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RBM3 / RNA-binding protein 3 · IHC design guide
Design Immunohistochemistry for RBM3
Plan RBM3 paraffin section IHC around the nuclear staining seen in most tissues (HPA tissue IHC). Use nasopharyngeal basal cells as a positive reference (HPA tissue IHC) and titrate the catalog antibody within 1:50–1:200 (datasheet M03104).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RBM3 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 in most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03104) | |
| Positive control | Nasopharynx+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | Cold inducible (UniProt) | |
| Isoform / epitope | No annotated isoforms; chain spans residues 1–157 (UniProt) |
Section 1
Recommended RBM3 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet M03104) with published RBM3 protocols for prostate, breast, melanoma, and esophageal tissue (PMC3195697; PMC6158158; PMC3156749; PMC6233549).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M03104) |
| Fixation | Image fixative and duration unreported (datasheet M03104); 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 M03104); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03104) |
| Primary antibody | Rabbit monoclonal (clone 29R08) anti-RBM3, 1:50-1:200 (datasheet M03104) |
| Primary incubation | Overnight at 4 °C (datasheet M03104) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03104) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RBM3-positive staining in basal cells of nasopharynx (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet M03104). Use a paper’s retrieval conditions when following its protocol (PMC6158158; PMC6233549).
Section 2
What Is the Expected RBM3 Staining Pattern?
RBM3 is predominantly nuclear in tissue IHC (HPA: nuclear expression in most tissues), with nucleoplasmic localisation in ICC-IF (HPA: enhanced). Expect strong staining in nasopharyngeal basal cells and moderate staining in several epithelial and lymphoid cell types (HPA: High; Medium). UniProt also records cytoplasm and dendritic mRNA granules, so localisation can vary by context (UniProt P98179). RBM3 has no transmembrane segment (UniProt P98179 topology).
| Nuclear signal is strongest in nasopharyngeal basal cells, with weaker or patchier staining in other cells. | This fits the tissue reference: basal cells are High, while appendix glandular cells, cervical squamous cells and lymph-node germinal-center cells are Medium (HPA: tissue IHC). Judge intensity within identified cell types; HPA reports low tissue RNA specificity and only medium consistency between antibody staining and RNA expression (HPA: reliability Enhanced). |
| Staining is predominantly cytoplasmic, membranous or extracellular, with little nuclear signal. | This departs from the expected tissue pattern (HPA: nuclear expression in most tissues). Check morphology, counterstain, detection controls and whether the signal follows tissue edges or deposits (general IHC practice). Cytoplasmic RBM3 is biologically possible, so cytoplasmic signal alone is not proof of artefact (UniProt P98179: cytoplasm). Membrane localisation is unsupported by the supplied record (UniProt P98179 topology). |
| Strong staining appears in an HPA-listed undetected cell type, such as adipocytes or cardiomyocytes. | Treat this as a discordance to investigate, not an automatic biological negative: those specific cell types were Not detected in the cited tissue samples (HPA: adipose tissue; heart muscle). Review cell identification, antibody-specific staining and endogenous detection activity with appropriate controls (general IHC practice). HPA's Enhanced tissue reliability still allows sample-level variation (HPA: medium staining–RNA consistency). |
| A broad haze covers nuclei, cytoplasm and blank areas, obscuring cell boundaries. | Diffuse signal without a cell-type or nuclear pattern is difficult to score against HPA's nuclear reference (HPA: tissue IHC). Uneven reagent coverage, insufficient blocking or washing, excess detection reagent, or chromogen precipitate are possible workflow causes (general IHC practice). Compare a no-primary control and inspect unstained spaces before calling RBM3 positive (general IHC practice). |
| Nasopharyngeal basal-cell nuclei show no signal in a section expected to be positive. | The result conflicts with the High reference staining for that cell type (HPA: nasopharynx basal cells), but one absent result cannot identify the failed step. Confirm that basal cells are present, then check the antibody's IHC-P instructions, retrieval and detection controls (general IHC practice). The supplied sources do not establish RBM3-specific fixation sensitivity. |
💡Expected RBM3 appearanceCall a convincing IHC positive result when morphologically identified nasopharyngeal basal cells show prominent nuclear staining (HPA: High; nuclear expression); isolated membrane signal or staining confined to HPA-listed undetected cell types warrants control review before scoring (UniProt P98179 topology; HPA: tissue IHC).
| Tissue and cell-type reference | Use the named cell type, not the organ alone, for comparison: nasopharyngeal basal cells are High; lung type-I alveolar cells and lymph-node germinal-center cells are Medium; adipocytes are Not detected (HPA: tissue IHC). These are observed examples, not universal thresholds; HPA reports low tissue RNA specificity and medium staining–RNA consistency (HPA: reliability Enhanced). |
| Compartment and molecular topology | The primary tissue expectation is nuclear (HPA: tissue IHC), and ICC-IF places RBM3 in the nucleoplasm (HPA: enhanced subcellular location). UniProt also lists cytoplasm and dendritic mRNA granules, so assess unusual cytoplasmic staining with controls rather than rejecting it outright (UniProt P98179). No transmembrane segment or signal peptide supports a routine membrane or secreted pattern (UniProt P98179 topology; processing). |
| Antibody validation | HPA lists IHC as Enhanced for CAB062559 and Supported for HPA003624; ICC is Supported for both (HPA: antibody validation). These grades are antibody-specific and do not validate every staining condition or unexpected compartment. Interpret a discordant result alongside the chosen antibody's validation status and control sections (HPA: antibody validation; general IHC practice). |
| IF/ICC Q: What pattern is expected? | A: Nucleoplasmic staining is the reported main location; HPA has ICC-IF images in A-431, U2OS and BJ cells (HPA: subcellular). That observation helps interpret localisation but does not supply an IHC-P retrieval, dilution or detection protocol (HPA: subcellular). |
| Processing and modifications | UniProt lists one 1–157 chain, no signal peptide, no propeptide, no annotated glycosylation sites and no isoforms (UniProt P98179). It also records arginine methylation, but the supplied sources do not connect those modifications to a particular antibody epitope or staining intensity; avoid using them to explain a weak section (UniProt P98179). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear signal in nasopharyngeal basal cells (HPA: High). | The expected cells may be absent from the field, or an IHC step may have failed; the source data do not identify a specific failure (HPA: nasopharynx; general IHC practice). | Verify basal-cell morphology and section integrity, then review the catalog antibody's IHC-P instructions and run an appropriate positive control through the same retrieval and detection workflow (general IHC practice). |
| Signal is present but too weak to score in an expected-positive cell type (HPA: tissue IHC). | Weak detection may reflect section or workflow performance; HPA's Medium categories are observed levels, not guaranteed intensity in every specimen (HPA: tissue IHC; general IHC practice). | Compare a validated positive section in the same run and check antibody preparation, retrieval execution, incubation and chromogen development against the chosen IHC-P instructions (general IHC practice). Avoid assigning an RBM3-specific fixation effect from these sources. |
| Nuclei stain uniformly across cells expected to differ (HPA: High, Medium and Not detected examples). | Overdeveloped chromogen, excessive detection signal or background may obscure cell-type differences (general IHC practice); HPA's categories do not define a universal quantitative cutoff (HPA: tissue IHC). | Review development and wash steps, examine a no-primary control, and score only clearly identified cells with interpretable nuclear morphology (general IHC practice). Compare the same cell type across control and test sections. |
| Staining is concentrated at tissue edges or in empty spaces (general IHC practice). | Edge effects, drying or precipitate can produce spatial patterns unrelated to the nuclear tissue reference (general IHC practice; HPA: tissue IHC). | Inspect reagent coverage and chromogen preparation, repeat with even section wetting, and use a no-primary control to identify detection background (general IHC practice). Do not count acellular deposits as RBM3-positive cells. |
| Only cytoplasmic staining appears, without convincing nuclear staining (HPA: tissue IHC). | The result is discordant with HPA's predominant nuclear pattern, although UniProt permits cytoplasmic localisation (HPA: tissue IHC; UniProt P98179). Compartment misidentification or background is also possible (general IHC practice). | Check the nuclear counterstain and cell boundaries, compare a known-positive section, and assess no-primary staining (general IHC practice). Report a reproducible cytoplasmic pattern separately from the expected nuclear score. |
| A HPA-listed undetected cell type stains strongly (HPA: adipocytes, cardiomyocytes or bone-marrow hematopoietic cells). | The finding may reflect cell-identification error, antibody cross-reactivity, endogenous detection activity or sample variation; HPA provides an observed reference, not proof of absence in every specimen (HPA: tissue IHC; general IHC practice). | Recheck morphology, examine a no-primary control and compare the positive control in the same run (general IHC practice). If the signal persists, describe the cell type and compartment rather than calling the HPA reference disproven. |
Experiment evidence
IHC & IF images showing RBM3 staining
M03104 · IHCIHC analysis of RBM3 using anti-RBM3 antibody (M03104). RBM3 was detected in a paraffin-embedded section of human colorectal adenocarcinoma
M03104 · IHCIHC analysis of RBM3 using anti-RBM3 antibody (M03104). RBM3 was detected in a paraffin-embedded section of human testicular germ cell tumor
M03104 · IFIF analysis of RBM3 using anti-RBM3 antibody (M03104) and anti-Beta Tubulin antibody (M01857-3). RBM3 was detected in immunocytochemical sec
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000102317-RBM3/tissue/cerebral+cortex#imid_17884883
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000102317-RBM3/tissue/esophagus#imid_17884863
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000102317-RBM3/tissue/colon#imid_17884809
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000102317-RBM3/tissue/liver#imid_17884818
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003624; green = RBM3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003624; green = RBM3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RBM3 IHC & IF
🧪Run nasopharynx first; its basal cells should stain strongly (HPA: High in nasopharyngeal basal cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the positive slide, cells with little or no staining outside the basal compartment can serve as internal negative comparators, but their absence of RBM3 should not be presumed (HPA: High in basal cells).
Positive control tissue: Nasopharynx (Basal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RBM3 in A-431, U2OS, BJ [Human fibroblast], with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, plus matched RBM3-knockout material if available (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase and inspect background in the nasopharynx section before interpreting DAB staining (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected paraffin-section caption does not state a fixative (selected M03104 caption). Heat retrieval in EDTA at pH 8.0 is documented for the selected IHC example, but its necessity has not been established by the supplied evidence (selected M03104 caption). The evidence does not establish that frozen sections or IF are easier; for IF, nucleoplasmic signal is a localization reference, while background in nasopharyngeal tissue should be checked on the actual section (HPA: Nucleoplasm, enhanced; standard IF practice).
HPA tissue IHC evidence for RBM3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Nasopharynx | Basal cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RBM3 IHC Tips
Troubleshoot RBM3 staining in paraffin sections by checking retrieval, nuclear localisation, controls and scoring before interpreting chromogenic signal.
How should I optimise retrieval when RBM3 nuclear staining is weak?
Start with heat-mediated antigen retrieval in EDTA pH 8.0 for paraffin sections (datasheet M03104). The selected tissue image used this retrieval before overnight incubation with the catalog antibody at 1:50 and 4°C (datasheet M03104). If nuclei remain weak, compare a controlled range of heating times on adjacent sections while keeping antibody concentration and detection constant (standard IHC practice). Include a known positive section, such as nasopharyngeal basal cells, and inspect tissue integrity alongside staining intensity (HPA: High in nasopharyngeal basal cells; standard IHC practice). Score nuclear signal separately from diffuse cytoplasmic colour because RBM3 is predominantly nuclear in tissue IHC (HPA: nuclear expression in most tissues).
Could fixation explain variable RBM3 staining between paraffin blocks?
Target-specific fixation sensitivity for RBM3 is unknown: the selected paraffin-section caption does not report a fixative (datasheet M03104). Record each block’s fixative and fixation duration, then compare sections processed with the same retrieval and staining run (standard IHC practice). Include matched positive tissue in each run so variation in chromogenic development does not masquerade as a fixation effect (standard IHC practice). Examine morphology and nuclear counterstain before adjusting retrieval, because damaged or poorly preserved nuclei complicate RBM3 assessment (standard IHC practice; HPA: nuclear expression in most tissues). Do not infer fixation tolerance from RBM3 localisation or methylation annotations (UniProt P98179: localisation and modified residues).
Should I score cytoplasmic RBM3 staining as positive in paraffin sections?
Prioritise nuclear staining: tissue IHC shows nuclear expression in most tissues, and subcellular imaging places RBM3 in the nucleoplasm (HPA: tissue IHC profile; HPA: enhanced nucleoplasmic localisation). Cytoplasmic RBM3 is biologically plausible, with UniProt also annotating cytoplasm and dendritic mRNA granules (UniProt P98179: subcellular locations). Nevertheless, record nuclear and cytoplasmic staining separately rather than merging them into one positive score (standard IHC practice). Compare any predominantly cytoplasmic pattern with adjacent tissue, a no-primary control and the expected nuclear pattern before interpreting it (standard IHC practice; HPA: tissue IHC profile). RBM3 has no annotated transmembrane segment, so crisp membrane staining warrants particular scrutiny (UniProt P98179: topology).
How can epitope uncertainty affect RBM3 IHC interpretation?
The supplied record lists one RBM3 chain of 157 amino acids and no annotated isoforms, but does not map the catalog antibody’s epitope (UniProt P98179: processing and isoforms; datasheet M03104). Its RNA recognition motif spans residues 6–84, and several arginine methylation annotations occur elsewhere or near that region (UniProt P98179: domain and modified residues). Without an epitope map, neither differential recognition of a modified residue nor its exclusion can be established (UniProt P98179: modified residues; datasheet M03104). Keep retrieval and antibody conditions consistent across specimens, and confirm unexpected compartment patterns with an independently validated reagent if available (standard IHC practice). Treat staining differences as assay observations until specificity has been tested (standard IHC practice).
How should I investigate RBM3 by IF alongside a cell-type marker?
For a separate IF assay, pair RBM3 with a marker that identifies the cell population being assessed and use distinct detection channels (standard IF practice). Choose fluorophores after checking tissue autofluorescence in unstained sections, reserving the cleaner channel for weaker signal (standard IF practice). RBM3 is predominantly nucleoplasmic, so use fixation-compatible permeabilisation that permits antibody access to intracellular epitopes and assess colocalisation against a nuclear counterstain (HPA: enhanced nucleoplasmic localisation; standard IF practice). Include single-stain and no-primary controls to check channel bleed-through and background (standard IF practice). The supplied paraffin-section caption establishes chromogenic IHC conditions, while IF performance of catalog antibody M03104 is not established by that caption (datasheet M03104).
What should I check when RBM3 DAB staining looks diffuse?
First compare a no-primary section with the stained section to identify signal arising from the detection system or tissue (standard IHC practice). Block endogenous peroxidase before HRP detection, then check that DAB development is stopped before background obscures nuclear detail (standard IHC practice). The selected paraffin-section image used 10% goat serum blocking, overnight antibody incubation at 1:50 and 4°C, and HRP with DAB (datasheet M03104). If background persists, optimise blocking, washing and antibody concentration on matched sections while retaining a positive control (standard IHC practice). Interpret diffuse colour cautiously because the expected tissue pattern is chiefly nuclear (HPA: nuclear expression in most tissues).
How should I quantify RBM3 across tissue sections with mixed staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; RBM3 tissue IHC is chiefly nuclear (HPA: nuclear expression in most tissues). For nuclear signal, report the percentage of positive cells or an H-score from 0–300, using fixed intensity thresholds across samples (standard IHC scoring practice). If counting spatially distinct cells, report positive cells per mm² of viable tissue and document excluded areas (standard IHC quantification practice). Normalise counts to the relevant cell population or viable tissue area, and compare only sections processed and imaged under matched conditions (standard IHC practice). Keep cytoplasmic scores separate if that signal is being investigated (UniProt P98179: cytoplasmic localisation; standard IHC practice).
How can I distinguish true RBM3 signal from staining artefact?
A credible result should include assessable nuclear staining in the expected cell population and agreement with a processed positive control (HPA: nuclear expression in most tissues; standard IHC practice). Nasopharyngeal basal cells show high staining, whereas adipocytes were not detected in the supplied tissue profile; these are reference patterns, not absolute thresholds (HPA: nasopharynx and adipose tissue). Inspect edge staining, necrotic areas and no-primary controls before calling isolated DAB deposits positive (standard IHC practice). Unexpected membrane-only staining deserves scrutiny because RBM3 has no transmembrane segment (UniProt P98179: topology). Endogenous peroxidase can contribute chromogenic signal, so confirm adequate blocking and compare the relevant control section (standard IHC practice).
Boster reagents
Best RBM3 / RNA-binding protein 3 IHC Antibodies
Anti-RBM3 M03104 has IHC images from paraffin-embedded human tumor sections (M03104 IHC captions) and IF data from A549 cells (M03104 IF caption); human and mouse reactivity is listed (catalog: M03104).
Real IHC data
Anti-RBM3 Rabbit Monoclonal Antibody
Cat # M03104
M03104 is listed for IHC and IF/ICC (catalog: M03104 applications), with IHC images from paraffin-embedded human colorectal adenocarcinoma, testicular germ cell tumor, liver cancer, and lung adenocarcinoma sections (M03104 IHC captions). Its IF image shows A549 cells (M03104 IF caption), and the catalog lists human and mouse reactivity (catalog: M03104 reactivity).
Which to pick: Choose M03104 for paraffin-section IHC: it is a rabbit monoclonal listed for IHC (catalog: M03104), and its own captions document EDTA retrieval at pH 8.0 and 1:50 primary dilution (M03104 IHC captions). The IHC captions do not report the fixative (M03104 IHC captions). Choose M03104 for IF/ICC based on its application listing and A549 IF image (catalog: M03104 applications; M03104 IF caption); for cross-species work, it lists human and mouse reactivity, while the supplied IHC images show human tissue (catalog: M03104 reactivity; M03104 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P98179 (RBM3_HUMAN, RNA-binding protein 3).
- Human Protein Atlas. RBM3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RBM3 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RBM3 antibody validation summary (2 antibodies).
- High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression. Diagnostic pathology 2011 — PMC3195697.
- High RNA-binding Motif Protein 3 Expression Is Associated with Improved Clinical Outcomes in Invasive Breast Cancer. Journal of breast cancer 2018 — PMC6158158.
- Low RBM3 protein expression correlates with tumour progression and poor prognosis in malignant melanoma: an analysis of 215 cases from the Malmö Diet and Cancer Study. Journal of translational medicine 2011 — PMC3156749.
- Reduced RBM3 expression is associated with aggressive tumor features in esophageal cancer but not significantly linked to patient outcome. BMC cancer 2018 — PMC6233549.
- PubMed PMID:8634703 — UniProt-cited evidence.
- PubMed PMID:15772651 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RBM3 (RNA-binding protein 3). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03104). Validated IHC antibody: M03104 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Nasopharynx (Basal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M03104); 2. Fixation: Image fixative and duration unreported (datasheet M03104); 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 M03104); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03104); 8. Primary antibody: Rabbit monoclonal (clone 29R08) anti-RBM3, 1:50-1:200 (datasheet M03104); 9. Primary incubation: Overnight at 4 °C (datasheet M03104). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; chain spans residues 1–157 (UniProt); Regulation: Cold inducible (UniProt). Evidence: UniProt P98179, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet M03104, PMC3195697, PMC6158158. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RBM3/immunohistochemistry-ihc-guide.