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RRM2B / Ribonucleoside-diphosphate reductase subunit M2 B · IHC design guide
Design Immunohistochemistry for RRM2B
Plan RRM2B paraffin-section IHC around cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Use colon glandular cells as a high-staining reference and heart cardiomyocytes as a tissue reference where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RRM2B 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 | Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cells in most tissues show cytoplasmic and nuclear signal (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Heart muscle+2 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 | DNA damage may shift signal toward nuclei (UniProt) | |
| Regulation | Responds to DNA damage through TP53 (UniProt) | |
| Isoform / epitope | 6 isoforms; assess epitope coverage across variants (UniProt) |
Section 1
Recommended RRM2B IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published RRM2B IHC workflows (PMC3085570; PMC12852120; PMC3492868; PMC6788385).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03055-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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-RRM2B, 1 μg/mL (datasheet A03055-1) |
| 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 | RRM2B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare the published citrate or steam retrieval conditions if needed (PMC12852120; PMC3492868).
Section 2
What Is the Expected RRM2B Staining Pattern?
RRM2B should appear in the cytoplasm and nuclei of appropriate cells, with staining interpreted by cell type and compartment rather than by tissue alone (HPA tissue IHC: cytoplasmic and nuclear expression in most tissues; UniProt Q7LG56: cytoplasm, nucleus, no transmembrane segment). HPA rates the tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC: Approved).
| Cytoplasmic and nuclear staining in colon glandular cells or bronchial respiratory epithelial cells. | This fits reported high staining in those cells (HPA tissue IHC: High in colon glandular and bronchial respiratory epithelial cells). Assess the intended cells separately from adjacent tissue. |
| A sharp membrane rim or extracellular deposit dominates the slide. | That is outside the reported cytoplasmic and nuclear pattern (HPA tissue IHC; UniProt Q7LG56: no transmembrane segment). Review morphology, detection background and antibody specificity before calling it RRM2B. |
| Strong staining appears in cardiomyocytes, smooth muscle cells or vaginal squamous epithelial cells. | These cells are reported as not detected (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; compare a no-primary control before assigning the signal to RRM2B (standard IHC practice). |
| Brown signal spreads evenly across cells and surrounding tissue, obscuring boundaries. | A diffuse field cannot establish the reported cellular pattern (HPA tissue IHC: cytoplasmic and nuclear expression). Check background controls and distinguish deposits from cell-associated stain (standard IHC practice). |
| Colon glandular cells or bone marrow hematopoietic cells show no discernible signal. | Both are reported high-staining examples (HPA tissue IHC). First confirm tissue identity and an interpretable positive control, then review section processing and the catalog antibody’s IHC-P directions (standard IHC practice). |
💡Expected RRM2B appearanceCall a result positive when the relevant cells show discernible cytoplasmic and nuclear staining, especially in reported high-staining colon glands or bone marrow hematopoietic cells (HPA tissue IHC); a membrane rim or diffuse extracellular deposit alone does not match the reported location (HPA tissue IHC; UniProt Q7LG56 topology).
| Tissue and cell selection | Use reported high-staining cells as positive references and reported not-detected cells as contrasts (HPA tissue IHC). Score named cell populations: a tissue-level label does not make every cell positive. |
| Subcellular distribution | Cytoplasm and nucleus are both reported (UniProt Q7LG56; HPA tissue IHC). UniProt reports movement from cytoplasm to nucleus after DNA damage; a single section cannot establish that movement without matched experimental conditions. |
| HPA evidence level | The tissue profile is Approved with medium RNA–staining consistency (HPA tissue IHC). HPA028812 and CAB006854 each have IHC Approved status (HPA antibodies); this record does not assign either an IHC Enhanced designation. |
| Skeletal muscle interpretation | UniProt describes high skeletal-muscle expression, while HPA reports low staining in myocytes (UniProt Q7LG56 tissue specificity; HPA tissue IHC: Low in skeletal-muscle myocytes). Do not use skeletal muscle as an unequivocal high-staining IHC reference. |
| Protein forms and assay scope | UniProt lists six isoforms, a single annotated chain spanning residues 1–351, no signal peptide and no glycosylation sites (UniProt Q7LG56). Epitope coverage of individual isoforms is not supplied; do not infer it from staining. |
| Detection background | Endogenous enzyme activity can contribute chromogenic signal, and inadequate blocking or washing can raise background (standard IHC practice). These are general assay possibilities, not demonstrated RRM2B-specific effects. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining cells remain blank. | The run may have failed, or the chosen cells may be absent or poorly represented; the stated reference is cell-specific (HPA tissue IHC). | Check tissue identity and morphology, review a positive control, then follow the catalog antibody’s IHC-P directions for retrieval, dilution and detection (standard IHC practice). No target-specific retrieval condition is supplied here. |
| Only a membrane outline is prominent. | Membrane-restricted staining conflicts with the reported intracellular distribution (HPA tissue IHC; UniProt Q7LG56 topology). | Inspect a no-primary control and staining morphology; repeat with appropriate controls if the rim persists (standard IHC practice). Do not score a rim alone as positive. |
| Reported not-detected cell types stain strongly. | Cross-reactivity or endogenous chromogenic activity is possible; HPA reports no detection in cardiomyocytes, smooth muscle cells and vaginal squamous epithelial cells (HPA tissue IHC). | Compare no-primary and positive controls, then reassess cell identification and the catalog antibody’s IHC-P detection steps (standard IHC practice). |
| The whole section has diffuse brown haze. | General detection background may obscure cell boundaries (standard IHC practice). Haze alone does not demonstrate the reported cytoplasmic and nuclear pattern (HPA tissue IHC). | Review no-primary control, blocking, washes and counterstain; score only cell-associated signal with readable morphology (standard IHC practice). |
| Skeletal-muscle myocytes stain weakly despite a high-expression annotation. | The sources differ in measurement: UniProt reports high skeletal-muscle expression, while HPA tissue IHC reports low staining in myocytes (UniProt Q7LG56; HPA tissue IHC). | Document the observed IHC level and use a reported high-staining cell population for the positive reference (HPA tissue IHC). Do not reinterpret weak myocyte staining as an assay failure by itself. |
| What localisation should an IF/ICC check show? | HPA’s ICC-IF summary places RRM2B mainly in the nucleoplasm, with additional cytosolic signal (HPA subcellular: enhanced). Both listed antibodies have ICC Supported status (HPA antibodies). | Compare nuclear and cytosolic signal with suitable IF controls (standard IF practice). Use the separate IF/ICC guide for its workflow; the available IHC-P directions are not an IF protocol. |
Experiment evidence
IHC & IF images showing RRM2B staining
A03055-1 · IHCImmunohistochemistry of p53R2 in human lung tissue with p53R2 antibody at 1 μg/mL.
A03055-1 · IFImmunofluorescence of p53R2 in Human Lung tissue with p53R2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000048392-RRM2B/tissue/cerebral+cortex#imid_1876836
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000048392-RRM2B/tissue/colon#imid_1876778
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000048392-RRM2B/tissue/liver#imid_1876771
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000048392-RRM2B/tissue/kidney#imid_1876780
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028812; green = RRM2B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028812; green = RRM2B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RRM2B IHC & IF
🧪Run bone marrow first and require staining in hematopoietic cells (HPA: High in bone marrow hematopoietic cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); on the positive slide, count neighboring unstained cells as internal background comparators only after confirming their lack of specific staining, since HPA does not identify a negative bone marrow cell type (HPA: bone marrow row).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RRM2B in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and irrelevant-primary controls matched to the primary antibody’s host species, isotype and monoclonal or polyclonal format (standard IHC practice). Confirm specificity with RRM2B knockout material or a validated immunizing-peptide block (standard IHC practice). For chromogenic bone marrow IHC, quench endogenous peroxidase because myeloid cells can produce background signal; assess endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or retrieval requirement is reported in the supplied evidence; the selected A03055-1 lung IHC caption gives 1 µg/mL but leaves the fixative unreported (selected-SKU tissue-IHC caption). HPA provides IF/ICC images and reports mainly nucleoplasmic, with additional cytosolic, localization, but the evidence does not establish whether IF or frozen sections are easier than paraffin IHC (HPA: subcellular). Endogenous peroxidase in bone marrow myeloid cells can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for RRM2B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
Section 3
Advanced RRM2B IHC Tips
Troubleshoot RRM2B staining in paraffin sections by checking retrieval, compartment, cell type, controls and scoring before interpreting chromogenic signal.
How should I adjust retrieval when RRM2B staining is weak or diffuse?
Use Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min as this page’s starting condition (page retrieval rule: nuclear antigen). If staining is weak, compare an adjacent section with a shorter retrieval time before changing buffer, because excessive heating can damage morphology and increase diffuse staining (standard IHC practice). Keep section thickness, cooling, antibody concentration and chromogen development constant across the comparison so that retrieval is the variable being assessed (standard IHC practice). Evaluate both cytoplasmic and nuclear staining in the same cell type; either compartment can be plausible for RRM2B (UniProt Q7LG56 localisation; HPA tissue IHC profile).
Could fixation be masking RRM2B in my paraffin sections?
Target-specific fixation sensitivity is unknown from the supplied evidence; the selected antibody’s lung tissue caption reports 1 µg/mL but does not state a fixative (A03055-1 tissue-IHC caption). Record the actual fixative and fixation interval for each specimen before attributing weak staining to fixation (standard IHC practice). Compare sections processed with the same retrieval, antibody concentration and detection timing, and include a tissue control in each run to identify a broader processing failure (standard IHC practice). Avoid treating a difference between specimens as a fixation effect until specimen handling and detection variables have been checked (standard IHC practice).
Should RRM2B appear in nuclei, cytoplasm, or both?
Score nuclear and cytoplasmic staining separately: RRM2B is reported in both compartments, with movement from cytoplasm to nucleus after DNA damage (UniProt Q7LG56 localisation). Tissue IHC shows cytoplasmic and nuclear expression in most tissues, while cellular imaging reports mainly nucleoplasmic signal with additional cytosolic signal (HPA tissue IHC profile; HPA subcellular summary). Check that nuclei are intact and counterstained before calling a weak nuclear chromogen deposit positive, and compare compartment patterns within the same cell type (standard IHC practice). A change in nuclear staining alone does not establish DNA damage; interpret it alongside matched controls and independent evidence of the experimental condition (UniProt Q7LG56 localisation; standard IHC practice).
Can one IHC stain distinguish RRM2B isoforms or reveal epitope masking?
RRM2B has 6 listed isoforms, but the supplied antibody caption does not identify its epitope or establish isoform-specific staining (UniProt Q7LG56 isoforms; A03055-1 tissue-IHC caption). Treat chromogenic signal as antibody-reactive RRM2B unless the antibody’s epitope and its presence in each isoform are independently established (standard IHC interpretation). The protein has no annotated transmembrane segment, signal peptide, glycosylation sites or modified residues, so these annotations do not identify an epitope to target during retrieval (UniProt Q7LG56 topology and processing). If an expected compartment stains poorly, compare controlled retrieval conditions and an independent antibody with a documented epitope before assigning the discrepancy to an isoform (standard IHC practice).
How can I assess RRM2B localisation by multiplex IF alongside this IHC result?
For the separate IF/ICC workflow, pair RRM2B with a marker for the cell type being evaluated and a nuclear counterstain so compartment and cell identity can be assessed together (standard IF practice). Choose a fluorophore channel after inspecting tissue autofluorescence and use single-stain controls to set exposure and assess channel bleed-through (standard IF practice). Permeabilise sufficiently for access to intracellular nuclear and cytosolic epitopes; the supplied record reports no transmembrane segment and places RRM2B in both compartments (UniProt Q7LG56 topology and localisation; standard IF practice). Optimise IF fixation and permeabilisation directly, because the selected tissue-IHC caption does not provide an IF/ICC fixation condition (A03055-1 tissue-IHC caption; standard IF practice).
What controls help separate RRM2B staining from chromogenic background?
Run a no-primary section and inspect its chromogen deposits before interpreting faint RRM2B signal; it helps reveal detection-system or endogenous-enzyme background (standard IHC practice). For peroxidase-based detection, include a peroxidase block and keep DAB development consistent across test and control sections (standard IHC practice). Compare suspect staining with a matched tissue region showing interpretable cell morphology, rather than scoring deposits in folds, edges or damaged areas (standard IHC practice). Nuclear and cytoplasmic staining are both biologically plausible, so compartment alone cannot certify specificity; require a clean control and a reproducible cell-associated pattern (UniProt Q7LG56 localisation; HPA tissue IHC profile; standard IHC practice).
How should I quantify RRM2B across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because RRM2B can appear in both nucleus and cytoplasm (UniProt Q7LG56 localisation; HPA tissue IHC profile). For each compartment, record the percentage of positive cells and intensity categories, then calculate an H-score using percentages at 0–3 intensity if that scale fits the study (standard IHC practice). Normalise the positive count to the number of evaluable cells of that type, or report positive-cell density per mm² of evaluable tissue when area is the denominator (standard IHC practice). Apply identical thresholds, exclusion rules and image-acquisition settings to every section, and report nuclear and cytoplasmic results separately (standard IHC practice).
When is an apparent RRM2B-positive area likely to be artefact?
Look for cell-associated nuclear or cytoplasmic signal with preserved morphology; both compartments are reported for RRM2B (UniProt Q7LG56 localisation; HPA tissue IHC profile). Treat signal in a compartment outside that pattern, extracellular deposits, or staining concentrated at section edges and necrotic areas as suspect until controls resolve it (standard IHC practice). Check which cells actually stain: high glandular-cell staining is reported in colon, whereas smooth muscle cells were not detected in the cited tissue survey (HPA tissue IHC: colon glandular cells High; smooth muscle cells Not detected). If a no-primary control retains chromogen, investigate endogenous enzyme activity or detection background before calling the sample positive (standard IHC practice).
Boster reagents
Best RRM2B / Ribonucleoside-diphosphate reductase subunit M2 B IHC Antibodies
A03055-1 has IHC and IF images from human lung tissue (catalog image captions). Human, mouse and rat reactivity is listed (catalog: reactivity); the images document human tissue only (catalog image captions).
Real IHC data
Anti-p53R2 RRM2B Antibody
Cat # A03055-1
A03055-1 is listed for IHC-P and IF (catalog: applications). Its IHC and IF images show human lung tissue at 1 μg/mL and 20 μg/mL, respectively (catalog image captions).
Which to pick: Choose A03055-1 for paraffin-section IHC (catalog: IHC-P); its own IHC caption documents human lung tissue at 1 μg/mL, while the fixative is unreported (catalog: IHC image caption). For tissue IF, choose A03055-1 based on its human lung IF image at 20 μg/mL; ICC-specific validation is unreported (catalog: IF image caption and applications). For mouse or rat samples, A03055-1 lists reactivity with both species, but its IHC and IF images show human tissue only (catalog: reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q7LG56 (RIR2B_HUMAN, Ribonucleoside-diphosphate reductase subunit M2 B).
- Human Protein Atlas. RRM2B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RRM2B subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. RRM2B antibody validation summary (2 antibodies).
- Ribonucleotide reductase small subunit M2B prognoses better survival in colorectal cancer. Cancer research 2011 — PMC3085570.
- Integrative clinico-molecular analysis reveals actionable subtypes and biomarkers in lung adenocarcinoma. Cell discovery 2026 — PMC12852120.
- RRM2B suppresses activation of the oxidative stress pathway and is up-regulated by p53 during senescence. Scientific reports 2012 — PMC3492868.
- Integrative Analysis Reveals Across-Cancer Expression Patterns and Clinical Relevance of Ribonucleotide Reductase in Human Cancers. Frontiers in oncology 2019 — PMC6788385.
- PubMed PMID:10716435 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RRM2B (Ribonucleoside-diphosphate reductase subunit M2 B). Expected localisation Cytoplasmic and nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A03055-1 at 1 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Heart muscle. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A03055-1); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 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-RRM2B, 1 μg/mL (datasheet A03055-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: DNA damage may shift signal toward nuclei (UniProt); Isoform / epitope: 6 isoforms; assess epitope coverage across variants (UniProt); Regulation: Responds to DNA damage through TP53 (UniProt). Evidence: UniProt Q7LG56, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03055-1, PMC3085570, PMC12852120. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RRM2B/immunohistochemistry-ihc-guide.