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CCNE1 / G1/S-specific cyclin-E1 · IHC design guide
Design Immunohistochemistry for CCNE1
Plan paraffin-section CCNE1 IHC using the catalog antibody at 0.5–1 μg/ml (datasheet A00543-2). Use placenta or testis as a positive tissue control and assess nuclear and cytoplasmic staining in context (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCNE1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear expected (UniProt); nuclear and cytoplasmic tissue staining observed (HPA tissue IHC) | |
| Staining pattern | Variable nuclear/cytoplasmic staining; strongest in testis and placenta (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00543-2) | |
| Positive control | Placenta+4 more · see all | |
| Negative control | Adrenal gland+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 | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended CCNE1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet: A00543-2); the two published CCNE1 IHC workflows below provide tissue microarray examples (PMC10107112; PMC4725452).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A00543-2) |
| Fixation | Image fixative and duration unreported (datasheet A00543-2); 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 A00543-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00543-2) |
| Primary antibody | Rabbit anti-CCNE1, 0.5-1μg/ml (datasheet A00543-2) |
| Primary incubation | Overnight at 4 °C (datasheet A00543-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00543-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CCNE1-positive staining in decidual cells of placenta (HPA tissue IHC: High). HPA tissue profile: Nuclear and cytoplasmic expression of varying intensity in most tissues. Highest expression in testis and placenta. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A00543-2); consider sodium citrate retrieval as a published alternative (PMC4725452).
Section 2
What Is the Expected CCNE1 Staining Pattern?
CCNE1 is a nuclear protein with no transmembrane segment (UniProt P24864: location and topology). In paraffin-section IHC, expect the strongest reported staining in placental decidual cells and cells in testicular seminiferous ducts (HPA tissue IHC: High). HPA also reports nuclear and cytoplasmic staining of varying intensity across tissues, but labels its tissue profile Approved with low consistency between antibody staining and RNA expression (HPA tissue IHC: reliability and profile).
| Distinct nuclear staining in decidual cells or cells in seminiferous ducts, with variable cytoplasmic staining. | This matches the reported High signal in these cell populations and the broader nuclear and cytoplasmic tissue pattern (HPA tissue IHC: placenta, testis, profile). Give the nuclear component particular weight because UniProt places CCNE1 in the nucleus (UniProt P24864: location). |
| Predominant membrane or extracellular staining, with no convincing nuclear signal. | Treat this as a localization mismatch: CCNE1 has no transmembrane segment and is assigned to the nucleus (UniProt P24864: topology and location). Examine the negative control and detection conditions for artefact (general IHC practice). Cytoplasmic signal alone needs a more cautious call because HPA reports it in tissue IHC (HPA tissue IHC: profile). |
| Strong staining in hepatocytes while the positive control stains as expected. | HPA reports hepatocytes as Not detected, so this pattern warrants a check for cross-reactivity or endogenous detection activity (HPA tissue IHC: liver; general IHC practice). It is not definitive proof of either cause: HPA describes low consistency between staining and RNA data (HPA tissue IHC: reliability). |
| Diffuse chromogen over nuclei, cytoplasm and surrounding tissue, obscuring cell boundaries. | The pattern cannot support compartment or cell-type scoring (general IHC practice). Check a no-primary control and the detection reagent, then review blocking and washing (general IHC practice). HPA's reported cytoplasmic expression should not be used to justify uniform background outside identifiable cells (HPA tissue IHC: profile). |
| No detectable signal in placental decidual cells or cells in testicular seminiferous ducts. | These are reported High IHC populations, so absence of staining raises a run-performance or specimen-quality question (HPA tissue IHC: placenta, testis; general IHC practice). Check the positive control, primary-antibody dilution and detection steps before scoring the study specimens as negative (general IHC practice). |
💡Expected CCNE1 appearanceCall a positive result when identifiable decidual cells or cells in seminiferous ducts show clear nuclear staining, potentially with cytoplasmic signal, at the reported High level; diffuse extracellular or dominant membrane colour is suspect (HPA tissue IHC: placenta, testis, profile; UniProt P24864: location and topology).
| Localization and topology | UniProt assigns CCNE1 to the nucleus and reports no transmembrane segment (UniProt P24864: location and topology). HPA tissue IHC also describes cytoplasmic signal; interpret that component alongside a convincing cell-associated pattern (HPA tissue IHC: profile). |
| Tissue and cell selection | Placental decidual cells and cells in testicular seminiferous ducts are High; bone-marrow hematopoietic cells and bronchial respiratory epithelial cells are Medium (HPA tissue IHC: listed populations). Select and score the named cells, not the whole section (general IHC practice). |
| Evidence strength | HPA calls the tissue staining Approved but notes low consistency with RNA expression; the listed IHC antibodies are Approved, not Enhanced (HPA tissue IHC: reliability; HPA antibodies: IHC status). Use controls and morphology when interpreting an unexpected positive or negative (general IHC practice). |
| Isoforms and epitope coverage | UniProt lists 3 CCNE1 isoforms (UniProt P24864: isoforms). The supplied record gives no antibody epitope, so it cannot establish which isoforms the IHC-validated antibody detects; check its documented epitope before making an isoform-specific interpretation (general IHC practice). |
| IF/ICC Q: What localization is reported? | A: HPA reports mainly nucleoplasmic signal and an additional, uncertain cytosolic location (HPA subcellular ICC-IF: summary and locations). This provides a localization cross-check, while the paraffin-section tissue pattern remains the basis for IHC scoring (HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive tissue is blank. | The expected High population may be absent from the section, or the IHC run may have failed (HPA tissue IHC: placenta, testis; general IHC practice). | Confirm the named cells on the counterstain; review the positive control, antigen-retrieval run, primary-antibody dilution and detection steps (general IHC practice). No CCNE1-specific retrieval condition or fixation sensitivity is established by the supplied sources. |
| Cytoplasmic signal dominates and nuclei are unclear. | HPA reports cytoplasmic tissue staining, but UniProt assigns CCNE1 to the nucleus; the result is therefore ambiguous without a clear cell-associated pattern (HPA tissue IHC: profile; UniProt P24864: location). | Compare positive and no-primary controls, review the counterstain and score nuclear and cytoplasmic compartments separately (general IHC practice). Avoid calling cytoplasmic signal alone a localization match. |
| Sharp membrane outlines or extracellular deposits dominate. | This conflicts with the nuclear assignment and lack of a transmembrane segment (UniProt P24864: location and topology); precipitated chromogen or nonspecific detection is possible (general IHC practice). | Inspect the no-primary control and reagent deposits, repeat with fresh detection reagents if needed, and require recognizable cellular staining before scoring (general IHC practice). |
| Hepatocytes stain strongly. | HPA lists hepatocytes as Not detected; cross-reactivity or endogenous detection activity is possible, though the HPA tissue profile has low RNA concordance (HPA tissue IHC: liver and reliability; general IHC practice). | Compare a no-primary control, check endogenous-activity blocking appropriate to the detection chemistry, and confirm the cell identity before assigning positivity (general IHC practice). |
| The entire section has diffuse colour. | Broad background can arise from detection chemistry, inadequate blocking or insufficient washing (general IHC practice); it does not resolve the nuclear and cytoplasmic cell pattern described by HPA (HPA tissue IHC: profile). | Check the no-primary control, blocking, washes and chromogen development; rescore only after cell boundaries and compartments are interpretable (general IHC practice). |
| Signal varies between sampled tissue areas. | HPA reports varying intensity across tissues and different levels in named cell populations (HPA tissue IHC: profile and listed populations). A change in sampled cell composition can therefore change the apparent result (general IHC practice). | Compare the same cell type across sections, document its nuclear and cytoplasmic signal separately, and keep a known-positive population in the run (general IHC practice; HPA tissue IHC: placenta and testis). |
Experiment evidence
IHC & IF images showing CCNE1 staining
A00543-2 · IHCIHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2). CCNE1 was detected in paraffin-embedded section of human placenta tissue. Heat m
A00543-2 · IHCIHC analysis of CCNE1 using anti-CCNE1 antibody (A00543-2). CCNE1 was detected in paraffin-embedded section of human placenta tissue. Heat m
M00543-4 · IHCRat colon was stained with Anti-Cyclin E1 rabbit antibody
M00543-4 · IHCHuman colon was stained with Anti-Cyclin E1 rabbit antibody
M00543 · IFImmunofluorescent analysis of Hela cells, using Cyclin E1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105173-CCNE1/tissue/cerebral+cortex#imid_5095236
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105173-CCNE1/tissue/colon#imid_4081930
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105173-CCNE1/tissue/liver#imid_5095179
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000105173-CCNE1/tissue/kidney#imid_5095180
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018169; green = CCNE1, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody HPA018169; green = CCNE1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCNE1 IHC & IF
🧪Run placenta first: decidual cells should show CCNE1 staining (HPA: High in decidual cells). Use adrenal gland glandular cells as a negative tissue (HPA: Not detected in glandular cells); on the placenta slide, cells without specific nuclear signal should remain at background, without assuming a particular cell type is negative (UniProt P24864: nucleus; standard IHC practice).
Positive control tissue: Placenta (Decidual cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CCNE1 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-matched rabbit IgG isotype controls, matching clonality where known (hero caption: rabbit primary; standard IHC practice). A CCNE1 knockout specimen can provide a biological negative; for the placenta SABC/DAB method, check endogenous peroxidase and biotin background (hero caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A00543-2 placenta caption does not state a fixative (hero caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is reported for that paraffin section, but retrieval dependency and whether frozen sections or IF are easier are unreported (hero caption: EDTA pH 8.0; supplied evidence). Placental endogenous biotin can complicate SABC detection, so interpret staining against the background controls (hero caption: SABC; standard IHC practice).
HPA tissue IHC evidence for CCNE1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCNE1 IHC Tips
Troubleshoot CCNE1 staining in paraffin sections by checking retrieval, compartment, cell type and scoring against the available IHC evidence.
How should I retrieve CCNE1 when nuclear staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A00543-2). The selected paraffin-section image used that retrieval, followed by 10% goat serum and 1 μg/ml primary antibody overnight at 4°C (datasheet A00543-2). Check whether nuclei in placenta decidual cells stain before increasing retrieval intensity, since those cells show high staining in tissue IHC (HPA: High in placenta decidual cells). If signal remains weak, compare a small retrieval-time series while keeping detection and exposure to DAB constant (standard IHC practice). Excessive heating can damage morphology and make nuclear scoring unreliable (standard IHC practice).
Could fixation explain weak CCNE1 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown because the selected tissue-IHC caption does not state a fixative (datasheet A00543-2). Record the specimen's actual fixative and fixation duration, then compare sections processed under matched conditions while using EDTA retrieval at pH 8.0 (datasheet A00543-2; standard IHC practice). Keep section thickness, primary incubation and chromogen development consistent so that fixation is the main variable being assessed (standard IHC practice). The catalog image used 1 μg/ml antibody overnight at 4°C, which supplies a starting comparison for antibody incubation (datasheet A00543-2). Do not assign a fixation effect to CCNE1 from staining distribution alone (standard IHC interpretation).
How should I assess cytoplasmic staining when CCNE1 is expected in nuclei?
Score nuclear and cytoplasmic DAB separately: CCNE1 is annotated as nuclear, while tissue IHC shows staining in both compartments (UniProt P24864 subcellular location; HPA: tissue IHC profile). Nucleoplasmic localisation has stronger subcellular support than the additional cytosolic signal, which is uncertain (HPA: subcellular localisation). Examine the hematoxylin counterstain to identify intact nuclei and reject pigment or diffuse DAB that obscures compartment boundaries (standard IHC practice). Compare the pattern with placenta decidual cells or testis cells in seminiferous ducts, where staining is high (HPA: High in placenta and testis). Report cytoplasmic staining explicitly rather than counting it as nuclear positivity (standard IHC scoring practice).
Could an isoform or modified epitope alter CCNE1 staining?
CCNE1 has 3 annotated isoforms—E1L, E1S and isoform 3—and several annotated phosphorylation sites (UniProt P24864 isoforms and modified residues). The supplied evidence does not map the catalog antibody's epitope, so it cannot establish which isoforms or phosphorylation states the antibody detects (datasheet A00543-2; UniProt P24864). When staining varies between specimens, first compare sections processed with the same EDTA retrieval at pH 8.0 and the same primary incubation (datasheet A00543-2; standard IHC practice). If isoform specificity matters to the study, obtain epitope documentation or validate against independently characterised material before assigning an isoform (standard antibody-validation practice).
How can IF help check an ambiguous CCNE1 IHC pattern?
Use IF as a separate localisation check, since the primary readout here is chromogenic IHC in paraffin sections (datasheet A00543-2). Multiplex CCNE1 with a marker identifying the expected cell population and a nuclear counterstain, then assess signal within individual nuclei; placenta decidual cells provide a tissue context with high IHC staining (HPA: High in placenta decidual cells; standard IF practice). Choose a fluorophore and filter set that separates specific signal from measured tissue autofluorescence, with an unstained section as a reference (standard IF practice). Because CCNE1 is intracellular and mainly nucleoplasmic, permeabilisation must allow antibody access across cell and nuclear membranes (UniProt P24864 subcellular location; HPA: subcellular localisation; standard IF practice).
What should I change when DAB background obscures CCNE1 staining?
Inspect a section without primary antibody alongside the stained section to locate background from secondary detection, endogenous enzyme activity or the tissue itself (standard IHC practice). Use a peroxidase block before DAB development and verify that washes remove unbound detection reagents (standard chromogenic IHC practice). The selected image used 10% goat serum, 1 μg/ml primary antibody overnight at 4°C, and a biotin-based DAB detection sequence (datasheet A00543-2). If diffuse colour persists, adjust blocking, primary concentration or chromogen development one variable at a time (standard IHC practice). Preserve nuclear morphology so background can be distinguished from plausible CCNE1 staining (UniProt P24864 subcellular location; standard IHC practice).
How should I quantify heterogeneous CCNE1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring area before reading slides, because CCNE1 staining varies by tissue and cell type (HPA: tissue IHC profile; standard IHC scoring practice). For nuclear DAB, report the percentage of positive eligible cells and an H-score calculated as 1 × % weak + 2 × % moderate + 3 × % strong (standard IHC scoring practice). If spatial abundance matters, also report positive nuclei per mm², normalised to the viable tissue area assessed (standard image-analysis practice). Keep cytoplasmic staining as a separate measure because nucleoplasmic localisation has stronger support than cytosolic localisation (HPA: subcellular localisation). Apply identical thresholds and sampling rules to every specimen (standard IHC scoring practice).
How do I distinguish credible CCNE1 positivity from staining artefact?
Give greatest weight to staining in intact nuclei of the expected cells: CCNE1 is nuclear, and placenta decidual cells and testis cells in seminiferous ducts show high tissue-IHC staining (UniProt P24864 subcellular location; HPA: tissue IHC profile). Review isolated cytoplasmic colour cautiously because the additional cytosolic localisation is uncertain (HPA: subcellular localisation). Exclude section edges, necrotic areas and pigment or DAB present in the no-primary control from positive-cell calls (standard IHC practice). Check whether peroxidase blocking removes enzyme-related signal before interpreting brown deposits as target staining (standard chromogenic IHC practice). Tissue-IHC staining has low consistency with RNA expression, so corroborate unexpected patterns independently (HPA: Approved reliability description).
Boster reagents
Best CCNE1 / G1/S-specific cyclin-E1 IHC Antibodies
Anti-CCNE1 antibodies have IHC images from human placenta and rat colon sections (A00543-2 and M00543-4 image captions), plus an IF image from HeLa cells (M00543 image caption).
Real IHC data
Anti-Cyclin E1/CCNE1 Antibody ®
Cat # A00543-2
Real IHC data
Anti-Cyclin E1 Rabbit Monoclonal Antibody
Cat # M00543-4
Real IF data
Anti-Cyclin E1 CCNE1 Rabbit Monoclonal Antibody
Cat # M00543
A00543-2 shows IHC in a paraffin-embedded human placenta section, and M00543-4 shows IHC in rat colon (respective image captions). M00543 shows IF in HeLa cells and lists IF/ICC applications (M00543 image caption; catalog applications).
Which to pick: For paraffin-section IHC, choose A00543-2: its caption specifies paraffin embedding and EDTA retrieval at pH 8.0 (A00543-2 image caption). M00543-4 is a monoclonal option for tissue IHC with human, mouse and rat listed as reactive species; its IHC image shows rat colon, while M00543 is the monoclonal choice for human IF/ICC with an IF image in HeLa cells (M00543-4 and M00543 catalog entries and image captions). Neither IHC caption reports a fixative (A00543-2 and M00543-4 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P24864 (CCNE1_HUMAN, G1/S-specific cyclin-E1).
- Human Protein Atlas. CCNE1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CCNE1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. CCNE1 antibody validation summary (3 antibodies).
- Comparative Analysis of Cyclin E1 Expression in Uterine Leiomyosarcomas and Leiomyomas. Diseases (Basel, Switzerland) 2026 — PMC13604967.
- CCNE1 expression in high grade serous carcinoma does not correlate with chemoresistance. Oncotarget 2017 — PMC5617501.
- CCNE1 and survival of patients with tubo-ovarian high-grade serous carcinoma: An Ovarian Tumor Tissue Analysis consortium study. Cancer 2023 — PMC10107112.
- CCNE1 amplification is associated with aggressive potential in endometrioid endometrial carcinomas. International journal of oncology 2016 — PMC4725452.
- PubMed PMID:1833066 — UniProt-cited evidence.
- PubMed PMID:1833068 — UniProt-cited evidence.
- PubMed PMID:7739542 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCNE1 (G1/S-specific cyclin-E1). Expected localisation Nuclear expected (UniProt); nuclear and cytoplasmic tissue staining observed (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00543-2). Validated IHC antibody: A00543-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Placenta (Decidual cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A00543-2); 2. Fixation: Image fixative and duration unreported (datasheet A00543-2); 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 A00543-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00543-2); 8. Primary antibody: Rabbit anti-CCNE1, 0.5-1μg/ml (datasheet A00543-2); 9. Primary incubation: Overnight at 4 °C (datasheet A00543-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt P24864, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A00543-2, PMC10107112, PMC4725452. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCNE1/immunohistochemistry-ihc-guide.