RANBP1 / Ran-specific GTPase-activating protein · IHC design guide

Design Immunohistochemistry for RANBP1

Plan RANBP1 paraffin-section IHC around the cytoplasmic tissue pattern reported by HPA (HPA tissue IHC). Use the documented positive and unstained cell populations to select controls and score chromogenic staining by cell type (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for RANBP1 (IHC for RANBP1): expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A04000-2, validated IHC image, and IHC protocol steps
Printable RANBP1 IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC), antibody A04000-2, controls and protocol steps. Open the full RANBP1 IHC guide →

RANBP1 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 Cytoplasmic in testis, gut, lymphoid and hematopoietic tissues, and squamous epithelia (HPA tissue IHC)
Antigen retrieval Citrate pH 6 HIER, heat-mediated (datasheet A04000-2)
Positive control ⓘ Appendix+4 more · see all
Negative control ⓘ Adipose tissue+4 more · see all
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific)
Caveat Staining and RNA show medium consistency (HPA tissue IHC)
Regulation Expression regulation unreported (UniProt)
Isoform / epitope 2 isoforms; epitope differences are unreported (UniProt)
Section 1

Recommended RANBP1 IHC & IF Protocols

The catalog antibody protocol is accompanied by three published RANBP1 IHC protocols for paraffin sections (PMC13480108; PMC12586255; PMC3953127).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded mouse intestine tissues; fixative not specified (datasheet A04000-2)
FixationImage fixative and duration unreported (datasheet A04000-2); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: Citrate pH 6, 20 min (datasheet A04000-2)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A04000-2)
Primary antibodyRabbit anti-RANBP1, 0.5-1μg/ml (datasheet A04000-2)
Primary incubationOvernight at 4 °C (datasheet A04000-2)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A04000-2)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultRANBP1-positive staining in enterocytes of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in testis, intestinal tract, lymphoid and hematopoietic tissue, and squamous epithelia. No signal in the no-primary control.
💡Decision noteStart with heat retrieval in citrate pH 6 for the catalog antibody (datasheet: A04000-2). The ovarian study used pH 8 citrate-EDTA with a different antibody (PMC3953127).
Section 2

What Is the Expected RANBP1 Staining Pattern?

RANBP1 staining is expected mainly in the cytoplasm: HPA reports cytoplasmic tissue staining and an enhanced cytosolic ICC-IF location (HPA: tissue IHC; HPA: subcellular). In paraffin sections, look for staining in reported positive cells such as appendix enterocytes, intestinal endocrine cells, breast glandular cells, and ovary follicle cells (HPA: High). Tissue IHC reliability is Supported, with medium consistency against RNA data (HPA: tissue IHC). RANBP1 has no transmembrane segment (UniProt P43487 topology).

What am I looking at on my slide?
Cytoplasmic chromogen is clear in appendix enterocytes or intestinal endocrine cells, while cell boundaries and counterstained nuclei remain readable.This fits the reported location and cell pattern (HPA: cytoplasmic tissue expression; HPA: High in appendix enterocytes and intestinal endocrine cells). Score the named cell population, intensity, and proportion separately; a tissue label alone does not establish that every cell should stain (standard IHC practice).
The dominant signal is nuclear, membranous, or confined to extracellular material.Question that result: HPA reports cytoplasmic IHC staining and an enhanced cytosolic ICC-IF location (HPA: tissue IHC; HPA: subcellular). UniProt lists no transmembrane segment (UniProt P43487 topology). Review controls and detection before calling it RANBP1; this pattern alone cannot identify the artefact.
Strong signal appears in adipocytes or kidney glomerular cells, with little signal in the expected positive population.HPA reports RANBP1 as Not detected in those named cells (HPA: adipocytes; HPA: kidney glomerular cells). Check the exact cell identity and controls for cross-reactivity or endogenous detection activity (standard IHC practice). HPA reports low tissue RNA specificity, so an unexpected cell signal is a reason to verify, not proof of a false positive (HPA: tissue IHC).
Brown color spreads across the section, obscures cell boundaries, or covers tissue and empty spaces similarly.Treat this as background until a cell-specific cytoplasmic pattern is resolved (standard chromogenic IHC practice). HPA's observation concerns cytoplasmic expression in defined tissue populations, not uniform section-wide color (HPA: tissue IHC). Compare negative controls before scoring intensity or proportion.
Appendix enterocytes show no detectable cytoplasmic stain in an otherwise interpretable section.That conflicts with an HPA High observation, but HPA rates tissue IHC Supported with medium RNA agreement (HPA: appendix enterocytes; HPA: tissue IHC reliability). Verify tissue identity and assay controls, then review the antibody-specific IHC-P procedure; one negative section cannot establish absence of RANBP1.
💡Expected RANBP1 appearanceA convincing positive is cell-resolved cytoplasmic staining, potentially strong in HPA High populations such as appendix enterocytes (HPA: tissue IHC); dominant nuclear color or diffuse section-wide deposit is a suspect result requiring controls (HPA: subcellular; standard IHC practice).
How each factor affects the staining
Cell population and tissue choiceHPA calls several specific populations High, including breast glandular cells and ovary follicle cells, while adipocytes and kidney glomerular cells are Not detected (HPA: tissue IHC). Evaluate the named cells within each section; do not turn those observations into a claim that the entire tissue is uniformly positive or negative.
Strength of antibody evidenceHPA065931 has Supported IHC validation; HPA065868 has no listed IHC validation status (HPA: antibodies). HPA also describes medium agreement between tissue staining and RNA expression (HPA: tissue IHC reliability). Use this evidence to set confidence in the expected pattern, without treating either antibody as independently proven for every tissue.
Topology, isoforms, and epitope limitsUniProt lists two RANBP1 isoforms, a RanBD1 domain at residues 26–164, and no transmembrane segment (UniProt P43487). The payload does not map either antibody's epitope, so these sequence features do not predict isoform coverage, retrieval response, or a distinct membrane signal.
IF/ICC: should its location match IHC?The expected compartment agrees: HPA assigns an enhanced cytosolic ICC-IF location, consistent with its cytoplasmic tissue IHC profile (HPA: subcellular; HPA: tissue IHC). HPA lists Enhanced ICC validation for both antibodies, but that does not supply an IF protocol or establish identical staining intensity across preparations (HPA: antibodies).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
A reported High population is blank, including appendix enterocytes.The observation may reflect assay performance or section selection; HPA's High call is an observed cell pattern, not a guarantee for every section (HPA: appendix; HPA: tissue IHC reliability).Confirm the cell population, inspect tissue integrity and control slides, then check the chosen antibody's documented IHC-P retrieval, dilution, incubation, and detection settings (standard IHC practice). Do not infer RANBP1-specific fixation sensitivity from these data.
Signal is mainly nuclear or outlines membranes.The compartment conflicts with HPA's cytoplasmic IHC and enhanced cytosolic ICC-IF reports; UniProt provides no membrane-spanning topology (HPA: tissue IHC; HPA: subcellular; UniProt P43487 topology).Recheck microscopy focus, counterstain, and cell boundaries; compare a no-primary control and an expected positive population (standard IHC practice). Record the discordance rather than assigning a new RANBP1 location from one slide.
Adipocytes or kidney glomerular cells stain strongly.Those specific populations are listed as Not detected by HPA, so strong staining is discordant with the reference pattern (HPA: adipose tissue; HPA: kidney). The color alone cannot distinguish target signal from cross-reactivity or detection background.Confirm the named cells, inspect the no-primary control, and compare staining in a reported High population on the same run (standard IHC practice; HPA: tissue IHC). Investigate persistent cell-specific discordance before scoring it as RANBP1.
Uniform brown haze obscures the expected cells.Diffuse color prevents a reliable assessment of HPA's cell-specific cytoplasmic pattern (HPA: tissue IHC); several blocking, washing, or detection issues can produce background in chromogenic IHC (standard IHC practice).Review no-primary and detection controls, blocking, washes, and development time; adjust the workflow only against its controls (standard IHC practice). Reassess compartment and cell identity after the background clears.
Two antibodies give different IHC patterns.Their supplied IHC evidence differs: HPA065931 is Supported, whereas HPA065868 has no listed IHC status (HPA: antibodies). The record supplies no epitope map or target-specific explanation for a mismatch.Compare each reagent's documented IHC use and controls, then judge both against the same HPA cytoplasmic, cell-specific reference pattern (HPA: antibodies; HPA: tissue IHC). Do not resolve the discrepancy by assuming an isoform-specific stain.
ICC-IF looks cytosolic, but paraffin IHC has weak or patchy staining.The assays use different preparations and readouts (standard IHC/IF practice). HPA reports enhanced cytosolic ICC-IF localization and Supported tissue IHC reliability, which describe related but distinct evidence (HPA: subcellular; HPA: tissue IHC).Keep the IF result as a compartment check; assess the IHC section using its own positive cells, controls, and antibody-specific IHC-P procedure (standard IHC practice; HPA: tissue IHC). Do not use ICC-IF validation as an IHC protocol.

Sample controls for RANBP1 IHC & IF

🧪Run appendix first and require staining in enterocytes (HPA: High in appendix enterocytes). Use adipose tissue adipocytes as a negative comparator (HPA: Not detected in adipocytes); on the appendix slide, unstained neighboring cells can indicate local background, but no specific appendix cell type is documented as an internal negative (HPA: appendix enterocytes High).
Positive control tissue: Appendix (Enterocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RANBP1 in HEK293, Rh30, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control, and RANBP1 knockout material if available (caption: rabbit primary; standard IHC practice). For the biotin-based DAB method, block endogenous peroxidase and biotin to assess background (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A04000-2 tissue-IHC caption does not state the fixative (caption: fixative unreported). Heat retrieval in citrate buffer at pH 6 for 20 minutes produced staining in paraffin-embedded mouse intestine; retrieval dependence and whether frozen sections are easier remain unreported (caption: mouse intestine IHC). IF/ICC images support cytosolic localization, but do not establish that IF is easier; intestinal sections using biotin-based detection warrant attention to endogenous biotin background (HPA: cytosol; caption: SABC detection; standard IHC practice).

HPA tissue IHC evidence for RANBP1

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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

TissueCell typeLevelEvidenceSource
Appendix Enterocytes High Protein (IHC) HPA →
Breast Glandular cells High Protein (IHC) HPA →
Colon Endocrine cells High Protein (IHC) HPA →
Duodenum Endocrine cells High Protein (IHC) HPA →
Endometrium Glandular cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes Not detected Protein (IHC) HPA →
Caudate Glial cells Not detected Protein (IHC) HPA →
Cerebellum Cells in granular layer Not detected Protein (IHC) HPA →
Cerebral cortex Endothelial cells Not detected Protein (IHC) HPA →
Gallbladder Glandular cells Not detected Protein (IHC) HPA →
Section 3

Advanced RANBP1 IHC Tips

Troubleshoot RANBP1 staining in paraffin sections by checking retrieval, compartment, cell type and controls before interpreting chromogenic signal.

Which retrieval conditions should I start with for RANBP1 IHC?
Start with heat-mediated antigen retrieval in citrate buffer at pH 6 (datasheet A04000-2). The catalog antibody's mouse intestine paraffin-section example used this buffer for 20 minutes, followed by 10% goat serum blocking and 1 μg/mL primary antibody overnight at 4°C (datasheet A04000-2). If staining is weak, check that sections reached the intended retrieval temperature and that cooling and washes were consistent across slides (standard IHC practice). Compare any adjusted retrieval time against a section processed with the documented conditions, because stronger heating can increase background or damage tissue morphology (standard IHC practice).
Could fixation explain weak RANBP1 staining in my paraffin sections?
Target-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence; the selected mouse intestine paraffin-section caption does not state a fixative (datasheet A04000-2). Record fixative, time before fixation, fixation duration and processing conditions for each specimen so weak staining can be investigated systematically (standard IHC practice). Compare a weak section with a similarly processed positive-control section in the same staining run, keeping retrieval and antibody conditions matched (standard IHC practice). If morphology is compromised or staining differs by specimen preparation, test processing variables on matched material before attributing the difference to RANBP1 abundance (standard IHC practice).
Should I accept nuclear staining as the main RANBP1 signal?
Expect predominantly cytoplasmic staining in tissue IHC, consistent with the reported tissue profile, and use cytosolic staining as a comparison from cell imaging (HPA: tissue IHC profile; HPA: cytosol, enhanced). RANBP1 participates in RAN-dependent nucleocytoplasmic transport, but that function alone does not establish nuclear antibody staining as specific (UniProt P43487 function). Review the chromogen distribution within intact cells at higher magnification, using a counterstain to distinguish nuclei from surrounding cytoplasm (standard IHC practice). If nuclear staining predominates, compare positive-control tissue and a no-primary control under identical detection conditions before scoring it as RANBP1 (standard IHC practice).
How could RANBP1 isoforms or epitope location affect IHC staining?
RANBP1 has 2 listed isoforms and a Ran-binding domain spanning residues 26–164, but the supplied evidence does not map this antibody's epitope (UniProt P43487 isoforms and domains; datasheet A04000-2). Therefore, a staining difference cannot be assigned to one isoform without antibody epitope information or an independent isoform-specific method (standard IHC interpretation). Several modified residues are reported near the N terminus, including phosphorylation at residues 13, 18 and 21 (UniProt P43487 modified residues). If results vary across preparations, compare matched sections and retrieval conditions before proposing that a modification changes antibody binding (standard IHC practice).
How should I follow up a chromogenic RANBP1 result with multiplex IF?
Treat IF/ICC as a separate validation workflow while using the cytosolic localisation evidence to guide image review (HPA: cytosol, enhanced). Multiplex RANBP1 with a validated marker for the cell population being assessed, such as an enterocyte marker when examining the appendix enterocytes reported as high in tissue IHC (HPA: High in appendix enterocytes; standard IF practice). Choose fluorophores after checking autofluorescence in an unstained section, and place the weaker signal in a channel with lower tissue background (standard IF practice). Because RANBP1 has no transmembrane segment and is reported in the cytosol, use controlled permeabilisation to expose an intracellular epitope, then verify localisation with single-stain controls (UniProt P43487 topology; HPA: cytosol, enhanced; standard IF practice).
What should I check when RANBP1 DAB staining looks diffuse?
Inspect a no-primary control to assess signal from the secondary antibody, detection reagents or tissue itself before changing the RANBP1 primary concentration (standard IHC practice). The documented example uses biotinylated goat anti-rabbit secondary antibody, a streptavidin-biotin detection complex and DAB, so check detection-system background when using that workflow (datasheet A04000-2; standard IHC practice). Include an appropriate peroxidase block and examine whether residual enzyme activity contributes to brown signal (standard chromogenic IHC practice). If background persists, compare blocking, washing and primary-antibody conditions on adjacent sections while retaining a positive-control tissue, and judge improvement by preserved cellular staining and morphology (standard IHC practice).
How should I quantify variable RANBP1 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, because reported RANBP1 staining differs among cell types within and across tissues (HPA: tissue IHC profile). For cytoplasmic DAB staining, record the percentage of positive cells and an intensity score, then calculate an H-score if the categories are applied consistently (HPA: cytoplasmic tissue profile; standard IHC scoring practice). If counting positive cells by area, report density per mm² of viable tissue and normalise comparisons to the same cell population or sampled compartment (standard IHC quantification practice). Keep retrieval, imaging, threshold and counterstain settings consistent, and exclude folds and necrotic areas using predefined rules (standard IHC quantification practice).
How can I distinguish convincing RANBP1 staining from artefact?
Give more weight to staining in intact cell cytoplasm that follows the reported tissue distribution than to isolated nuclear or extracellular deposits (HPA: cytoplasmic tissue profile; HPA: cytosol, enhanced; standard IHC interpretation). For example, high staining in appendix enterocytes is supported, whereas staining in adipocytes is reported as not detected; interpret either observation within its specimen and assay context (HPA: High in appendix enterocytes; HPA: Not detected in adipocytes). Exclude section edges, folds and necrotic regions, where uneven reagent access or damaged tissue can mimic a biological pattern (standard IHC practice). Check no-primary and detection controls for endogenous enzyme or reagent-associated DAB signal before calling an unusual compartment or cell type positive (standard chromogenic IHC practice).
Boster reagents

Best RANBP1 / Ran-specific GTPase-activating protein IHC Antibodies

Validated RANBP1 antibodies have IHC data from paraffin sections of mouse and rat intestine and human intestinal and lung cancers (A04000-2 image captions), plus IF data from A431 and U2OS cells (catalog IF captions).

Real IHC data IHC analysis of RanBP1 using anti-RanBP1 antibody (A04000-2). RanBP1 was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-RanBP1 Antibody (A04000-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-RanBP1 Antibody ®
Cat # A04000-2
Real IF data IF analysis of RANBP1 using anti-RANBP1 antibody (A04000-3). RANBP1 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-RANBP1 Antibody (A04000-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Anti-RANBP1 Antibody ®
Cat # A04000-3

A04000-2 will render with paraffin-section IHC data from mouse and rat intestine and human intestinal and lung cancers; its IF caption also shows A431 cells (A04000-2 image captions). A04000-3 will render with IF/ICC data from U2OS cells and has no listed IHC application or IHC image (A04000-3 applications and image captions).

Which to pick: Choose A04000-2 for tissue IHC: its IHC captions document citrate retrieval in paraffin sections, while the fixative is unreported (A04000-2 IHC captions). For IF/ICC, either SKU has cell-image data; A04000-3 is listed as polyclonal and has a U2OS IF image (catalog dilution notes; A04000-3 IF caption), while A04000-2 has an A431 IF image (A04000-2 IF caption). Choose A04000-2 when human, mouse and rat reactivity matters (A04000-2 catalog reactivity).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P43487 (RANG_HUMAN, Ran-specific GTPase-activating protein).
  2. Human Protein Atlas. RANBP1 tissue IHC expression (reliability: Supported).
  3. Human Protein Atlas. RANBP1 subcellular location (ICC-IF): Localized to the cytosol..
  4. Human Protein Atlas. RANBP1 antibody validation summary (2 antibodies).
  5. Increased cathepsin D protein expression is a biomarker for osteosarcomas, pulmonary metastases and other bone malignancies. Oncotarget 2015 — PMC4599286.
  6. RANBP1 promotes oral cancer progression through activating YAP1 and NDUFB3 derived from the database of single-cell sequencing. Cancer cell international 2026 — PMC13480108.
  7. RANBP1 promotes immune evasion in triple-negative breast cancer by suppressing T cell infiltration via the miR-769-5p/PRUNE2 axis. Discover oncology 2025 — PMC12586255.
  8. RAN nucleo-cytoplasmic transport and mitotic spindle assembly partners XPO7 and TPX2 are new prognostic biomarkers in serous epithelial ovarian cancer. PloS one 2014 — PMC3953127.
  9. PubMed PMID:7882974 — UniProt-cited evidence.
  10. PubMed PMID:7616957 — UniProt-cited evidence.
  11. PubMed PMID:15461802 — UniProt-cited evidence.