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- Table of Contents
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).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| 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 |
| 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) |
The catalog antibody protocol is accompanied by three published RANBP1 IHC protocols for paraffin sections (PMC13480108; PMC12586255; PMC3953127).
| Sample | Paraffin-embedded mouse intestine tissues; fixative not specified (datasheet A04000-2) |
| Fixation | Image fixative and duration unreported (datasheet A04000-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A04000-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04000-2) |
| Primary antibody | Rabbit anti-RANBP1, 0.5-1μg/ml (datasheet A04000-2) |
| Primary incubation | Overnight at 4 °C (datasheet A04000-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04000-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RANBP1-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. |
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).
| 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. |
| Cell population and tissue choice | HPA 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 evidence | HPA065931 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 limits | UniProt 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). |
| Situation | Likely cause | Next 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. |
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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| 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 → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| 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 → |
Troubleshoot RANBP1 staining in paraffin sections by checking retrieval, compartment, cell type and controls before interpreting chromogenic signal.
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).
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).