control (datasheet A06252-1; standard IHC practice).
How should I quantify RANBP3 when nuclear and cytoplasmic signals differ? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring because RANBP3 has nuclear and cytoplasmic tissue staining (HPA tissue IHC). For each compartment, record the percentage of positive cells and staining intensity, then calculate a compartment-specific H-score using the same thresholds across slides (standard IHC practice). Normalize positive-cell counts to all evaluable cells of that population, or report positive-cell density per mm² of viable analyzed tissue when cell counts are impractical (standard IHC practice). Exclude necrosis, folds and section edges from the analysis mask, and process compared slides with the same retrieval and DAB development conditions (standard IHC practice; datasheet A06252-1).
How do I distinguish genuine RANBP3 staining from a misleading positive?
A plausible positive follows intact cells and can involve nuclei and cytoplasm; nucleoplasmic enrichment also agrees with subcellular imaging (HPA tissue IHC; HPA subcellular). Check the identified cell population against available tissue context, such as high staining in epididymal glandular cells, without treating that example as a universal intensity standard (HPA tissue IHC). Reject signal concentrated at section edges, folds or necrotic areas, and investigate deposits that do not follow cellular outlines (standard IHC practice). Compare a no-primary control and peroxidase-blocked section when DAB persists unexpectedly, because endogenous enzyme activity can mimic chromogenic signal (standard IHC practice).
Boster reagents
Best RANBP3 / Ran-binding protein 3 IHC Antibodies
A06252-1 has documented IHC in human paraffin sections and ICC/IF in U2OS cells (catalog image captions). Its listed reactivity is Human, Mouse and Rat (catalog reactivity).
Real IHC data
Anti-RANBP3 Antibody ®
Cat # A06252-1
The rendered SKU is A06252-1, with an IHC image from a human breast cancer paraffin section (A06252-1 IHC caption). Its ICC/IF image shows RANBP3 staining in U2OS cells (A06252-1 IF caption).
Which to pick: For tissue IHC, choose A06252-1: its own image documents a human breast cancer paraffin section with EDTA pH 8 retrieval and 2 μg/ml primary antibody; the fixative is unreported (A06252-1 IHC caption). For IF/ICC, A06252-1 has a U2OS image using 5 μg/ml, while M06252 lists ICC/IF but has no supplied IF image (A06252-1 IF caption; M06252 catalog applications and image alts). For cross-species selection, A06252-1 lists Human, Mouse and Rat reactivity; M06252 is a rabbit monoclonal, clone 26R52, listing Human and Mouse reactivity (catalog host, clone and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H6Z4 (RANB3_HUMAN, Ran-binding protein 3).
- Human Protein Atlas. RANBP3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RANBP3 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RANBP3 antibody validation summary (2 antibodies).
- Blockade of Nuclear β-Catenin Signaling via Direct Targeting of RanBP3 with NU2058 Induces Cell Senescence to Suppress Colorectal Tumorigenesis. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2022 — PMC9731691.
- Pegylated interferon alpha targets Wnt signaling by inducing nuclear export of β-catenin. Journal of hepatology 2011 — PMC3052972.
- PubMed PMID:9637251 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RANBP3 (Ran-binding protein 3). Expected localisation Nuclear and cytoplasmic in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06252-1). Validated IHC antibody: A06252-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Epididymis (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06252-1); 2. Fixation: Image fixative and duration unreported (datasheet A06252-1); 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 A06252-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06252-1); 8. Primary antibody: Rabbit anti-RANBP3, 2-5 μg/ml (datasheet A06252-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06252-1). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Broad staining limits negative-tissue controls (HPA tissue IHC); Isoform / epitope: Three isoforms; confirm antibody epitope coverage (UniProt); Regulation: Ser- 126 phosphorylation promotes nuclear import (UniProt). Evidence: UniProt Q9H6Z4, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A06252-1, PMC9731691. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RANBP3/immunohistochemistry-ihc-guide.