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RCOR1 / REST corepressor 1 · IHC design guide
Design Immunohistochemistry for RCOR1
Plan RCOR1 paraffin IHC around its general nuclear staining pattern (HPA tissue IHC). Compare high staining in bone marrow hematopoietic cells with low staining in parathyroid glandular cells (HPA tissue IHC), using 2–5 μg/ml as the catalog antibody's starting range (datasheet A02799-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
RCOR1 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 | Predominantly nuclear in tissue sections (HPA tissue IHC) | |
| Staining pattern | General nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02799-3) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Parathyroid glandular cells show low staining (HPA tissue IHC) | |
| Regulation | No induction specified in the record (UniProt) | |
| Isoform / epitope | No isoforms annotated; one 1–485 chain (UniProt) |
Section 1
Recommended RCOR1 IHC & IF Protocols
The catalog antibody IHC-P protocol (datasheet A02799-3) is paired with a published RCOR1 protocol for HCC tissue microarrays (PMC10277624).
| Sample | Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A02799-3) |
| Fixation | Image fixative and duration unreported (datasheet A02799-3); 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 A02799-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02799-3) |
| Primary antibody | Rabbit anti-RCOR1, 2-5 μg/ml (datasheet A02799-3) |
| Primary incubation | Overnight at 4 °C (datasheet A02799-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02799-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | RCOR1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 (datasheet A02799-3). Tris/EDTA pH 9.0 is a published alternative used with a different RCOR1 antibody (PMC10277624).
Section 2
What Is the Expected RCOR1 Staining Pattern?
RCOR1 should stain nuclei across many cell types: HPA reports general nuclear expression, high staining in several listed tissues, and low tissue specificity (HPA tissue IHC: Supported). Its nuclear localisation and lack of a transmembrane segment support a nuclear reading of the stain (UniProt Q9UKL0: subcellular location; topology). Compare compartments and cell populations within each section, since broadly distributed expression does not imply equal intensity in every cell (HPA tissue IHC).
| Nuclear chromogen appears in hematopoietic cells of bone marrow and glandular cells of adrenal gland or appendix (HPA tissue IHC: High). | This fits the reported general nuclear pattern; judge signal in identifiable cells against the counterstain and background, allowing intensity to vary between cells (HPA tissue IHC: Supported; General IHC practice). |
| Cytoplasmic or membranous staining dominates while nuclei remain pale. | Treat this as discordant with the expected nuclear pattern and review staining specificity and detection controls (UniProt Q9UKL0: nucleus; HPA tissue IHC: general nuclear expression). UniProt reports partial cytoplasmic translocation during HSV-1 infection, so infection context matters before assigning artefact (UniProt Q9UKL0: subcellular location). |
| Strong chromogen marks an unexpected cell population or acellular material, without matching nuclei. | Check for cross-reactivity or endogenous detection activity before calling it RCOR1 (General IHC practice). HPA reports broad expression, so cell identity alone cannot establish a negative population; compare its compartment and an appropriate control (HPA tissue IHC: general nuclear expression; General IHC practice). |
| Diffuse colour fills stroma, lumina, or most of the section and obscures nuclear boundaries. | This is background rather than interpretable nuclear localisation (General IHC practice). Review blocking, reagent concentration, detection chemistry, and washes using a control section; HPA's tissue profile does not identify the procedural cause (HPA tissue IHC; General IHC practice). |
| No nuclear signal is visible in hematopoietic cells of bone marrow or adipocytes of adipose tissue (HPA tissue IHC: High). | A missing signal in these reported high-staining populations calls for a run check before interpreting other tissue as negative (HPA tissue IHC; General IHC practice). Inspect control performance, antibody use, retrieval, and detection; the supplied sources do not establish RCOR1-specific fixation sensitivity. |
💡Expected RCOR1 appearanceCall a result consistent when identifiable nuclei show distinct chromogen, including high signal in HPA-listed bone marrow hematopoietic cells or adipose adipocytes (HPA tissue IHC: High; general nuclear expression); dominant cytoplasmic colour or diffuse noncellular deposits are suspect, subject to the reported HSV-1 exception (UniProt Q9UKL0: subcellular location; General IHC practice).
| Reference tissue and cell population | HPA reports high staining in adipocytes of adipose tissue and breast; glandular cells of adrenal gland and appendix; marrow hematopoietic cells; bronchial respiratory epithelial cells; caudate glial cells; and cortical endothelial cells (HPA tissue IHC: High). Parathyroid glandular cells are reported low, so use a listed high-staining population for a positive run check (HPA tissue IHC: Low; General IHC practice). |
| Antibody evidence | HPA lists IHC support for HPA054241 and CAB001956; HPA049327 has no listed IHC status (HPA antibodies). The tissue profile is Supported because antibody staining is consistent with RNA expression; this does not validate every antibody or every protocol (HPA tissue IHC: reliability description; HPA antibodies). |
| Compartment and protein context | RCOR1 is nuclear, has no transmembrane segment or signal peptide, and is recorded as one chain of residues 1–485 (UniProt Q9UKL0: subcellular location; topology; processing). These facts support evaluating nuclear staining; they do not predict retrieval conditions or a fixation response. |
| Modified residues and epitope limits | UniProt records phosphoserines at positions 127, 260, and 460 and no glycosylation sites (UniProt Q9UKL0: modified residues; glycosylation). Their presence alone does not show whether a particular antibody's epitope or IHC signal changes; no epitope location is supplied. |
| IF/ICC Q&A: Where should RCOR1 appear? | In the nucleoplasm: HPA calls this its enhanced main location and lists ICC-IF images in A-431, MCF-7, U2OS, and NIH 3T3 (HPA subcellular). That imaging evidence supports a localisation comparison, while the primary interpretation here concerns paraffin-section IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| A listed high-staining control tissue has no nuclear chromogen (HPA tissue IHC: High). | An unsuccessful staining run or weak detection is possible; the tissue result alone does not identify the failed step (General IHC practice). | Review the control section, catalog antibody instructions, retrieval run, reagent activity, and chromogenic detection before scoring samples (General IHC practice). Do not infer RCOR1-specific fixation sensitivity from this result. |
| Nuclei are pale, but cytoplasm is strongly stained. | The compartment conflicts with the usual pattern (HPA tissue IHC: general nuclear expression; UniProt Q9UKL0: nucleus). HSV-1 infection is a documented exception (UniProt Q9UKL0: subcellular location). | Check infection context, antibody specificity evidence, and control sections; call the cytoplasmic signal unexplained if the exception is inapplicable (UniProt Q9UKL0: subcellular location; General IHC practice). |
| Colour appears in an unexpected cell population or on acellular material. | Cross-reactivity or endogenous detection activity may contribute (General IHC practice). Broad RCOR1 expression means an unexpected cell identity alone is inconclusive (HPA tissue IHC: low tissue specificity). | Compare nuclear localisation with a listed high-staining control; use appropriate reagent and detection controls to test background before assigning cellular positivity (HPA tissue IHC: general nuclear expression; General IHC practice). |
| The section has diffuse colour that hides nuclear detail. | Nonspecific reagent binding or excessive chromogenic background may obscure the expected pattern (General IHC practice; HPA tissue IHC: general nuclear expression). | Review blocking, antibody concentration against its supplied instructions, washes, and detection timing with a control section (General IHC practice). Avoid assigning a target-specific cause without supporting evidence. |
| Parathyroid glandular cells stain less than a positive control. | Low staining in that population is reported by HPA; lower intensity there need not indicate run failure (HPA tissue IHC: Low). | Judge run performance in a listed high-staining population, then assess parathyroid nuclei against local background and counterstain (HPA tissue IHC: High; General IHC practice). |
| Different antibody preparations give conflicting nuclear patterns. | Their IHC evidence differs: HPA054241 and CAB001956 are IHC Supported, while HPA049327 has no listed IHC status (HPA antibodies). | Review each antibody's stated IHC validation and instructions, then compare staining in the same listed high-staining cell population and with the same scoring criteria (HPA antibodies; HPA tissue IHC: High; General IHC practice). |
Experiment evidence
IHC & IF images showing RCOR1 staining
A02799-3 · IHCIHC analysis of CoREST/RCOR1 using anti-CoREST/RCOR1 antibody (A02799-3). CoREST/RCOR1 was detected in a paraffin-embedded section of human
A02799-3 · IFIF analysis of CoREST/RCOR1 using anti-CoREST/RCOR1 antibody (A02799-3). CoREST/RCOR1 was detected in an immunocytochemical section of SiHa
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000089902-RCOR1/tissue/cerebral+cortex#imid_641496
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000089902-RCOR1/tissue/colon#imid_641438
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000089902-RCOR1/tissue/liver#imid_641431
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000089902-RCOR1/tissue/kidney#imid_641424
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049327; green = RCOR1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA049327; green = RCOR1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for RCOR1 IHC & IF
🧪Run bone marrow first: hematopoietic cells have High RCOR1 staining (HPA: bone marrow, hematopoietic cells), with nuclear signal expected (UniProt Q9UKL0: nucleus). HPA detects RCOR1 in all 44 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells without nuclear staining on the positive slide as observations rather than validated internal negatives (HPA: no negative rows; standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: RCOR1 is detected in all 44 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 RCOR1 in A-431, MCF-7, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a matched nonimmune rabbit IgG isotype control, and an RCOR1 knockout specimen processed alongside the positive slide as a biological negative (selected-SKU caption: rabbit primary; standard IHC practice). In bone marrow, block endogenous peroxidase before DAB detection and check for endogenous biotin when using the caption’s streptavidin–biotin system (selected-SKU caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0, so start there and assess whether nuclear staining is retrieval dependent (selected-SKU caption; UniProt Q9UKL0: nucleus). The caption does not state a fixative, and the supplied evidence reports no RCOR1-specific fixation window or fixation effect (selected-SKU caption: fixative unreported). Frozen sections are not established as easier; IF can assess nucleoplasmic localization in the listed imaged cell lines, while bone marrow requires attention to endogenous peroxidase in chromogenic IHC (HPA subcellular: nucleoplasm and ICC-IF images; standard IHC practice).
HPA tissue IHC evidence for RCOR1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: RCOR1 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced RCOR1 IHC Tips
Troubleshoot RCOR1 staining in paraffin sections by checking retrieval, nuclear localisation, tissue controls and scoring consistency before interpreting chromogenic signal.
Which retrieval condition should I use when nuclear RCOR1 staining is weak?
Start with heat-mediated EDTA at pH 8.0 for this catalog antibody (datasheet A02799-3). The documented paraffin-section example used that retrieval condition before overnight primary incubation at 4°C, but did not report a heating time (IHC caption A02799-3). If nuclei remain faint, compare a longer or shorter heating interval on matched sections while keeping antibody concentration, detection and development constant; an alternative buffer should be a subsequent, separately controlled trial. Score whether nuclear contrast improves without tissue damage or diffuse staining, using a known positive section in every run (HPA tissue IHC: general nuclear expression).
How should I troubleshoot suspected fixation effects on RCOR1 staining?
The selected paraffin-section caption does not state its fixative, so target-specific RCOR1 fixation sensitivity is unknown (IHC caption A02799-3). Record the fixative, fixation duration and processing history for each specimen, then compare sections processed together before changing retrieval or antibody concentration. Uneven staining across a section can reflect processing variation; inspect morphology and compare nuclei in intact areas with those near damaged edges. If fixation histories differ, assess each batch with its own positive control and treat any staining difference as provisional until a matched processing comparison supports it.
What staining pattern should count as RCOR1 positive in paraffin sections?
Prioritise nuclear staining: RCOR1 is annotated in the nucleus, and the tissue IHC profile describes general nuclear expression (UniProt Q9UKL0 subcellular location; HPA tissue IHC). More specifically, the subcellular dataset places RCOR1 in the nucleoplasm, so compare chromogen with the nuclear counterstain at single-cell resolution (HPA subcellular: nucleoplasm). Score cytoplasmic-only colour separately and review detection background before calling those cells positive. Cytoplasmic RCOR1 can occur during HSV-1 infection, but that conditional annotation alone does not establish the cause of cytoplasmic staining in an uncharacterised section (UniProt Q9UKL0 subcellular location).
Could epitope location or modification explain inconsistent RCOR1 staining?
The supplied record lists 0 isoforms, so an isoform switch is unsupported as an explanation here (UniProt Q9UKL0 isoforms). RCOR1 has no transmembrane segment and contains ELM2 and two SANT domains, while annotated phosphoserines include residues 127, 260 and 460 (UniProt Q9UKL0 topology, domains and modified residues). The selected caption does not identify the antibody epitope, so do not assign a staining difference to any one domain or phosphorylation site (IHC caption A02799-3). Compare retrieval conditions and matched controls first; an epitope-specific explanation requires independent mapping or validation.
How can IF help resolve ambiguous nuclear staining seen by chromogenic IHC?
Use IF as a separate localisation check: RCOR1 is reported in the nucleoplasm, while the tissue IHC profile describes general nuclear expression (HPA subcellular; HPA tissue IHC). Multiplex with a marker for the cell population being assessed and a nuclear counterstain, then inspect whether the RCOR1 channel overlaps nuclei within that population. Choose a fluorophore channel with low tissue autofluorescence and include single-channel controls to evaluate bleed-through. For this protein, which has no transmembrane segment, use controlled permeabilisation to give antibody access to the nuclear epitope; optimise that step for the IF specimen independently of the paraffin-section method (UniProt Q9UKL0 topology; IHC caption A02799-3).
How do I distinguish RCOR1 signal from chromogenic background?
Begin with an intact positive section and a no-primary control, then compare nuclear signal against extracellular colour, folds and section edges. The documented workflow used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a biotinylated secondary for 30 minutes at 37°C, and DAB development (IHC caption A02799-3). Treat peroxidase blocking and careful DAB timing as general chromogenic workflow controls, and assess nonspecific secondary or detection-system staining in controls. Retain only cell-associated nuclear colour above the local background when scoring, consistent with the reported RCOR1 tissue pattern (HPA tissue IHC: general nuclear expression).
What is a defensible way to score RCOR1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable analysis area before scoring, because the reference tissue profile describes broad nuclear expression (HPA tissue IHC). Record the percentage of positive nuclei and staining intensity, then calculate an H-score as the sum of each intensity category’s percentage multiplied by its category value. Alternatively, report positive-nucleus density per mm² when cell abundance varies; normalise that count to the viable area examined and also report total nuclei. Apply one threshold, counterstain assessment and image-acquisition setting across comparison groups, with blinded review of borderline nuclei.
When should an apparent RCOR1-positive field be treated as artefactual?
A convincing result shows cell-associated nuclear staining in morphologically intact tissue, consistent with RCOR1’s nuclear annotation and the supported general nuclear IHC profile (UniProt Q9UKL0 subcellular location; HPA tissue IHC). Question cytoplasmic-only colour, staining confined to section edges or necrotic areas, and signal that also appears in a no-primary control. Review the peroxidase block and detection controls if chromogen follows endogenous enzyme activity rather than nuclei. Interpret cell-type differences against the tissue context: HPA reports high staining in several sampled populations and low staining in parathyroid glandular cells, so a single universal intensity cutoff may mislead (HPA tissue IHC).
Boster reagents
Best RCOR1 / REST corepressor 1 IHC Antibodies
A02799-3 has IHC data from a human paraffin-embedded tissue section and IF/ICC data from SiHa cells (catalog image captions); its listed reactivity is human (catalog applications/reactivity).
Real IHC data
Anti-CoREST/RCOR1 Antibody ®
Cat # A02799-3
A02799-3 is the SKU that will render with an IHC figure from a human esophageal squamous carcinoma paraffin section (catalog IHC image caption). Its IF/ICC figure shows SiHa cells (catalog IF image caption), and its listed reactivity is human (catalog reactivity).
Which to pick: Choose A02799-3 for human tissue IHC: its own image caption documents staining in a paraffin section, with the fixative unreported (catalog IHC image caption). Choose A02799-3 for IF/ICC when image-backed evidence matters: its own caption documents SiHa cells (catalog IF image caption). For mouse or rat work, M02799-1 is the listed rabbit monoclonal option with IHC and IF/ICC applications, but the payload supplies no IHC or IF image for it (catalog clone, reactivity, applications and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UKL0 (RCOR1_HUMAN, REST corepressor 1).
- Human Protein Atlas. RCOR1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. RCOR1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. RCOR1 antibody validation summary (3 antibodies).
- HKDC1 promotes colorectal cancer progression by regulating RCOR1 expression to activate the Wnt/β-catenin pathway, enhancing proliferation, migration, and epithelial-mesenchymal transition. The Journal of biological chemistry 2025 — PMC12137164.
- Recurrent GRHL fusions in a subset of sebaceoma: microscopic and molecular characterisation of eight cases. Histopathology 2025 — PMC11791738.
- Comprehensive analysis of REST corepressors (RCORs) in pan-cancer. Frontiers in cell and developmental biology 2023 — PMC10277624.
- OVOL2 sustains postnatal thymic epithelial cell identity. Nature communications 2023 — PMC10682436.
- PubMed PMID:12508121 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:10449787 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for RCOR1 (REST corepressor 1). Expected localisation Predominantly nuclear in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02799-3). Validated IHC antibody: A02799-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 44 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet A02799-3); 2. Fixation: Image fixative and duration unreported (datasheet A02799-3); 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 A02799-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02799-3); 8. Primary antibody: Rabbit anti-RCOR1, 2-5 μg/ml (datasheet A02799-3); 9. Primary incubation: Overnight at 4 °C (datasheet A02799-3). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Parathyroid glandular cells show low staining (HPA tissue IHC); Isoform / epitope: No isoforms annotated; one 1–485 chain (UniProt); Regulation: No induction specified in the record (UniProt). Evidence: UniProt Q9UKL0, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A02799-3, PMC10277624. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/RCOR1/immunohistochemistry-ihc-guide.