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RCC1 / Regulator of chromosome condensation · IHC design guide
Design Immunohistochemistry for RCC1
Plan RCC1 staining in paraffin sections around the general nuclear tissue pattern (HPA tissue IHC). Use the catalog antibody’s documented IHC-P conditions and account for RCC1 binding to mitotic chromosomes when interpreting staining (datasheet A02719-1; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RCC1 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 | General nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across many tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02719-1) | |
| 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 | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Mitotic chromosome binding may change the nuclear pattern (UniProt) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | Two isoforms; epitope coverage unspecified (UniProt) |
Section 1
Recommended RCC1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A02719-1) with four published RCC1 paraffin-section methods (PMC5964113; PMC8051708; PMC8674189; PMC12804135).
| Sample | Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A02719-1) |
| Fixation | Image fixative and duration unreported (datasheet A02719-1); 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 A02719-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02719-1) |
| Primary antibody | Rabbit anti-RCC1, 0.5-1μg/ml (datasheet A02719-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02719-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02719-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RCC1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A02719-1); assess published citrate methods separately (PMC8051708; PMC8674189).
Section 2
What Is the Expected RCC1 Staining Pattern?
RCC1 should stain predominantly in interphase nuclei and associate with mitotic chromosomes (UniProt P18754 localization). In paraffin-section IHC, expect nuclear staining across many cell types, with high staining reported in several glandular and epithelial populations (HPA: general nuclear expression; High in listed glandular and epithelial cells). HPA rates the tissue pattern Enhanced, with high consistency between staining and RNA expression (HPA: Enhanced). RCC1 has no transmembrane segment (UniProt P18754 topology).
| Nuclear staining in breast or adrenal glandular cells. | This matches reported high staining in these cells and RCC1’s predominant interphase location (HPA: High; UniProt P18754 localization). Judge the stained nuclei against the tissue’s counterstain and local background (general IHC practice). |
| Staining follows condensed chromosomes in a mitotic cell. | Chromosome-associated RCC1 is expected during mitosis (UniProt P18754 localization). Interpret this alongside surrounding interphase nuclei: a small mitotic focus can look different from the broader nuclear pattern (UniProt P18754 localization; general IHC practice). |
| Strong cytoplasmic staining dominates while nuclei appear unstained. | Cytoplasm is listed as a possible location, so cytoplasmic signal alone is not proof of artefact (UniProt P18754 localization). A cytoplasm-only result conflicts with the predominant interphase nuclear pattern; check morphology and controls before scoring it as RCC1 (UniProt P18754 localization; general IHC practice). |
| Strong signal is confined to an unexpected cell population while reference cells are blank. | For example, high cardiomyocyte staining with absent breast glandular staining reverses the reported contrast (HPA: Low in cardiomyocytes; High in breast glandular cells). Consider cross-reactivity or endogenous detection activity, and verify with detection controls (general IHC practice). |
| No discernible nuclear signal in a high-staining reference tissue. | Absent signal in breast glandular cells or bronchial respiratory epithelial cells disagrees with the reported high pattern (HPA: High in both). Confirm tissue preservation and controls, then review retrieval and detection conditions as general IHC checks; the sources do not report RCC1-specific fixation sensitivity. |
💡Expected RCC1 appearanceCall a positive result when glandular or epithelial cells show clear, predominantly nuclear chromogenic staining, with high signal possible in HPA-listed cells (HPA: High; UniProt P18754 localization); widespread diffuse background or nuclear-negative, cytoplasm-dominant staining is suspect (general IHC practice; UniProt P18754 localization).
| Cell-cycle location | RCC1 is predominantly nuclear in interphase and binds mitotic chromosomes (UniProt P18754 localization). Compare each staining pattern with nuclear morphology before treating variation within a section as technical failure (general IHC practice). |
| Tissue and cell population | HPA reports general nuclear expression and low RNA tissue specificity, yet its sampled cell populations range from High in breast glandular cells to Low in cardiomyocytes (HPA: tissue IHC). A low-staining cell population is a weak negative control, not evidence that the tissue lacks RCC1. |
| Antibody validation | HPA027573 and CAB015413 have Enhanced IHC validation; HPA027574 is Supported for IHC (HPA: antibody validation). These ratings support the reported pattern for those antibodies; they do not establish that every catalog antibody will reproduce it. |
| Molecular features and epitope uncertainty | UniProt lists two isoforms, no signal peptide or transmembrane segment, and several N-terminal modifications (UniProt P18754). The supplied records do not map the catalog antibody’s epitope, so they cannot predict isoform recognition, retrieval needs, or fixation sensitivity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Reference tissue shows no nuclear signal. | The result conflicts with reported high staining in breast glandular cells (HPA: High); the cause cannot be assigned from HPA alone. | Check a known-positive section and detection controls, then review the antibody’s stated IHC conditions and the run’s retrieval and detection steps (general IHC practice). Do not infer an RCC1-specific fixation effect. |
| Brown signal spreads across nuclei, cytoplasm and empty areas. | Diffuse staining without cellular boundaries suggests nonspecific background or detection activity (general IHC practice). | Compare with a section omitting primary antibody; review blocking, washes and chromogen development (general IHC practice). Score RCC1 only where the cellular pattern is interpretable. |
| Cytoplasmic signal is stronger than nuclear signal throughout the section. | A cytoplasm-only pattern differs from predominant interphase nuclear localization, although cytoplasm is also listed (UniProt P18754 localization). | Use the counterstain to confirm compartment boundaries and compare a high-staining reference population plus detection controls (general IHC practice; HPA: High in listed cells). |
| Low-staining cells look stronger than the expected reference cells. | This reverses an HPA contrast if, for example, cardiomyocytes exceed breast glandular cells (HPA: Low in cardiomyocytes; High in breast glandular cells). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Inspect tissue identity and morphology, compare a no-primary control, and avoid calling the unexpected signal specific without corroboration (general IHC practice). |
| Only occasional condensed structures stain strongly. | RCC1 binds mitotic chromosomes, which can produce a concentrated pattern in mitotic cells (UniProt P18754 localization). | Check whether the structures are mitotic chromosomes and assess interphase nuclei separately (UniProt P18754 localization; general IHC practice). |
| Q: Should an IF/ICC image show the same chromogenic pattern? | HPA reports nucleoplasmic localization by ICC-IF, while this guide’s primary application is paraffin-section IHC (HPA: nucleoplasm, enhanced; HPA: tissue IHC). | A: Expect nucleoplasmic fluorescence in ICC-IF (HPA: nucleoplasm, enhanced). Use the separate IF/ICC guide for that application; this section supplies no IF/ICC protocol. |
Experiment evidence
IHC & IF images showing RCC1 staining
A02719-1 · IHCIHC analysis of RCC1 using anti-RCC1 antibody (A02719-1). RCC1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat
A02719-1 · IHCIHC analysis of RCC1 using anti-RCC1 antibody (A02719-1). RCC1 was detected in paraffin-embedded section of human rectal cancer tissue. Heat
A02719-1 · IFIF analysis of RCC1 using anti-RCC1 antibody (A02719-1). RCC1 was detected in immunocytochemical section of Hela cells. Enzyme antigen retri
M02719-2 · IHCIHC analysis of RCC1 using anti-RCC1 antibody (M02719-2). RCC1 was detected in a paraffin-embedded section of human thyroiditis tissue. Heat
M02719-2 · IHCIHC analysis of RCC1 using anti-RCC1 antibody (M02719-2). RCC1 was detected in a paraffin-embedded section of human colorectal adenocarcinom
M02719-2 · IFIF analysis of RCC1 using anti-RCC1 antibody (M02719-2). RCC1 was detected in an immunocytochemical section of A431 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000180198-RCC1/tissue/cerebral+cortex#imid_7162472
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000180198-RCC1/tissue/colon#imid_7162390
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000180198-RCC1/tissue/liver#imid_7162399
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000180198-RCC1/tissue/kidney#imid_7162408
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB015413; green = RCC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB015413; green = RCC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RCC1 IHC & IF
🧪Run adrenal gland first; glandular cells should stain for RCC1 (HPA: High in adrenal gland glandular cells), with predominantly nuclear signal in interphase cells (UniProt P18754 subcellular location). HPA detects RCC1 in all 45 scored tissues and lists no negative tissue (HPA: no negative rows); use no-primary and isotype controls, and treat cells on the positive slide with background-only staining as internal background references, not validated RCC1-negative cells.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: RCC1 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 RCC1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, rabbit IgG matched to the primary antibody’s format and concentration, and RCC1-knockout material or immunizing-peptide competition if available (caption: rabbit primary antibody). For adrenal sections, check endogenous peroxidase and biotin background before SABC/DAB detection (caption: SABC/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A02719-1 paraffin-section caption does not state a fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is a documented starting condition, but its necessity is unestablished (caption: heat-mediated EDTA retrieval). Paraffin IHC has a documented workflow (caption: paraffin section); the supplied evidence does not establish whether frozen sections or IF are easier or identify an adrenal-specific artefact (HPA: adrenal gland High; HPA: nucleoplasm enhanced in ICC-IF).
HPA tissue IHC evidence for RCC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RCC1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RCC1 IHC Tips
Troubleshoot RCC1 staining in paraffin sections by checking retrieval, nuclear localisation, controls, and scoring before interpreting signal (datasheet A02719-1; HPA tissue IHC).
What retrieval should I try first when RCC1 nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02719-1). The selected tissue image used this retrieval before 1 μg/ml primary antibody overnight at 4°C, so keep those conditions aligned when troubleshooting (datasheet A02719-1). Check whether nuclei in viable tissue gain crisp DAB signal while a no-primary section stays clear (standard IHC practice; HPA: general nuclear expression). If signal remains weak, vary heating duration in small steps and compare adjacent sections processed together, recording tissue damage as well as staining (standard IHC practice).
Could fixation explain inconsistent RCC1 staining across paraffin blocks?
Target-specific fixation sensitivity for RCC1 is unknown from the supplied evidence; the selected paraffin-section caption does not state its fixative (datasheet A02719-1). Record each block's fixative and fixation duration, then compare sections with matched thickness, retrieval, and detection conditions (standard IHC practice). A controlled trial using routinely fixed material can show whether processing correlates with weak nuclear staining, but cannot establish an RCC1-specific fixation mechanism by itself (standard IHC practice). Include a consistently staining tissue section in each run and assess nuclear morphology before changing primary-antibody concentration (HPA: general nuclear expression; standard IHC practice).
How should RCC1 staining appear in interphase and mitotic cells?
Expect predominantly nuclear staining in interphase cells and staining associated with mitotic chromosomes (UniProt P18754: localisation). HPA describes a general nuclear tissue pattern and an enhanced nucleoplasmic IF location, supporting nuclear signal as the main readout in paraffin-section IHC (HPA: tissue IHC and subcellular). Evaluate DAB against a nuclear counterstain, keeping mitotic figures distinct from interphase nuclei when reviewing the section (standard IHC practice). RCC1 also has a cytoplasmic annotation, so investigate convincing cytoplasmic staining with controls and morphology instead of treating every extranuclear pixel as a positive cell (UniProt P18754: localisation; standard IHC practice).
Could RCC1 isoforms or modified residues affect staining with this antibody?
RCC1 has 2 annotated isoforms, but the supplied caption does not map the catalog antibody's epitope or establish isoform selectivity (UniProt P18754: isoforms; datasheet A02719-1). The record lists alternative methylation or phosphorylation at residue 2 and phosphorylation at residue 11; their effects on this antibody's IHC staining are unknown (UniProt P18754: modified residues). Do not assign a missing nuclear signal to one isoform or modification without epitope information and an independent validation experiment (standard IHC practice). First confirm comparable retrieval, tissue integrity, and nuclear staining in a run control (datasheet A02719-1; HPA: general nuclear expression; standard IHC practice).
How can IF help assess an ambiguous RCC1 IHC pattern?
Use IF as a separate localisation check: RCC1 is mainly nucleoplasmic in the supplied IF record, while paraffin IHC shows general nuclear expression (HPA: subcellular; HPA: tissue IHC). Multiplex with a marker for the expected cell type in the specimen and a nuclear counterstain, then assess RCC1 within those cells rather than across the whole image (standard IF practice). Choose a fluorophore in a channel with low measured tissue autofluorescence and include single-channel controls for spectral bleed-through (standard IF practice). Because RCC1 has no transmembrane segment and its principal signal is nuclear, use permeabilisation sufficient for nuclear antibody access, checking that nuclear structure remains intact (UniProt P18754: topology and localisation; standard IF practice).
What should I check when RCC1 DAB staining looks diffuse or widespread?
Compare the section with a no-primary control to identify secondary-reagent or chromogen background before scoring RCC1 (standard IHC practice). For chromogenic detection, block endogenous peroxidase as a general workflow step and examine blood-rich, damaged, and folded regions separately (standard IHC practice). The selected image used 10% goat serum blocking, 1 μg/ml primary antibody overnight at 4°C, and DAB development; those are image-specific conditions, not a universal optimum (datasheet A02719-1). If background persists, inspect washing, blocking, primary concentration, and development time on matched sections while seeking discrete nuclear signal (standard IHC practice; HPA: general nuclear expression).
How should I score RCC1 staining across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, then score nuclear DAB within that same population across sections (standard IHC practice; HPA: general nuclear expression). An H-score combines the percentage of nuclei at intensity 0–3 and ranges from 0–300; alternatively, report the percentage of positive nuclei using a fixed threshold (standard IHC practice). Normalise counts to all evaluable nuclei in the annotated population, or report positive-cell density per mm² of viable annotated area (standard IHC practice). Keep retrieval, exposure during image capture, counterstain, and threshold settings consistent, and report mitotic figures separately if their chromosome-associated signal affects the result (UniProt P18754: localisation; standard IHC practice).
How can I distinguish true RCC1 staining from artefact?
A credible positive pattern has nuclear staining in interphase cells, with possible chromosome-associated staining in mitotic figures (UniProt P18754: localisation; HPA: general nuclear expression). Check that the signal lies in the intended cell population; HPA reports high staining in several glandular and epithelial cell populations but low staining in some glial cells and cardiomyocytes (HPA: tissue IHC). Treat staining confined to section edges, folds, or necrotic regions cautiously, and compare it with intact interior tissue (standard IHC practice). A no-primary control helps reveal endogenous-enzyme or detection-system signal; unexplained diffuse cytoplasmic DAB warrants review even though UniProt also annotates cytoplasm (UniProt P18754: localisation; standard IHC practice).
Boster reagents
Best RCC1 / Regulator of chromosome condensation IHC Antibodies
Both catalog antibodies have human paraffin-section IHC images; A02719-1 also has mouse lung IHC and human cell and tissue IF images (catalog image captions).
Real IHC data
Anti-RCC1 Antibody ®
Cat # A02719-1
Real IHC data
Anti-RCC1 Antibody ® (monoclonal, 6B11E7)
Cat # M02719-2
A02719-1 has IHC images from human rectal and lung cancer and mouse lung, plus IF images from HeLa cells and human intestine cancer tissue (A02719-1 image captions). M02719-2 has IHC images from human thyroiditis, colorectal adenocarcinoma, breast cancer and spleen, plus an IF image from A431 cells (M02719-2 image captions).
Which to pick: For human tissue IHC, both SKUs have paraffin-section images: A02719-1 is rabbit hosted, while M02719-2 is mouse monoclonal (catalog host and clone fields; respective IHC image captions). For IF/ICC, both list those applications and have human IF images; A02719-1 also lists mouse and rat reactivity and has a mouse lung IHC image, making it the better documented cross-species IHC choice (catalog applications and reactivity; respective image captions). The IHC captions identify paraffin sections but do not report the fixative for either SKU (respective IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P18754 (RCC1_HUMAN, Regulator of chromosome condensation).
- Human Protein Atlas. RCC1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. RCC1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RCC1 antibody validation summary (3 antibodies).
- Regulator of chromatin condensation 1 abrogates the G1 cell cycle checkpoint via Cdk1 in human papillomavirus E7-expressing epithelium and cervical cancer cells. Cell death & disease 2018 — PMC5964113.
- Down-regulation of RCC1 sensitizes immunotherapy by up-regulating PD-L1 via p27(kip1) /CDK4 axis in non-small cell lung cancer. Journal of cellular and molecular medicine 2021 — PMC8051708.
- RCC1 Expression as a Prognostic Marker in Colorectal Liver Oligometastases. Pathology oncology research : POR 2021 — PMC8674189.
- KRAS(G12D) mutation promotes pancreatic tumorigenesis by suppressing sirtuin three via the guanine nucleotide exchange factor RCC1. The Journal of biological chemistry 2026 — PMC12804135.
- PubMed PMID:3678831 — UniProt-cited evidence.
- PubMed PMID:1769659 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RCC1 (Regulator of chromosome condensation). Expected localisation General nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02719-1). Validated IHC antibody: A02719-1 at 0.5-1μg/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 rectal cancer tissue; fixative not specified (datasheet A02719-1); 2. Fixation: Image fixative and duration unreported (datasheet A02719-1); 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 A02719-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02719-1); 8. Primary antibody: Rabbit anti-RCC1, 0.5-1μg/ml (datasheet A02719-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02719-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Mitotic chromosome binding may change the nuclear pattern (UniProt); Isoform / epitope: Two isoforms; epitope coverage unspecified (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt P18754, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet A02719-1, PMC5964113, PMC8051708. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RCC1/immunohistochemistry-ihc-guide.