This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
RBMX / RNA-binding motif protein, X chromosome · IHC design guide
Design Immunohistochemistry for RBMX
Plan RBMX paraffin-section IHC around its nuclear staining across most tissues (HPA tissue IHC). Use high-staining adrenal glandular cells and low-staining adipocytes as reference patterns, while accounting for the atlas warning that staining may detect products from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RBMX 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04738) | |
| 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 | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may detect products from more than one gene (HPA tissue IHC) | |
| Regulation | Weaker in oral squamous carcinomas (UniProt) | |
| Isoform / epitope | 3 isoforms; N-terminal processing may affect epitopes near residue 1 (UniProt) |
Section 1
Recommended RBMX IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published chromogenic RBMX IHC methods for lymphoma, esophageal carcinoma, and paraffin-embedded tissue (PMC8507920; PMC10583014; PMC10442647).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A04738) |
| Fixation | Image fixative and duration unreported (datasheet A04738); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A04738); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-RBMX, 1:100-1:300 (datasheet A04738) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RBMX-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA at pH 9.0, as specified for the catalog antibody and reported in one published method (datasheet A04738; PMC10442647).
Section 2
What Is the Expected RBMX Staining Pattern?
RBMX is nuclear, with numerous small nuclear granules, and has no transmembrane segment (UniProt P38159). In paraffin-section IHC, expect nuclear staining across many tissues, including respiratory epithelial, glandular and neuronal cells (HPA: ubiquitous nuclear profile; High in listed cells). HPA rates tissue IHC Supported, with medium agreement with RNA data, pending external verification, and cautions that staining may include proteins from more than one gene (HPA: reliability description).
| Clear nuclear staining in bronchial respiratory epithelial cells, with nuclei identifiable by counterstain. | This fits the expected compartment and a documented High-staining cell type (UniProt P38159: nucleus; HPA: High in bronchial respiratory epithelial cells). Record the proportion and intensity of stained nuclei while keeping tissue structure visible. This supports an interpretable staining pattern, although HPA cautions that the antibody evidence may include proteins from more than one gene (HPA: tissue IHC reliability description). |
| Predominantly cytoplasmic staining, with little or no nuclear signal in cells expected to stain. | Treat this as a compartment mismatch and investigate an artefact before assigning it to RBMX (UniProt P38159: nuclear localization; HPA: ubiquitous nuclear IHC profile). UniProt also describes a role in cytoplasmic TNFR1 trafficking, but that functional annotation does not establish predominantly cytoplasmic IHC staining (UniProt P38159: function). Check the counterstain and detection controls, then compare nuclear and cytoplasmic signal in the same cell type (general IHC practice). |
| Strong signal is confined to an unexpected cell population while documented High-staining cells lack nuclear signal. | Compare the actual cell identities with the tissue-specific HPA entry before calling the pattern unexpected: HPA reports High staining in colon endothelial cells, for example, and describes expression as broadly nuclear (HPA: colon entry; tissue profile). A persistent mismatch can indicate cross-reactivity or endogenous detection activity; morphology alone cannot distinguish those causes (general IHC practice). HPA's multiple-gene caution makes target attribution especially qualified (HPA: reliability description). |
| Diffuse colour across nuclei, cytoplasm and tissue spaces obscures cell boundaries. | This is difficult to score as RBMX because the supported pattern is nuclear (HPA: tissue IHC profile; UniProt P38159: localization). Assess whether background also appears in a no-primary control, and review blocking, washing and chromogen development as general IHC checks (general IHC practice). Diffuse background does not establish that RBMX is absent from the section. |
| No discernible nuclear signal in bronchial respiratory epithelium or another documented High-staining population. | Do not immediately interpret this as biological absence: these populations are reference points for an expected positive pattern (HPA: High in bronchial respiratory epithelial cells and other listed cells). Confirm that the expected cells are present, examine a concurrently stained positive section, and review the IHC-validated antibody and detection workflow (general IHC practice). HPA's Supported rating describes agreement, not a guarantee that every specimen will stain (HPA: reliability description). |
💡Expected RBMX appearanceCall the result pattern-consistent when identifiable nuclei in documented High-staining cells show strong, localized signal; predominantly cytoplasmic or diffuse signal is suspect (HPA: High tissue entries and nuclear profile; UniProt P38159: nucleus).
| Tissue and cell selection | Bronchial respiratory epithelial, cerebral cortical neuronal and adrenal glandular cells are documented High-staining examples; adipocytes are listed Low (HPA: tissue IHC entries). Use cell identity when comparing sections: a weak adipocyte signal is less informative than absent nuclear signal in a documented High-staining population (HPA: tissue IHC entries). |
| Antibody interpretation | The listed antibody HPA057707 has Supported IHC status, while HPA reports medium staining-to-RNA consistency, pending external verification, and possible detection of proteins from multiple genes (HPA: antibody status; reliability description). A plausible nuclear pattern alone therefore cannot prove RBMX-specific recognition. |
| Isoforms and processing | UniProt lists 3 isoforms and chains beginning at residue 1 or 2 (UniProt P38159: isoforms; processing). No antibody epitope is supplied, so the record cannot establish which isoforms or processed forms this IHC stain detects; avoid assigning a difference in intensity to one form. |
| IF/ICC Q&A: where should signal appear? | In IF/ICC, expect nucleoplasmic signal; UniProt also describes numerous small nuclear granules (HPA: supported nucleoplasm; UniProt P38159: localization). HPA lists HEK293, MCF-7 and U2OS image sets but cautions that its subcellular assignment uses antibodies targeting proteins from multiple genes (HPA: subcellular record). |
| Situation | Likely cause | Next action |
|---|---|---|
| A documented High-staining cell type has no nuclear signal. | The expected cells may be absent from the section, or the staining run may have failed; this observation alone cannot distinguish either possibility from biological variation (HPA: High tissue entries; general IHC practice). | Locate the expected cells by morphology and counterstain, then compare a concurrently processed positive section. Review antibody use and detection against the available IHC protocol before interpreting the result (general IHC practice). |
| Signal appears mainly outside nuclei. | The compartment conflicts with the supported nuclear pattern; non-specific staining or a detection artefact is possible (HPA: ubiquitous nuclear IHC profile; UniProt P38159: nucleus; general IHC practice). | Compare signal with the counterstained nuclei and check a no-primary control. If extranuclear colour persists, report the compartment mismatch rather than scoring it as the expected RBMX pattern (general IHC practice). |
| Broad tissue colour makes positive nuclei hard to identify. | Background from the staining or detection workflow may obscure localized signal; the record provides no RBMX-specific cause for diffuse background (general IHC practice). | Check a no-primary control and review blocking, washing and development time. Score nuclear staining only where nuclei and cell boundaries remain interpretable (general IHC practice). |
| Unexpected cells stain more strongly than the documented reference population. | Cell identification, cross-reactivity or endogenous detection activity could contribute; HPA also cautions about proteins from multiple genes (general IHC practice; HPA: reliability description). | Verify cell identity on the section, consult the matching HPA tissue entry, and inspect a detection control. Describe the observed cells and compartment without assigning target specificity from intensity alone (HPA: tissue IHC entries; general IHC practice). |
| Adipocytes stain weakly while nearby cells show nuclear signal. | Low adipocyte staining is documented and need not indicate a failed run (HPA: Low in adipocytes; ubiquitous nuclear tissue profile). | Judge run performance against a documented High-staining cell population on a suitable section. Record adipocyte staining separately by cell type and intensity (HPA: tissue IHC entries; general IHC practice). |
| A nuclear pattern looks convincing, but RBMX specificity is the deciding question. | Correct localization supports pattern agreement but does not resolve HPA's warning about detection of proteins from more than one gene (UniProt P38159: nucleus; HPA: reliability description). | Report the result as pattern-consistent and retain the specificity caveat. If a target-specific conclusion is required, seek an independent validation approach appropriate to the experiment (general IHC practice; HPA: reliability description). |
Experiment evidence
IHC & IF images showing RBMX staining
A04738 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA, pH9.0 was used for antigen retrieval. 2 Antibody was diluted a
A04738 · IFImmunofluorescence analysis of HepG2 cells, using hnRNP G Antibody. The picture on the right is blocked with the synthesized peptide.
A04738-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Tris-EDTA,pH9.0 was used for antigen retrieval. 2 Antibody was diluted at
A04738-1 · IFImmunofluorescence analysis of HepG2 cells, using hnRNP G Antibody. The picture on the right is blocked with the synthesized peptide.
M04738 · IHCM04738 staining RBMX in human testis tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Ti
A04738-2 · IHCImmunohistochemistry (IHC) analyzes of hnRNP G (M40) pAb in paraffin-embedded human brain tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000147274-RBMX/tissue/cerebral+cortex#imid_16819731
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000147274-RBMX/tissue/colon#imid_16820386
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000147274-RBMX/tissue/liver#imid_16820387
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000147274-RBMX/tissue/pancreas#imid_16820338
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA057707; green = RBMX, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA057707; green = RBMX, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RBMX IHC & IF
🧪Run bronchus first: respiratory epithelial cells should show strong RBMX staining (HPA: High in bronchus respiratory epithelial cells). HPA detects RBMX in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and do not designate other cells on the positive slide as true negatives—judge their staining against control background and the expected nuclear localization (HPA: no negative tissue rows; UniProt P38159: nucleus).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: RBMX 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 RBMX in HEK293, MCF-7, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a host- and immunoglobulin-class-matched isotype control for a monoclonal primary or host-matched nonimmune IgG for a polyclonal primary, and an RBMX-knockout biological negative (standard IHC controls). Quench endogenous peroxidase for chromogenic detection; if using avidin–biotin detection, check endogenous biotin in the bronchus section (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A04738 paraffin-section caption leaves the fixative unreported. That caption uses Tris–EDTA retrieval at pH 9.0 and a 1:200 primary dilution overnight at 4 °C, but it does not establish that retrieval is required for every specimen (selected-SKU IHC caption). The evidence does not establish whether frozen sections or IF are easier; for IF, expect nucleoplasmic staining and assess tissue autofluorescence against a no-primary control (HPA: nucleoplasm supported; standard IF practice).
HPA tissue IHC evidence for RBMX
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Pending external verification. Caution, targets protein from more than one gene.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RBMX is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RBMX IHC Tips
Use nuclear staining as the main readout for RBMX in paraffin section IHC, while checking controls and tissue context before interpreting intensity (UniProt P38159; HPA tissue IHC).
What retrieval conditions should I start with for RBMX paraffin section IHC?
Use heat-mediated Tris-EDTA at pH 9.0 for antigen retrieval (datasheet A04738). The selected paraffin-embedded tonsil image pairs that retrieval with a 1:200 primary antibody dilution incubated at 4°C overnight (caption A04738). Keep section thickness, heating, cooling, and detection conditions consistent while comparing retrieval runs, because each can change apparent nuclear staining (standard IHC practice). If staining remains weak, test a milder retrieval condition alongside the documented condition, with matched control sections, and record any gain in nuclear signal or background (standard IHC practice; UniProt P38159 localisation). Do not treat the tonsil caption as fixation evidence: its fixative is unreported (caption A04738).
How should I troubleshoot weak RBMX staining when fixation conditions vary?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the selected tonsil caption reports paraffin embedding but gives no fixative (caption A04738). Record the fixative, fixation duration, processing history, and section age for each specimen before comparing staining intensity (standard IHC practice). Run sections with differing histories in the same staining batch, using heat-mediated Tris-EDTA at pH 9.0 as the documented retrieval condition (datasheet A04738; standard IHC practice). Compare nuclear signal with morphology and background, then adjust one processing variable at a time if controls underperform (UniProt P38159 localisation; standard IHC practice). HPA tissue patterns do not establish a fixation effect for this target (HPA tissue IHC).
Which staining pattern is expected, and how should cytoplasmic signal be assessed?
Expect predominantly nuclear RBMX staining: UniProt describes numerous small nuclear granules, while HPA reports supported nucleoplasmic localisation (UniProt P38159; HPA subcellular). Review nuclear signal against a counterstain before scoring, and focus on intact cells with preserved morphology (standard IHC practice). RBMX has no transmembrane segment, so membrane outlines are not its expected primary staining pattern (UniProt P38159 topology and localisation). Its reported role in cytoplasmic TNFR1 trafficking warrants investigation of reproducible cytoplasmic staining, but does not establish a diagnostic cytoplasmic IHC pattern (UniProt P38159 function). Compare unexpected signal with a no-primary control and a section showing clear nuclear staining in the same run (standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent RBMX nuclear staining?
RBMX has 3 listed isoforms, an RRM spanning residues 8–86, and recorded modified residues including phosphoserines at 88 and 91 (UniProt P38159). The supplied caption does not identify the antibody epitope, so isoform coverage and sensitivity to those modifications cannot be assigned to this antibody (caption A04738; UniProt P38159). Check the antibody’s documented immunogen or epitope before attributing discordant staining to an isoform (standard antibody validation practice). If staining differs across specimens, first compare retrieval, section processing, and intact nuclear morphology under matched conditions (standard IHC practice; UniProt P38159 localisation). Use an independent epitope or orthogonal assay when a specific isoform claim matters (standard antibody validation practice).
How can I assess RBMX localisation by IF alongside a cell identity marker?
For IF, pair RBMX with a marker identifying the cell population under examination; HPA reports high staining in bronchial respiratory epithelial cells and several glandular cell populations (HPA tissue IHC). Select spectrally distinct fluorophores and place the weaker signal in a channel with less tissue autofluorescence, checking single-stain controls for bleed-through (standard IF practice). Because the expected RBMX signal is nucleoplasmic and RBMX has no transmembrane segment, use permeabilisation that allows antibody access across the plasma and nuclear membranes (HPA subcellular; UniProt P38159 topology; standard IF practice). Include a nuclear counterstain to judge compartment overlap (standard IF practice). Validate IF fixation and antibody performance independently: the selected paraffin section caption supplies no IF fixation conditions (caption A04738).
What should I check when RBMX chromogenic IHC shows diffuse or patchy background?
Start by distinguishing crisp nuclear staining from diffuse chromogen in stroma, damaged areas, or section edges (UniProt P38159 localisation; standard IHC practice). Check a no-primary control and review blocking, washing, antibody dilution, and detection exposure in the same staining run (standard IHC practice). The selected image used 1:200 primary antibody overnight at 4°C and 1:200 secondary antibody for 45 minutes at room temperature; these are documented image conditions, not a background threshold (caption A04738). For peroxidase detection, verify the routine endogenous peroxidase block and inspect the control before attributing brown deposits to RBMX (standard chromogenic IHC practice). Compare suspect staining with counterstained nuclear morphology (standard IHC practice; UniProt P38159 localisation).
How should I quantify RBMX staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define a nuclear scoring rule before reviewing groups, because nuclear localisation is the primary expected pattern (UniProt P38159; HPA subcellular). Record either the percentage of positive intact nuclei and their intensity as an H-score, or the density of positive nuclei per mm² in a defined tissue compartment (standard IHC quantification practice). Normalise positive counts to all evaluable nuclei in that compartment, or report area-normalised density with the sampled area stated (standard IHC quantification practice). Apply the same threshold, counterstain review, and region selection to every section, excluding folds, edges, and necrotic areas (standard IHC quantification practice). Report cell population and staining batch alongside the score because HPA describes widespread nuclear expression (HPA tissue IHC).
When is an RBMX-positive IHC result convincing rather than artefactual?
A convincing result has reproducible staining in intact nuclei with a clear counterstain-defined boundary, consistent with UniProt nuclear localisation and HPA’s nucleoplasmic pattern (UniProt P38159; HPA subcellular; standard IHC practice). Interpret the stained cell population in context: HPA reports widespread nuclear expression and high staining in several specified epithelial, glandular, neuronal, and endothelial populations (HPA tissue IHC). Treat edge-restricted deposits, necrotic areas, and staining that also appears in a no-primary control as suspect (standard IHC practice). Investigate strong membrane-only staining because RBMX has no transmembrane segment and its principal reported location is nuclear (UniProt P38159 topology and localisation). Exclude endogenous peroxidase signal with the appropriate detection control before calling a brown reaction positive (standard chromogenic IHC practice).
Boster reagents
Best RBMX / RNA-binding motif protein, X chromosome IHC Antibodies
The catalog includes RBMX antibodies with human tissue IHC images for tonsil, brain and testis, plus IF images in HepG2 cells (catalog image captions). Human, mouse and rat reactivity is listed for selected SKUs (catalog reactivity).
Real IHC data
Anti-hnRNP G RBMX Antibody
Cat # A04738
Real IHC data
Anti-hnRNP G RBMX Antibody
Cat # A04738-1
Real IHC data
Anti-RBMX Antibody (Center)
Cat # M04738
Real IHC data
Anti-hnRNP G (M40) RBMX Antibody
Cat # A04738-2
A04738 and A04738-1 each show IHC in paraffin-embedded human tonsil and IF in HepG2 cells (catalog image captions); A04738-2 shows IHC in paraffin-embedded human brain (catalog IHC image caption). M04738 shows IHC in formaldehyde-fixed, paraffin-embedded human testis (catalog IHC image caption), and its application list includes IHC-P but excludes IF/ICC (catalog applications).
Which to pick: For tissue IHC, choose A04738 or A04738-1 for paraffin-embedded tonsil examples, A04738-2 for a paraffin-embedded brain example, or M04738 for a paraffin-section, paraffin-embedded testis example (each SKU’s IHC image caption); the fixative is unreported for A04738, A04738-1 and A04738-2 (their IHC image captions). For IF/ICC, A04738-1 lists both applications, while A04738 lists IF; both have HepG2 IF images and are listed as polyclonal (catalog applications, IF image captions and dilution data). For work across human, mouse and rat samples, consider A04738, A04738-1 or A04738-2 based on listed reactivity, while treating their tissue IHC images as human examples (catalog reactivity and IHC image captions). The selected A04738 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A04738).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P38159 (RBMX_HUMAN, RNA-binding motif protein, X chromosome).
- Human Protein Atlas. RBMX tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RBMX subcellular location (ICC-IF): Localized to the nucleoplasm. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. RBMX antibody validation summary (1 antibodies).
- RBMX Protein Expression in T-Cell Lymphomas Predicts Chemotherapy Response and Prognosis. Cancers 2021 — PMC8507920.
- Aberrant RBMX expression is relevant for cancer prognosis and immunotherapy response. Aging 2024 — PMC10817375.
- m6A reading protein RBMX as a biomarker for prognosis and tumor progression in esophageal cancer. Translational cancer research 2023 — PMC10583014.
- Identification of a novel lncRNA prognostic signature and analysis of functional lncRNA AC115619.1 in hepatocellular carcinoma. Frontiers in pharmacology 2023 — PMC10442647.
- PubMed PMID:7692398 — UniProt-cited evidence.
- PubMed PMID:11420617 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RBMX (RNA-binding motif protein, X chromosome). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A04738). Validated IHC antibody: A04738 at 1:100-1:300 (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 workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A04738); 2. Fixation: Image fixative and duration unreported (datasheet A04738); 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: Tris-EDTA pH 9.0 (datasheet A04738); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-RBMX, 1:100-1:300 (datasheet A04738); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining may detect products from more than one gene (HPA tissue IHC); Isoform / epitope: 3 isoforms; N-terminal processing may affect epitopes near residue 1 (UniProt); Regulation: Weaker in oral squamous carcinomas (UniProt). Evidence: UniProt P38159, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm. Caution: Based on antibodies targeting proteins from…, Datasheet A04738, PMC8507920, PMC10583014. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RBMX/immunohistochemistry-ihc-guide.