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RB1CC1 / RB1-inducible coiled-coil protein 1 · IHC design guide
Design Immunohistochemistry for RB1CC1
Plan RB1CC1 chromogenic IHC around cytoplasmic staining in most tissues, including epithelial cells (HPA tissue IHC). Start with the catalog antibody's 2–5 μg/ml IHC range (datasheet A02414-1) and assess staining with appropriate controls (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RB1CC1 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues, including epithelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02414-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Bone marrow+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 has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression correlates with RB1 levels (UniProt) | |
| Isoform / epitope | Two isoforms; confirm epitope coverage for both (UniProt) |
Section 1
Recommended RB1CC1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by three published RB1CC1 protocols using human breast cancer sections (PMC2894861; PMC3008740; PMC3291565).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet A02414-1) |
| Fixation | Image fixative and duration unreported (datasheet A02414-1); 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 A02414-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02414-1) |
| Primary antibody | Rabbit anti-RB1CC1, 2-5 μg/ml (datasheet A02414-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02414-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02414-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RB1CC1-positive staining in ciliated cells (cilia axoneme) of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, including epithelial cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A02414-1); the published protocols used autoclaving or another heating method (PMC2894861; PMC3008740; PMC3291565).
Section 2
What Is the Expected RB1CC1 Staining Pattern?
Expect predominantly cytoplasmic RB1CC1 staining across many tissues, including epithelial cells (HPA tissue IHC). Ciliated cells in bronchus, fallopian tube and nasopharynx show High axonemal staining, while several glandular cell populations show Medium staining (HPA tissue IHC). RB1CC1 has no transmembrane segment (UniProt Q8TDY2 topology). Interpret tissue patterns with caution: HPA rates its IHC evidence Approved, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in epithelial cells, with strong ciliary axoneme staining in bronchus or fallopian tube (HPA tissue IHC). | This fits the reported tissue pattern: cytoplasmic expression in most tissues and High signal in those ciliated cells (HPA tissue IHC). Score the cell compartment and ciliary signal separately so a strong axoneme does not imply uniformly strong cytoplasm. |
| Predominantly nuclear-only staining, without the expected cytoplasmic pattern (HPA tissue IHC). | Treat this as a discordant result and examine controls and detection conditions. UniProt also lists the nucleus, so nuclear signal alone cannot prove an artifact (UniProt Q8TDY2 subcellular location); its dominance still differs from HPA's usual tissue pattern (HPA tissue IHC). |
| Strong signal in a cell population HPA reports as Not detected, such as bone-marrow hematopoietic cells (HPA tissue IHC). | Investigate cross-reactivity or endogenous detection activity before scoring it as RB1CC1. An HPA Not detected result is a comparator for that cell population, not proof that every specimen must be negative (HPA tissue IHC). |
| Diffuse chromogen covers tissue, extracellular space and regions lacking recognizable cell boundaries. | This prevents reliable cytoplasmic or ciliary scoring. Check the negative detection control and review blocking, washes and chromogen development as general IHC practice; the supplied HPA and UniProt records do not identify a target-specific cause. |
| No signal in bronchial or fallopian-tube ciliated cells, where HPA reports High staining (HPA tissue IHC). | A negative run there calls for a technical review before interpreting other tissues as negative. Check section integrity, primary antibody, detection and counterstain using routine IHC controls; HPA's Approved rating still allows specimen-to-specimen variation (HPA tissue IHC). |
💡Expected RB1CC1 appearanceA convincing positive shows identifiable cytoplasmic staining, with High ciliary axoneme signal in the specified ciliated epithelia or Medium signal in reported glandular cells (HPA tissue IHC); broad extracellular or control-section color is suspect by standard IHC interpretation.
| Cell and compartment choice | HPA reports cytoplasmic expression in most tissues, including epithelial cells, but High signal specifically at ciliary axonemes in bronchus, fallopian tube and nasopharynx (HPA tissue IHC). Document which structure was scored rather than pooling all visible signal. |
| Strength of tissue evidence | The tissue profile has Approved reliability and medium consistency with RNA expression (HPA tissue IHC). Both listed antibodies have IHC Approved status (HPA antibodies). Use the profile as a pattern reference, not an absolute pass/fail threshold for an individual section. |
| What should IF/ICC show? | Mainly cytosolic signal is supported; nuclear-membrane localisation is uncertain (HPA subcellular ICC-IF). Only HPA053049 has ICC Supported status among the listed antibodies (HPA antibodies). IF/ICC localisation is context for interpretation, not an IHC-P protocol option. |
| Isoforms and antibody epitope | UniProt lists 2 isoforms (UniProt Q8TDY2 isoforms). The supplied records do not locate the catalog antibody's epitope or establish isoform-specific staining. Avoid assigning different cell patterns to isoforms from these data alone. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive ciliated epithelium is blank (HPA tissue IHC). | The run may have failed at primary incubation, detection or chromogen development; the records do not identify which step. | Review the IHC-validated antibody's IHC-P instructions and run a known-positive section alongside a negative detection control. Confirm cilia are present on the section before treating absent axonemal color as biological. |
| Only nuclei stain intensely while epithelial cytoplasm is blank (HPA tissue IHC). | The pattern conflicts with HPA's predominant cytoplasmic profile, although UniProt also lists nuclear localisation (HPA tissue IHC; UniProt Q8TDY2 subcellular location). | Compare with a known-positive tissue and a negative detection control. Record nuclear and cytoplasmic scores separately; do not call all nuclear color false solely from the HPA pattern. |
| Bone-marrow hematopoietic cells or heart cardiomyocytes stain strongly despite HPA Not detected entries (HPA tissue IHC). | Cross-reactivity or endogenous detection activity is possible; a discordant specimen alone cannot distinguish them. | Inspect the negative detection control and cellular morphology. Recheck the pattern with an independently validated antibody when available, then report the discrepancy against the specific HPA cell population. |
| Brown deposit appears throughout the section, including spaces between cells. | Nonspecific background or endogenous detection activity can obscure cell-associated staining under standard chromogenic IHC practice. | Compare a negative detection control; review blocking, wash steps and chromogen development. Score RB1CC1 only where the signal follows identifiable cellular structures. |
| A weak glandular section is called negative because it lacks the ciliary intensity. | The comparator is mismatched: HPA reports Medium glandular staining in several tissues and High axonemal staining in specified ciliated cells (HPA tissue IHC). | Score each reported cell type and structure against its own HPA level. Include a positive reference section in the run and document intensity and localisation separately. |
| Puncta are interpreted as a required RB1CC1 pattern in routine tissue IHC. | UniProt describes punctate preautophagosomal structures under starvation, while HPA's general tissue IHC profile emphasizes cytoplasmic expression (UniProt Q8TDY2 subcellular location; HPA tissue IHC). | Record puncta if present, but do not require them for a positive tissue result. Confirm the experimental context before linking puncta to starvation-associated localisation. |
Experiment evidence
IHC & IF images showing RB1CC1 staining
A02414-1 · IHCIHC analysis of RB1CC1 using anti-RB1CC1 antibody (A02414-1). RB1CC1 was detected in a paraffin-embedded section of human colon tissue. Heat
A02414-1 · IHCIHC analysis of RB1CC1 using anti-RB1CC1 antibody (A02414-1). RB1CC1 was detected in a paraffin-embedded section of human colon cancer tissu
A02414-1 · IFIF analysis of RB1CC1 using anti-RB1CC1 antibody (A02414-1). RB1CC1 was detected in an immunocytochemical section of U2OS cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000023287-RB1CC1/tissue/cerebral+cortex#imid_15494407
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000023287-RB1CC1/tissue/colon#imid_15494325
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000023287-RB1CC1/tissue/liver#imid_15494318
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000023287-RB1CC1/tissue/kidney#imid_15494319
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA053049; green = RB1CC1, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA053049; green = RB1CC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RB1CC1 IHC & IF
🧪Run bronchus first: ciliated cells should show ciliary axoneme staining (HPA: High in bronchus ciliated cells). Use heart muscle as the negative tissue because cardiomyocytes are listed as not detected (HPA: heart muscle); on the bronchus slide, adjacent non-ciliated cells should lack the ciliary axoneme pattern, though they are not a validated RB1CC1-negative population.
Positive control tissue: Bronchus (Ciliated cells (cilia axoneme), HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RB1CC1 in HEK293, PC-3, U2OS, NIH 3T3, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality where applicable (caption: rabbit anti-RB1CC1); use matched RB1CC1 knockout material as a biological specificity control. Quench endogenous peroxidase for chromogenic detection and check the bronchial ciliary edge for background staining (caption: HRP/DAB detection; HPA: bronchus cilia axoneme staining).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative for the selected A02414-1 paraffin-section caption is unreported (caption: fixative not stated). That caption provides an IHC retrieval starting point of EDTA at pH 8.0, but the strong bronchial cilia pattern warrants checking for ciliary-edge background (caption: heat-mediated EDTA retrieval; HPA: High in bronchus cilia axoneme). Frozen sections are not established as easier; IF/ICC has separate guidance, with supported cytosolic localization in the supplied imaging evidence (HPA: cytosol supported).
HPA tissue IHC evidence for RB1CC1
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 |
|---|---|---|---|---|
| Bronchus | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (cilia axoneme) | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced RB1CC1 IHC Tips
Troubleshoot RB1CC1 staining by comparing matched paraffin sections and judging signal against documented cell types and compartments (datasheet A02414-1; HPA tissue IHC; HPA subcellular).
Which retrieval condition should I start with for RB1CC1 paraffin-section IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 for RB1CC1 paraffin-section IHC (datasheet A02414-1). The selected image used a 10% goat-serum block and 2 μg/ml primary antibody overnight at 4°C, giving a matched baseline before changing retrieval (datasheet A02414-1). If staining is weak, compare retrieval durations on adjacent sections while holding antibody concentration and DAB development constant, and include a no-primary control (standard IHC practice). Favor conditions that reveal glandular-cell staining in colon without disproportionate edge staining or diffuse nuclear color (HPA tissue IHC: medium in colon glandular cells; standard IHC practice).
How should I investigate variable RB1CC1 staining across paraffin blocks?
RB1CC1-specific sensitivity to fixation is unknown because the selected paraffin-section caption does not state a fixative (datasheet A02414-1). Record the actual fixative, fixation duration, processing run and section age for each block before comparing staining (standard IHC practice). On adjacent sections, keep EDTA retrieval at pH 8.0 and start with the caption's 2 μg/ml primary incubation overnight at 4°C (datasheet A02414-1). Compare morphology, controls and cytoplasmic staining together; differences between blocks alone cannot establish an RB1CC1-specific fixation effect (HPA tissue IHC: predominantly cytoplasmic expression; standard IHC practice).
Where should convincing RB1CC1 chromogenic staining appear?
Expect a predominantly cytoplasmic pattern across many tissues, with supported cytosolic localisation in cell imaging (HPA tissue IHC; HPA subcellular: cytosol supported). Nuclear, lysosomal and preautophagosomal localisations are also annotated, while nuclear-membrane localisation has uncertain support (UniProt Q8TDY2 localisation; HPA subcellular). Review glandular cells in colon as a reference for medium tissue staining rather than requiring every cell to stain equally (HPA tissue IHC: medium in colon glandular cells). If DAB appears exclusively nuclear, compare adjacent sections, a no-primary control and cell morphology before assigning it to RB1CC1 (standard IHC practice; HPA tissue IHC: predominantly cytoplasmic).
Could isoforms or modified residues explain a discordant RB1CC1 IHC pattern?
RB1CC1 has 2 annotated isoforms and 19 modified residues, including phosphorylation at Ser222 and Ser229 (UniProt Q8TDY2). The supplied antibody caption gives no epitope position, so its isoform coverage and sensitivity to those modifications cannot be assigned (datasheet A02414-1). If staining conflicts with the expected cell pattern, check the antibody's documented immunogen or epitope and compare an independently mapped antibody on adjacent sections when available (standard IHC practice; HPA tissue IHC). Keep EDTA retrieval at pH 8.0 during that comparison so a retrieval change does not masquerade as epitope dependence (datasheet A02414-1; standard IHC practice).
How can IF help assess an ambiguous RB1CC1 IHC pattern?
Use IF as a separate localisation check, multiplexing RB1CC1 with a marker for the expected cell type; colon glandular cells provide one documented tissue context (HPA tissue IHC: medium in colon glandular cells; standard IF practice). Choose fluorophores after inspecting tissue autofluorescence in unstained controls, and separate the marker and RB1CC1 emission channels (standard IF practice). RB1CC1 has no transmembrane segment, but its annotated cytosolic, nuclear and organelle-associated locations do not establish where this antibody binds (UniProt Q8TDY2 topology and localisation; datasheet A02414-1). Set permeabilisation for the mapped epitope's accessible side and validate that choice in IF; do not transfer the paraffin-section EDTA pH 8.0 condition directly (standard IF practice; datasheet A02414-1).
How do I distinguish RB1CC1 signal from chromogenic background?
The selected IHC image used a 10% goat-serum block, a rabbit primary at 2 μg/ml and peroxidase-based DAB detection (datasheet A02414-1). Run a no-primary section and apply a peroxidase block to assess secondary-reagent binding and endogenous enzyme activity (standard IHC practice). If background persists, compare shorter DAB development or a primary-antibody titration on adjacent sections while keeping EDTA retrieval at pH 8.0 (standard IHC practice; datasheet A02414-1). Judge improvement by preservation of cell-associated cytoplasmic staining, especially in documented colon glandular cells, rather than by a uniformly paler section (HPA tissue IHC: medium in colon glandular cells; standard IHC practice).
What should I score when comparing RB1CC1 IHC between sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since tissue IHC is predominantly cytoplasmic and colon glandular cells have documented medium staining (HPA tissue IHC). Score cytoplasmic intensity with an H-score from 0–300, or report the percentage of positive eligible cells using a fixed threshold (standard IHC practice). Normalise positive counts to all evaluable cells of the same type, or positive area to measured viable tissue area, rather than to the whole slide (standard IHC practice). Keep retrieval at EDTA pH 8.0, DAB development, imaging and exclusion rules consistent; document necrotic and edge regions separately (datasheet A02414-1; standard IHC practice).
When is an apparent RB1CC1-positive region likely to be artefactual?
A credible result follows cell morphology and the predominantly cytoplasmic tissue pattern, with medium staining documented in colon glandular cells (HPA tissue IHC). Strong DAB limited to section edges, folds or necrotic areas warrants review on adjacent sections because those patterns can reflect processing artefact (standard IHC practice). An exclusively nuclear result needs additional validation: nuclear localisation is annotated, but supported cell imaging places RB1CC1 mainly in the cytosol (UniProt Q8TDY2 localisation; HPA subcellular). Compare no-primary and peroxidase-blocked controls, and check whether staining occurs in the expected cell type before calling an enzyme deposit or isolated dark cell RB1CC1-positive (standard IHC practice; HPA tissue IHC).
Boster reagents
Best RB1CC1 / RB1-inducible coiled-coil protein 1 IHC Antibodies
A02414-1 has real IHC data from paraffin-embedded human colon, colon cancer, breast cancer, and mouse heart sections, plus IF data from U2OS cells and human breast cancer sections (catalog image captions).
Real IHC data
Anti-RB1CC1 Antibody ®
Cat # A02414-1
A02414-1 has IHC images from paraffin-embedded human colon, colon cancer, and breast cancer sections and mouse heart sections (A02414-1 IHC image captions). Its IF images show U2OS cells and paraffin-embedded human breast cancer sections; the catalog lists human, mouse, and rat reactivity (A02414-1 IF image captions; catalog reactivity).
Which to pick: Choose A02414-1 for chromogenic tissue IHC: its paraffin-section captions specify EDTA retrieval at pH 8.0, 2 μg/ml primary antibody, and peroxidase/DAB detection; the fixative is unreported (A02414-1 IHC image captions). For IF/ICC, A02414-1 is listed for both applications and has IF images from U2OS cells and human breast cancer sections at 5 μg/ml (catalog applications; A02414-1 IF image captions). For cross-species IHC, A02414-1 lists human, mouse, and rat reactivity, with IHC images from human and mouse sections; its host is rabbit and clonality is unreported (catalog reactivity and host; A02414-1 IHC image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TDY2 (RBCC1_HUMAN, RB1-inducible coiled-coil protein 1).
- Human Protein Atlas. RB1CC1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. RB1CC1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nuclear membrane..
- Human Protein Atlas. RB1CC1 antibody validation summary (2 antibodies).
- RB1CC1 activates RB1 pathway and inhibits proliferation and cologenic survival in human cancer. PloS one 2010 — PMC2894861.
- RB1CC1 together with RB1 and p53 predicts long-term survival in Japanese breast cancer patients. PloS one 2010 — PMC3008740.
- Preparation of mouse monoclonal antibody for RB1CC1 and its clinical application. PloS one 2012 — PMC3291565.
- Comparison of the expression of TGF-β1, E-cadherin, N-cadherin, TP53, RB1CC1 and HIF-1α in oral squamous cell carcinoma and lymph node metastases of humans and mice. Oncology letters 2018 — PMC5774522.
- PubMed PMID:11850849 — UniProt-cited evidence.
- PubMed PMID:9039502 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RB1CC1 (RB1-inducible coiled-coil protein 1). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02414-1). Validated IHC antibody: A02414-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (cilia axoneme)); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet A02414-1); 2. Fixation: Image fixative and duration unreported (datasheet A02414-1); 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 A02414-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02414-1); 8. Primary antibody: Rabbit anti-RB1CC1, 2-5 μg/ml (datasheet A02414-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02414-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: Two isoforms; confirm epitope coverage for both (UniProt); Regulation: Expression correlates with RB1 levels (UniProt). Evidence: UniProt Q8TDY2, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A02414-1, PMC2894861, PMC3008740. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RB1CC1/immunohistochemistry-ihc-guide.