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RBM15 / RNA-binding protein 15 · IHC design guide
Design Immunohistochemistry for RBM15
Plan RBM15 paraffin-section IHC around its variable nuclear staining (HPA tissue IHC). The guide highlights high-staining Purkinje cells and colonic glandular cells for tissue selection (HPA tissue IHC), with a catalog antibody starting range of 2–5 μg/ml (datasheet A04395).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RBM15 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 | Variable nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Variable nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04395) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Soft tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04395) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Variable by tissue; marrow RNA enhanced (HPA tissue RNA) | |
| Isoform / epitope | Four isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended RBM15 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet A04395). These tables summarize published RBM15 IHC in thyroid, lung cancer, and mouse liver tissues (PMC9664221; PMC11831794; PMC9942341).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A04395) |
| Fixation | Image fixative and duration unreported (datasheet A04395); 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 A04395); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04395) |
| Primary antibody | Rabbit anti-RBM15, 2-5 μg/ml (datasheet A04395) |
| Primary incubation | Overnight at 4 °C (datasheet A04395) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04395) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RBM15-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression at variable levels in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A04395). The mouse liver study used microwave citrate retrieval (PMC9942341).
Section 2
What Is the Expected RBM15 Staining Pattern?
RBM15 should appear predominantly in nuclei: HPA reports variable nuclear staining in most tissues (HPA tissue IHC: Supported), while UniProt places RBM15 in nuclear speckles, nucleoplasm, envelope and membrane (UniProt Q96T37). Strong examples include intestinal glandular cells, kidney collecting ducts and placental decidual cells (HPA tissue IHC: High). RBM15 has no transmembrane segment (UniProt Q96T37 topology).
| Clear nuclear staining in colon or duodenal glandular cells, with variation among cells. | This fits the reported pattern: both cell populations stain at a High level, and nuclear expression varies across tissues (HPA tissue IHC: High in colon and duodenum; variable nuclear expression). Judge signal in the named cells, rather than expecting every cell in the section to match. |
| Cytoplasmic or surface staining dominates while nuclei remain pale. | That distribution conflicts with the nuclear tissue pattern and the reported nuclear locations (HPA tissue IHC: nuclear expression; UniProt Q96T37 subcellular location). Review morphology, counterstain alignment and detection background before calling it RBM15. A nuclear rim can be compatible with UniProt's envelope annotation, but diffuse cytoplasm alone is unconvincing. |
| Strong staining appears in soft tissue fibroblasts or another unexpected cell population. | Soft tissue fibroblasts are reported as not detected (HPA tissue IHC). Confirm cell identity and compare the nuclear pattern with a known positive section; cross-reactivity or endogenous detection activity may explain discordant staining (standard IHC practice). HPA's variable tissue expression limits blanket claims about every unlisted cell type. |
| Color spreads across stroma, empty spaces or multiple compartments without distinct nuclei. | That appearance is difficult to reconcile with predominantly nuclear RBM15 (HPA tissue IHC; UniProt Q96T37 subcellular location). Check the matched negative control and detection reagents for background (standard IHC practice). Background alone does not establish which reagent or step caused it. |
| There is no convincing nuclear signal in a known positive tissue. | A blank result in colon glands or kidney collecting ducts conflicts with their reported High staining (HPA tissue IHC). Check section quality, antibody identity and dilution, detection performance, and a positive control (standard IHC practice). A single blank slide cannot establish true absence of RBM15. |
💡Expected RBM15 appearanceCall a section positive when its expected cells show discernible, variable-intensity nuclear staining—for example, High staining in colon glands (HPA tissue IHC)—while diffuse cytoplasmic or cell-free color is treated as suspect (HPA tissue IHC; standard IHC practice).
| Compartment and resolution | UniProt lists nuclear speckles, nucleoplasm, envelope and membrane (UniProt Q96T37 subcellular location); HPA tissue IHC summarizes nuclear staining. Chromogenic sections may support a nuclear call without resolving each subnuclear site (standard IHC interpretation). Avoid requiring a sharply punctate pattern in every positive cell. |
| Cell population and tissue choice | Purkinje cells, intestinal glands, kidney collecting ducts and placental decidual cells are High; soft tissue fibroblasts are not detected (HPA tissue IHC). Thyroid, adrenal and salivary glandular cells are reported Low (HPA tissue IHC). Choose controls by the specified cell population, not the organ name alone. |
| Evidence strength | The overall tissue profile is Supported, with medium agreement between antibody staining and RNA expression (HPA tissue IHC). HPA019824 and CAB015201 have Supported IHC status; HPA049642 has no IHC status listed (HPA antibodies). These labels support cautious pattern interpretation, not universal antibody performance. |
| Isoforms and antibody epitope | UniProt lists 4 isoforms and RRM and SPOC domains (UniProt Q96T37). No epitope location is supplied here, so isoform coverage cannot be inferred for an individual antibody. Confirm the catalog antibody's documented target region before interpreting discordant staining as an isoform-specific pattern. |
| RNA enrichment versus protein staining | Bone marrow is tissue enhanced at the RNA level, while the supplied protein profile reports variable nuclear expression and named High cell populations elsewhere (HPA tissue IHC). RNA enrichment alone does not set an IHC intensity threshold for marrow (standard interpretation of RNA and protein measurements). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected nuclei are blank in a colon gland positive control. | The result conflicts with High glandular staining (HPA tissue IHC); the slide alone does not identify the failed step. | Verify antibody identity and documented IHC use, dilution, detection reagents and counterstain; repeat with a known positive section and controls (standard IHC practice). Record any antigen retrieval adjustment as a general workflow test, since RBM15-specific fixation sensitivity is unreported here. |
| Signal is broad and cytoplasmic in a tissue with visible nuclei. | Predominantly nuclear staining is expected (HPA tissue IHC; UniProt Q96T37 subcellular location); detection background or nonspecific binding is possible (standard IHC practice). | Compare a matched negative control, inspect the nuclear counterstain, and reassess blocking and detection conditions (standard IHC practice). Accept a nuclear rim only when it is clearly tied to nuclei (UniProt Q96T37 subcellular location). |
| Fibroblasts stain as strongly as the intended positive cells. | Soft tissue fibroblasts are reported not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible, but staining alone does not distinguish them (standard IHC practice). | Confirm morphology and compare no-primary and detection controls, then repeat with an independently validated IHC antibody if available (standard IHC practice; HPA antibodies: two Supported IHC entries). |
| A low-staining gland is scored negative beside a strong glandular control. | HPA reports Low staining in thyroid, adrenal and salivary glandular cells, but High staining in colon glands (HPA tissue IHC). | Score each named cell population against its own expected range and a same-run positive control (HPA tissue IHC; standard IHC practice). Do not apply the colon intensity threshold to a reported Low population. |
| Two antibodies give different nuclear staining patterns. | IHC status is antibody specific: HPA019824 and CAB015201 are Supported, while HPA049642 has no listed IHC status (HPA antibodies). Epitope and isoform coverage are not supplied here. | Check each antibody's documented IHC validation and epitope information, then compare the same positive and negative control sections (standard IHC practice). Treat disagreement as unresolved until the control patterns are interpretable. |
| IF/ICC Q: where should a corroborating cellular signal appear? | HPA reports enhanced nucleoplasmic localization in ICC-IF, consistent with a nuclear IHC call (HPA subcellular; HPA tissue IHC). | Look for nucleoplasmic signal aligned with nuclei, while interpreting the paraffin-section result against its own tissue controls (HPA subcellular; standard IHC interpretation). HPA's ICC-IF images include A-431, U-251MG, U2OS and NIH 3T3 (HPA subcellular). |
Experiment evidence
IHC & IF images showing RBM15 staining
A04395 · IHCIHC analysis of RBM15 using anti-RBM15 antibody (A04395). RBM15 was detected in a paraffin-embedded section of human breast cancer tissue. H
A04395 · IHCIHC analysis of RBM15 using anti-RBM15 antibody (A04395). RBM15 was detected in a paraffin-embedded section of human lung cancer tissue. Hea
A04395 · IFIF analysis of RBM15 using anti-RBM15 antibody (A04395) and anti-Tubulin Alpha antibody (M03989-3). RBM15 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162775-RBM15/tissue/cerebral+cortex#imid_5385820
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162775-RBM15/tissue/colon#imid_5385746
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162775-RBM15/tissue/liver#imid_5385739
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000162775-RBM15/tissue/kidney#imid_5385732
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB015201; green = RBM15, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB015201; green = RBM15, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RBM15 IHC & IF
🧪Run kidney first: collecting duct cells should show RBM15 staining (HPA: High in kidney collecting ducts). Use soft tissue fibroblasts as the negative tissue (HPA: Not detected in soft tissue fibroblasts); HPA does not identify a negative cell type within the kidney slide, so cells lacking nuclear signal should show counterstain only and serve as background, not a validated internal negative.
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Soft tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RBM15 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a concentration-matched rabbit immunoglobulin isotype control (selected-SKU tissue-IHC caption: rabbit anti-RBM15), and RBM15-knockout material as a biological negative. Block endogenous peroxidase for HRP/DAB detection and assess any residual tissue background on the no-primary slide (selected-SKU tissue-IHC caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected-SKU paraffin-section caption is unreported. That caption uses heat-mediated retrieval in EDTA at pH 8.0, so start there for IHC-P; it does not establish whether retrieval is required for every specimen (selected-SKU tissue-IHC caption). The supplied evidence does not show that frozen sections or IF are easier; kidney background should be checked with the no-primary control (HPA: High in kidney collecting ducts; selected-SKU tissue-IHC caption: HRP/DAB detection).
HPA tissue IHC evidence for RBM15
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Collecting ducts | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBM15 IHC Tips
Troubleshoot RBM15 staining by checking nuclear localisation, documented antibody conditions, cell type and controls before comparing sections.
Which retrieval conditions should I try first for RBM15 in paraffin sections?
Begin with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04395). The documented tissue image used this retrieval before 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A04395). If nuclear staining remains weak, compare a second retrieval condition on adjacent sections while keeping antibody concentration, detection and development constant (standard IHC practice). Examine tissue preservation and background alongside signal, because excessive heating can damage morphology or expose nonspecific binding (standard IHC practice). Score nuclei in the same cell population across conditions; RBM15 is reported in the nucleoplasm and nuclear speckles (HPA subcellular; UniProt Q96T37 localisation).
Could fixation explain weak RBM15 staining in paraffin sections?
Target-specific fixation sensitivity is unknown: the A04395 tissue caption reports a paraffin section but does not state its fixative (datasheet A04395). Record the fixative and processing history for every specimen before attributing a staining difference to RBM15 abundance (standard IHC practice). For a controlled comparison, process adjacent sections with the documented EDTA pH 8.0 retrieval and 2 μg/ml primary antibody overnight at 4°C (datasheet A04395). Keep section thickness, detection and chromogen development consistent, then assess nuclear signal and tissue morphology together (standard IHC practice). Neither a nuclear staining pattern nor the protein’s modifications establish a particular fixation requirement (HPA tissue IHC; UniProt Q96T37 modified residues).
Where should convincing RBM15 staining appear in chromogenic IHC?
Prioritise staining within intact nuclei: tissue IHC shows variable nuclear expression, while subcellular evidence emphasises the nucleoplasm (HPA tissue IHC; HPA subcellular). RBM15 is also annotated at nuclear speckles, the nuclear envelope and the nuclear membrane, with nuclear pore colocalisation reported (UniProt Q96T37 localisation). Its sequence has no transmembrane segment, so a dominant cell surface rim needs independent verification before assignment to RBM15 (UniProt Q96T37 topology; standard IHC practice). Compare nuclear signal with the counterstain and inspect several fields away from section edges (standard IHC practice). Use matched negative controls to assess whether diffuse cytoplasmic colour follows nonspecific detection rather than the expected nuclear distribution (standard IHC practice).
How might RBM15 isoforms or epitope location affect IHC results?
RBM15 has 4 annotated isoforms, but the supplied A04395 caption does not identify its epitope or establish isoform coverage (UniProt Q96T37 isoforms; datasheet A04395). The protein contains 3 RNA recognition motifs and a SPOC domain at residues 777–956 (UniProt Q96T37 domains). Check the catalog antibody’s immunogen or epitope documentation before interpreting a negative section as absence of every isoform (standard IHC practice). Retrieval may change access to a fixed epitope, so compare adjacent sections under controlled conditions if staining is discordant (standard IHC practice). Keep the documented EDTA pH 8.0 condition as the reference when evaluating those differences (datasheet A04395).
How should I assess RBM15 alongside cell markers by IF?
Use a cell-type marker appropriate to the compartment being examined; HPA reports high nuclear staining in several glandular cell populations (HPA tissue IHC). Choose spectrally separated fluorophores and consider a far-red RBM15 channel when tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because RBM15 is nuclear and lacks a transmembrane segment, assess nuclear access through permeabilisation rather than assuming a cell surface epitope (UniProt Q96T37 localisation and topology; standard IF practice). Include single-colour and secondary-only controls to evaluate bleed-through and background (standard IF practice). Treat IF settings as a separate optimisation: the A04395 caption documents paraffin-section chromogenic IHC, not IF fixation, permeabilisation or dilution (datasheet A04395).
What should I change when RBM15 DAB staining obscures nuclei?
First compare a no-primary control with the test section to locate background from secondary detection, endogenous peroxidase or chromogen development (standard IHC practice). The documented A04395 workflow used 10% goat serum block, 2 μg/ml primary antibody overnight at 4°C, peroxidase-linked detection and DAB (datasheet A04395). Include a peroxidase block and monitor DAB development as general chromogenic IHC steps; their effectiveness in a new specimen must be checked experimentally (standard IHC practice). If background persists, optimise primary concentration and washes while keeping retrieval at EDTA pH 8.0 initially (datasheet A04395; standard IHC practice). Judge improvement by nuclear contrast and preserved morphology, not by colour intensity alone (HPA tissue IHC; standard IHC practice).
How can I quantify RBM15 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and nuclear positivity threshold before comparing specimens (standard IHC practice). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score from 0–300 using intensity categories 0–3 (standard IHC practice). Normalise counts to the number of eligible intact nuclei within the same annotated tissue compartment, and report cells per mm² if density matters (standard IHC practice). Exclude necrotic areas, folds and damaged edges by a consistent rule (standard IHC practice). Keep retrieval, antibody concentration, DAB development and image acquisition comparable; HPA describes variable nuclear expression across tissues (datasheet A04395; HPA tissue IHC; standard IHC practice).
How do I distinguish genuine RBM15 staining from an IHC artefact?
A credible result is predominantly nuclear in intact cells, consistent with RBM15’s reported nucleoplasmic localisation and variable nuclear tissue staining (HPA subcellular; HPA tissue IHC). Check cell identity: HPA reports high staining in colon glandular cells but no detected staining in soft-tissue fibroblasts (HPA tissue IHC). Treat staining limited to section edges, folds or necrotic zones as suspect until it reproduces in well-preserved fields (standard IHC practice). Use no-primary and peroxidase-block controls to investigate diffuse DAB colour or endogenous enzyme activity (standard IHC practice). Interpret a negative specimen cautiously because HPA rates tissue staining as Supported with medium agreement between staining and RNA expression (HPA tissue IHC).
Boster reagents
Best RBM15 / RNA-binding protein 15 IHC Antibodies
A04395 has IHC images from paraffin sections of human breast and lung cancer and mouse and rat brain, plus IF images from U2OS cells and human lung cancer sections (A04395 image captions).
Real IHC data
Anti-RBM15 Antibody ®
Cat # A04395
A04395 is the only SKU shown; its IHC images cover paraffin sections of human breast and lung cancer and mouse and rat brain (A04395 IHC captions). Its IF images show U2OS cells and human lung cancer sections, and the catalog lists both IF and ICC applications (A04395 IF captions; catalog applications).
Which to pick: Choose A04395 for tissue IHC on paraffin sections; its own IHC captions show staining at 2 μg/ml in human, mouse and rat samples (A04395 IHC captions). For IF/ICC, A04395 lists both applications and has IF images from U2OS cells and human lung cancer sections (catalog applications; A04395 IF captions). For cross-species IHC, A04395 lists Human, Mouse and Rat reactivity and has IHC images for all three; clonality and the tissue fixative are unreported (catalog reactivity and clone field; A04395 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96T37 (RBM15_HUMAN, RNA-binding protein 15).
- Human Protein Atlas. RBM15 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RBM15 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RBM15 antibody validation summary (3 antibodies).
- RBM15 Promates the Proliferation, Migration and Invasion of Pancreatic Cancer Cell Lines. Cancers 2023 — PMC9954619.
- Effect of the m6ARNA gene on the prognosis of thyroid cancer, immune infiltration, and promising immunotherapy. Frontiers in immunology 2022 — PMC9664221.
- RBM15 recruits myeloid-derived suppressor cells via the m6A-IGF2BP3/CBR3-AS1/miR-409-3p/CXCL1 axis, facilitating radioresistance in non-small-cell lung cancer. Journal of translational medicine 2025 — PMC11831794.
- RBM15 suppresses hepatic insulin sensitivity of offspring of gestational diabetes mellitus mice via m6A-mediated regulation of CLDN4. Molecular medicine (Cambridge, Mass.) 2023 — PMC9942341.
- PubMed PMID:11431691 — UniProt-cited evidence.
- PubMed PMID:11344311 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RBM15 (RNA-binding protein 15). Expected localisation Variable nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04395). Validated IHC antibody: A04395 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Soft tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A04395); 2. Fixation: Image fixative and duration unreported (datasheet A04395); 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 A04395); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04395); 8. Primary antibody: Rabbit anti-RBM15, 2-5 μg/ml (datasheet A04395); 9. Primary incubation: Overnight at 4 °C (datasheet A04395). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04395); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: Four isoforms; check epitope coverage across variants (UniProt); Regulation: Variable by tissue; marrow RNA enhanced (HPA tissue RNA). Evidence: UniProt Q96T37, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A04395, PMC9664221, PMC11831794. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RBM15/immunohistochemistry-ihc-guide.