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RBM22 / Pre-mRNA-splicing factor RBM22 · IHC design guide
Design Immunohistochemistry for RBM22
Plan RBM22 IHC in paraffin sections with nuclear scoring and appendix lymphoid tissue as a high-staining reference (HPA tissue IHC). Start the catalog antibody at 2–5 μg/ml (datasheet A10311-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RBM22 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 | Predominantly nuclear in tissue (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across diverse tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10311-1) | |
| Positive control | Adrenal gland+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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10311-1) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Heat shock shifts RBM22 to cytoplasm (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended RBM22 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet: A10311-1); two published RBM22 IHC protocols provide additional conditions (PMC12205502; PMC7904102).
| Sample | Paraffin-embedded human appendix adenocarcinoma tissue; fixative not specified (datasheet A10311-1) |
| Fixation | Image fixative and duration unreported (datasheet A10311-1); 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 A10311-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10311-1) |
| Primary antibody | Rabbit anti-RBM22, 2-5 μg/ml (datasheet A10311-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10311-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10311-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RBM22-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A10311-1). The craniopharyngioma study lists alternative buffers without assigning one to RBM22 (PMC12205502).
Section 2
What Is the Expected RBM22 Staining Pattern?
RBM22 should stain predominantly in nuclei across many cell types in paraffin sections (HPA: ubiquitous nuclear expression; UniProt Q9NW64: nearly exclusively nuclear). HPA reports high staining in breast glandular cells, bone marrow hematopoietic cells, and several other listed populations (HPA: tissue IHC). Its tissue IHC reliability is Supported, with medium consistency between antibody staining and RNA expression (HPA: reliability). RBM22 has no transmembrane segment (UniProt Q9NW64: topology).
| Distinct nuclear chromogen in breast glandular cells or bone marrow hematopoietic cells, with readable nuclei and limited background. | This matches two reported high-staining populations and the reported ubiquitous nuclear profile (HPA: tissue IHC). Judge intensity within the stained cell population; HPA's High category does not prescribe a chromogen score or exposure setting (HPA: tissue IHC). |
| Predominantly cytoplasmic staining in an otherwise well-preserved, unstressed section, with little nuclear signal. | Treat this as discordant with the baseline pattern and investigate background or antibody specificity (HPA: nuclear IHC profile; UniProt Q9NW64: nearly exclusively nuclear). Cytoplasmic RBM22 is biologically possible after heat shock, so compartment alone cannot prove an artefact (UniProt Q9NW64: heat-shock translocation). |
| Strong stain confined to a cell population expected to show low staining, while nearby expected nuclei remain faint. | Check cell identification and controls before calling cross-reactivity or endogenous detection activity (HPA: low in liver cholangiocytes, parathyroid glandular cells, and ovarian stroma cells). Low is not negative, and HPA describes broad nuclear expression (HPA: tissue IHC). |
| Diffuse colour across nuclei, cytoplasm, and tissue spaces, with little distinction between cells. | This cannot establish the expected nuclear pattern (HPA: ubiquitous nuclear IHC profile). Review antibody concentration, blocking, washes, and detection controls as general chromogenic IHC practice; the supplied sources do not identify which workflow step caused the background. |
| No nuclear staining in an interpretable breast or bone marrow positive-control section. | The result conflicts with reported High staining in those populations (HPA: breast glandular cells; bone marrow hematopoietic cells). Check section quality and the staining run before interpreting a study specimen as RBM22-negative (general IHC practice). HPA's Supported rating is not a guarantee for every section (HPA: reliability). |
💡Expected RBM22 appearanceCall a result consistent with RBM22 when chromogen is mainly nuclear in identifiable cells, including strong staining in HPA-listed High populations; isolated cytoplasmic colour or diffuse stain without nuclear contrast warrants scrutiny (HPA: tissue IHC; UniProt Q9NW64: nearly exclusively nuclear).
| Cellular compartment | Nuclear localisation is the primary interpretation anchor (HPA: ubiquitous nuclear IHC expression; UniProt Q9NW64: nearly exclusively nuclear). Heat shock can move RBM22 toward the cytoplasm, so document stress context before treating cytoplasmic staining as biological (UniProt Q9NW64: heat-shock translocation). |
| Tissue and cell population | HPA reports High staining in listed glandular, hematopoietic, respiratory epithelial, neuronal, and glial populations, and Low staining in three listed populations (HPA: tissue IHC). These categories guide control selection and comparisons within identified cell types; they do not define an unstained tissue (HPA: no negative category listed). |
| Antibody evidence | The listed antibody HPA001634 has Supported IHC and Enhanced ICC validation (HPA: antibody validation). HPA describes tissue IHC as having medium consistency with RNA expression (HPA: reliability). These ratings support pattern interpretation but do not establish specificity for a different antibody or staining run. |
| ICC-IF comparison | HPA reports enhanced nucleoplasmic localisation in ICC-IF images (HPA: subcellular ICC-IF). This independently supports a nuclear expectation, while ICC-IF images cannot set chromogenic intensity thresholds for paraffin sections (HPA: subcellular ICC-IF; general IHC interpretation). IF/ICC workflow belongs in its own guide. |
| Topology and sequence features | RBM22 lacks a transmembrane segment and has two annotated isoforms (UniProt Q9NW64: topology; isoforms). These facts fit an intracellular interpretation, but they do not identify the catalog antibody's epitope or predict isoform-specific staining. The supplied sources give no target-specific fixation-sensitivity result. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control nuclei are blank. | A staining-run or section problem is possible; the observation conflicts with reported High control populations (HPA: breast glandular cells; bone marrow hematopoietic cells). | Verify tissue identification and preservation, then review retrieval, primary-antibody application, detection reagents, and counterstain on the same run (general IHC practice). Do not label a study specimen negative until the control is interpretable. |
| Most colour is cytoplasmic. | The distribution differs from baseline nuclear localisation (HPA: tissue IHC; UniProt Q9NW64: nearly exclusively nuclear). Heat-shock translocation remains a contextual possibility (UniProt Q9NW64: heat-shock translocation). | Confirm whether the sample underwent relevant stress; inspect morphology and detection controls, then repeat with an appropriate positive control if the compartment remains uncertain (general IHC practice). |
| Low-staining populations appear as intense as listed High populations. | Cell identification, background, or nonspecific detection may affect the comparison (HPA: Low in cholangiocytes, parathyroid glandular cells, and ovarian stroma cells; High in listed populations). | Compare the same run at matched staining conditions, verify cell identity, and inspect a no-primary control for detection-related colour (general IHC practice). Do not interpret Low as absent (HPA: tissue IHC). |
| Brown haze obscures nuclear boundaries. | Diffuse background prevents a reliable compartment call (HPA: nuclear IHC profile); its source cannot be assigned from the supplied target evidence. | Check blocking, primary concentration, washes, detection timing, and a no-primary control according to the staining workflow (general IHC practice). Reassess only when individual nuclei are readable. |
| Scattered isolated cells stain strongly in an otherwise weak section. | RBM22 staining spans multiple cell populations, so isolated intensity alone cannot identify an unexpected cell type or establish cross-reactivity (HPA: ubiquitous nuclear IHC expression; listed High and Low populations). | Identify the stained cells morphologically, compare their nuclear and cytoplasmic compartments, and check adjacent cells and controls before assigning biological meaning (general IHC interpretation). |
| IHC and ICC-IF appear to disagree. | HPA reports nuclear tissue IHC and enhanced nucleoplasmic ICC-IF, but the methods and specimens differ (HPA: tissue IHC; subcellular ICC-IF). Their signal intensities are not directly comparable. | Compare localisation within each method, document sample stress, and resolve discrepancies with method-appropriate controls; use the separate IF/ICC guide for its workflow (UniProt Q9NW64: heat-shock translocation; general IHC practice). |
Experiment evidence
IHC & IF images showing RBM22 staining
A10311-1 · IHCIHC analysis of RBM22 using anti-RBM22 antibody (A10311-1). RBM22 was detected in a paraffin-embedded section of human appendix adenocarcino
A10311-1 · IHCIHC analysis of RBM22 using anti-RBM22 antibody (A10311-1). RBM22 was detected in a paraffin-embedded section of human breast cancer tissue.
A10311-1 · IFIF analysis of RBM22 using anti-RBM22 antibody (A10311-1). RBM22 was detected in a paraffin-embedded section of mouse brain tissue. Heat med
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000086589-RBM22/tissue/cerebral+cortex#imid_1120944
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000086589-RBM22/tissue/colon#imid_1120870
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000086589-RBM22/tissue/liver#imid_1120871
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000086589-RBM22/tissue/kidney#imid_1120880
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001634; green = RBM22, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001634; green = RBM22, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RBM22 IHC & IF
🧪Run appendix first and assess nuclear staining in its lymphoid tissue (HPA: appendix lymphoid tissue, High; HPA subcellular: nucleoplasm enhanced). HPA lists no negative tissue because RBM22 is detected in all 45 scored tissues; no cell population on the positive slide is established as RBM22-negative, so use no-primary and isotype sections to confirm that counterstained nuclei lack specific DAB signal (HPA: no negative rows; selected-SKU caption: HRP/DAB).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: RBM22 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 RBM22 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, siRNA 2 (10x), U2OS, siRNA 2 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only section and a matched rabbit IgG isotype section; use RBM22 knockout material or cognate-peptide competition, if available, as a biological specificity control (selected-SKU caption: rabbit primary and goat anti-rabbit secondary). Quench endogenous peroxidase in appendix sections before HRP/DAB detection, particularly where myeloid cells are present (HPA: appendix lymphoid tissue; selected-SKU caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A10311-1 paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA pH 8.0, but whether retrieval is required is unreported (selected-SKU caption: EDTA pH 8.0). Frozen-section performance is unreported; ICC-IF images show enhanced nucleoplasmic localization without establishing that IF is easier, while endogenous peroxidase in appendix myeloid cells could confound chromogenic scoring (HPA subcellular: nucleoplasm enhanced; HPA: appendix lymphoid tissue; selected-SKU caption: HRP/DAB).
HPA tissue IHC evidence for RBM22
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RBM22 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RBM22 IHC Tips
Use nuclear staining as the main readout for RBM22 in chromogenic paraffin section IHC, with attention to retrieval and tissue background (HPA tissue IHC; UniProt Q9NW64).
What retrieval should I try when RBM22 nuclear staining is weak?
Start with heat mediated antigen retrieval in EDTA buffer at pH 8.0 (datasheet A10311-1). The catalog antibody stained a paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A10311-1). If nuclei remain faint, check that sections reached the intended retrieval temperature and cooled consistently before changing chemistry (standard IHC practice). Compare any longer retrieval with the original conditions on adjacent sections, since excessive heating can damage morphology and complicate nuclear scoring (standard IHC practice). Keep detection and counterstaining matched so a retrieval change is the main variable (standard IHC practice).
Could fixation explain inconsistent RBM22 staining between paraffin blocks?
The selected image identifies a paraffin section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A10311-1). Record each block’s fixative and fixation time, then compare sections processed with the same retrieval and detection conditions (standard IHC practice). Include a reference block with reproducible nuclear staining in each run; widespread loss in that block points toward a run problem rather than specimen variation (standard IHC practice; HPA tissue IHC). If blocks differ in fixation history, assess morphology and staining together before changing EDTA pH 8.0 retrieval (datasheet A10311-1; standard IHC practice). Do not assign a fixation mechanism from the observed staining pattern alone (standard IHC practice).
How should I assess cytoplasmic RBM22 staining in an otherwise nuclear section?
Score nuclear staining first: RBM22 is nearly exclusively nuclear under usual conditions, and tissue IHC shows ubiquitous nuclear expression (UniProt Q9NW64 subcellular location; HPA tissue IHC). Cytoplasmic signal can be biologically plausible after heat shock, but that observation does not establish the state of a tissue section (UniProt Q9NW64 subcellular location). Check whether the cytoplasmic color is confined to intact cells or also covers stroma, damaged areas, and section edges (standard IHC practice). Review a matched no primary control for chromogen deposition, and compare the nuclear pattern with a reference section (standard IHC practice). Report nuclear and cytoplasmic scores separately when both persist (standard IHC practice).
Can this antibody distinguish RBM22 isoforms or detect a modified epitope?
RBM22 has 2 annotated isoforms, but the supplied IHC caption does not map the catalog antibody’s epitope or establish isoform specificity (UniProt Q9NW64 isoforms; datasheet A10311-1). Its RRM spans residues 232–305, while annotated modifications include phosphoserines at residues 4 and 102 and acetyllysine at 212 (UniProt Q9NW64 domains and modified residues). Those annotations alone cannot predict whether retrieval exposes or masks this antibody’s epitope (standard IHC practice). Compare nuclear staining with an independently validated antibody to a documented, distinct epitope if isoform or modification dependence matters (standard IHC practice). Describe the resulting stain as RBM22 immunoreactivity until specificity for the proposed form is established (standard IHC practice).
How can I check RBM22 localisation with multiplex IF?
Use IF as a separate localisation check, pairing RBM22 with a marker for the expected cell type and a nuclear counterstain (standard IF practice). Assign well separated fluorophores and place the weaker channel away from the tissue’s strongest autofluorescence; inspect single channel controls before judging overlap (standard IF practice). RBM22 is nearly exclusively nuclear, and HPA reports enhanced nucleoplasmic localisation, so confirm that permeabilisation allows antibody access to the nuclear epitope (UniProt Q9NW64 subcellular location; HPA subcellular; standard IF practice). The protein has no transmembrane segment, but its exact antibody epitope is unreported here (UniProt Q9NW64 topology; datasheet A10311-1). Optimise IF fixation and permeabilisation independently; the paraffin IHC caption does not establish IF conditions (datasheet A10311-1; standard IF practice).
How do I separate genuine nuclear staining from DAB background?
First inspect a no primary control processed through the same peroxidase detection and DAB development, since deposits there indicate assay background (standard IHC practice). The selected image used 10% goat serum blocking, a peroxidase conjugated secondary, and DAB, but it does not document a peroxidase blocking step (datasheet A10311-1). Add a peroxidase block as a general chromogenic IHC control and check whether brown signal outside nuclei decreases (standard IHC practice). Compare intact nuclei with section edges and damaged regions, where uneven reagent access or nonspecific deposits can mislead scoring (standard IHC practice). Preserve the same DAB development time across comparison slides (standard IHC practice).
What is a defensible way to quantify RBM22 tissue IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear RBM22 separately from any cytoplasmic signal, using the same compartment rule for every section (HPA tissue IHC; UniProt Q9NW64 subcellular location; standard IHC practice). For an H-score, record the percentage of cells at each staining intensity and calculate 0 × % negative + 1 × % weak + 2 × % moderate + 3 × % strong (standard IHC practice). Alternatively, report the percentage of positive nuclei with a prespecified threshold rather than counting DAB area alone (standard IHC practice). Normalise counts to evaluable cells in the defined tissue region and exclude folds, necrosis, and edge artefacts (standard IHC practice). Keep retrieval, exposure, and scoring thresholds consistent across slides (standard IHC practice).
When should an apparent RBM22 positive result be treated cautiously?
A credible result shows staining in intact nuclei within the scored cell population; HPA describes ubiquitous nuclear expression with supported reliability and medium RNA concordance (HPA tissue IHC). Strong staining confined to cytoplasm, section edges, or necrotic material warrants review before calling it RBM22 positive (UniProt Q9NW64 subcellular location; standard IHC practice). Check the no primary control and peroxidase block if brown deposits appear in the same pattern without primary antibody (standard IHC practice). HPA reports high staining in appendix lymphoid tissue and lower staining in liver cholangiocytes, so compare like cell populations when interpreting intensity (HPA tissue IHC). Atypical cytoplasmic staining needs independent support before attributing it to stress related translocation (UniProt Q9NW64 subcellular location; standard IHC practice).
Boster reagents
Best RBM22 / Pre-mRNA-splicing factor RBM22 IHC Antibodies
A10311-1 has IHC images from human paraffin sections and IF images from mouse and rat paraffin brain sections (catalog image captions). Its listed reactivity covers human, mouse, and rat (catalog applications/reactivity).
Real IHC data
Anti-RBM22 Antibody ®
Cat # A10311-1
A10311-1 has IHC images from human appendix adenocarcinoma, breast cancer, esophageal squamous carcinoma, and gastric adenocarcinoma paraffin sections (A10311-1 IHC captions). Its IF images show mouse and rat brain paraffin sections (A10311-1 IF captions).
Which to pick: Choose A10311-1 for tissue IHC: its human paraffin-section captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (A10311-1 IHC captions). For IF, A10311-1 has mouse and rat tissue images at 5 μg/ml, while ICC validation is unreported (A10311-1 IF captions; catalog applications). For cross-species work, A10311-1 lists human, mouse, and rat reactivity, although its IHC images document human tissue only (catalog reactivity; A10311-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NW64 (RBM22_HUMAN, Pre-mRNA-splicing factor RBM22).
- Human Protein Atlas. RBM22 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RBM22 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RBM22 antibody validation summary (1 antibodies).
- Impaired splicing machinery in craniopharyngiomas unveils PRPF8 and RAVER1 as novel biomarkers and therapeutic targets. Acta neuropathologica communications 2025 — PMC12205502.
- Splicing machinery dysregulation drives glioblastoma development/aggressiveness: oncogenic role of SRSF3. Brain : a journal of neurology 2020 — PMC7904102.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RBM22 (Pre-mRNA-splicing factor RBM22). Expected localisation Predominantly nuclear in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10311-1). Validated IHC antibody: A10311-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (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 appendix adenocarcinoma tissue; fixative not specified (datasheet A10311-1); 2. Fixation: Image fixative and duration unreported (datasheet A10311-1); 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 A10311-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10311-1); 8. Primary antibody: Rabbit anti-RBM22, 2-5 μg/ml (datasheet A10311-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10311-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A10311-1); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage (UniProt); Regulation: Heat shock shifts RBM22 to cytoplasm (UniProt). Evidence: UniProt Q9NW64, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A10311-1, PMC12205502, PMC7904102. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RBM22/immunohistochemistry-ihc-guide.