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CCNE1 / G1/S-specific cyclin-E1 · IHC design guide
Design Immunohistochemistry for CCNE1
Plan CCNE1 staining in paraffin sections using its observed nuclear and cytoplasmic tissue pattern (HPA tissue IHC). This guide covers control tissue selection, consistent fixation, antibody use, chromogenic detection and scoring.
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 and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Variable nuclear and cytoplasmic staining; highest 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 | Low agreement between antibody staining and RNA data (HPA tissue IHC) | |
| Regulation | G1/S cell-cycle role (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended CCNE1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A00543-2) with four published CCNE1 IHC protocols (PMC13604967; PMC5617501; 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 pH 8.0 retrieval (datasheet: A00543-2); the cited studies also report unspecified retrieval or sodium citrate retrieval (PMC13604967; PMC4725452).
Section 2
What Is the Expected CCNE1 Staining Pattern?
CCNE1 is a nuclear cell cycle protein with no transmembrane segment (UniProt P24864). In paraffin section IHC, expect nuclear staining in decidual cells of placenta and cells in testicular seminiferous ducts, where HPA reports high staining; cytoplasmic staining also occurs across tissues (HPA: tissue IHC). Interpret these patterns cautiously: HPA rates its tissue IHC profile Approved but reports low consistency between antibody staining and RNA expression (HPA: reliability).
| Distinct nuclear staining in placental decidual cells or cells in testicular seminiferous ducts. | This matches the strongest listed tissue and cell patterns (HPA: High in both). Assess the named cells, rather than treating every stained cell in the section as an equally informative positive (standard IHC practice). |
| Predominantly cytoplasmic staining with little or no nuclear staining in the expected positive cells. | Review this as a compartment mismatch with the nuclear UniProt assignment and HPA nucleoplasmic IF pattern (UniProt P24864; HPA: subcellular). Cytoplasmic signal alone is not proof of an artifact: HPA also reports variable cytoplasmic tissue staining (HPA: tissue IHC). |
| Strong staining in a cell type listed as not detected, such as liver hepatocytes. | This conflicts with the listed cell specific observation (HPA: hepatocytes Not detected). Check for cross-reactivity, endogenous detection activity, and tissue identity before assigning it to CCNE1 (standard IHC practice). |
| Diffuse color across nuclei, cytoplasm, and tissue spaces, with little cell specific contrast. | This pattern is difficult to score as CCNE1 because the expected nuclear signal lacks clear boundaries (UniProt P24864; standard IHC practice). Review background controls, blocking, antibody concentration, washing, and detection chemistry (standard IHC practice). |
| No staining in placental decidual cells and testicular seminiferous duct cells. | Both are listed as High by HPA, so their absence calls the run into question (HPA: tissue IHC). Confirm that the expected cells are present, then inspect the IHC-validated antibody and detection workflow before calling an experimental sample negative (standard IHC practice). |
💡Expected CCNE1 appearanceCall a positive result when the expected placental decidual or testicular seminiferous duct cells show clear, potentially strong nuclear staining (HPA: High; UniProt P24864); diffuse color or staining confined to an HPA-listed negative cell type is suspect (HPA: tissue IHC; standard IHC practice).
| Compartment | UniProt assigns CCNE1 to the nucleus; HPA tissue IHC reports nuclear and cytoplasmic expression of varying intensity (UniProt P24864; HPA: tissue IHC). Score nuclear and cytoplasmic components separately so a cytoplasmic component does not conceal a weak nuclear result (standard IHC practice). |
| Cell and tissue context | HPA reports High staining in placental decidual cells and testicular seminiferous duct cells, Medium staining in several other listed cell types, and Not detected in others (HPA: tissue IHC). Compare the same cell type across sections; a whole tissue label cannot substitute for cell level assessment (standard IHC practice). |
| Evidence strength | The tissue profile is Approved, with low antibody staining versus RNA consistency; the listed antibodies have IHC Approved status, rather than IHC Enhanced status (HPA: reliability; HPA: antibodies). Treat an isolated unexpected pattern as provisional and check controls and a second validated antibody where available (standard IHC practice). |
| Protein forms | UniProt lists three isoforms and an unprocessed chain spanning residues 1–410, with no signal peptide or propeptide (UniProt P24864). The supplied record does not identify the IHC-validated antibody's epitope or establish whether it detects every isoform; avoid inferring isoform specificity from staining alone. |
| Antigen retrieval | Use retrieval conditions validated for the selected IHC-validated antibody and paraffin section workflow (standard IHC practice). Neither the HPA tissue observations nor the UniProt record establishes CCNE1-specific retrieval requirements or fixation sensitivity (HPA: tissue IHC; UniProt P24864). |
| IF/ICC: where should signal appear? | HPA reports enhanced nucleoplasmic localization and an additional, uncertain cytosolic location in ICC-IF (HPA: subcellular). Use that observation to interpret IF images; this IHC section does not establish an IF/ICC protocol or a cytosolic positive control. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue has no visible signal. | The expected cells may be absent from the section, or an IHC workflow step may have failed (HPA: High in placental decidual and testicular seminiferous duct cells; standard IHC practice). | Locate the named cells on the counterstained section; check the IHC-validated antibody's paraffin section instructions, retrieval, primary incubation, and detection controls (standard IHC practice). |
| A negative cell type appears strongly positive. | Cross-reactivity or endogenous detection activity may mimic staining; compare against the specific HPA cell type rather than its organ alone (HPA: hepatocytes Not detected; standard IHC practice). | Run an appropriate primary-omission control and review detection blocking; seek agreement with another IHC-validated antibody before assigning the signal to CCNE1 (standard IHC practice). |
| Signal appears mainly cytoplasmic. | HPA reports cytoplasmic tissue staining, but nuclear localization is supported by UniProt and HPA ICC-IF (HPA: tissue IHC; UniProt P24864; HPA: subcellular). The cause of cytoplasmic-only staining is unresolved. | Score nuclear and cytoplasmic signal separately, compare a listed High cell type, and review controls for background; avoid calling cytoplasmic staining alone definitive CCNE1 localization (standard IHC practice). |
| The section has diffuse brown background. | Excess nonspecific binding or detection activity can obscure cell boundaries (standard IHC practice); this appearance does not establish a CCNE1 tissue pattern (HPA: tissue IHC). | Inspect the primary-omission control, blocking, antibody concentration, washes, and chromogen development; repeat with conditions validated for the IHC-validated antibody (standard IHC practice). |
| Nuclear staining is weak or uneven across sections. | HPA reports variable intensity across tissues, and its profile has low consistency with RNA expression (HPA: tissue IHC; HPA: reliability). Variation alone does not identify a technical cause. | Compare the same named cell type and a listed High control in the same run; review section quality and staining conditions before interpreting between-section differences (standard IHC practice). |
| Nuclear signal is convincing, but an unexpected cell population also stains. | The extra signal may reflect a real variable tissue pattern or assay background; HPA lists nuclear and cytoplasmic staining in most tissues and provides cell specific levels (HPA: tissue IHC). | Record the unexpected cell type and compartment, compare its HPA entry, and use controls or a second IHC-validated antibody to resolve an interpretation that affects the experiment (standard IHC practice). |
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 as a negative tissue (HPA: Not detected in glandular cells), and compare decidual nuclei with neighboring cells that lack nuclear signal as an internal background reference.
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 a no-primary (secondary-only) control, a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary), and CCNE1 knockout material as a biological negative if available. In placenta, quench endogenous peroxidase and check for endogenous biotin background when using the reported SABC/DAB detection (selected-SKU caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative. The reported placenta IHC used heat-mediated EDTA retrieval at pH 8.0; whether that retrieval is required for other specimens is unreported (selected-SKU caption: EDTA retrieval). ICC-IF images exist (HPA: subcellular), but the supplied evidence does not establish that IF or frozen sections are easier; placenta may show endogenous biotin or peroxidase background with the reported SABC/DAB method (selected-SKU caption: placenta, SABC/DAB).
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 by checking retrieval, controls, and cell-level nuclear localisation before comparing signal across paraffin sections (datasheet A00543-2; UniProt P24864).
How should I retrieve CCNE1 when nuclear staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A00543-2). The selected placenta image then used 10% goat serum block and 1 μg/ml primary antibody overnight at 4°C; use these as reference conditions (datasheet A00543-2). If staining remains weak, compare a citrate-buffer retrieval condition on matched sections as a fallback while keeping antibody and detection conditions fixed (general IHC practice). Examine nuclear signal alongside tissue morphology, using placental decidual cells as a positive reference and a no-primary section to identify detection background (UniProt P24864: nucleus; HPA: high in placental decidual cells; general IHC practice).
Can fixation explain variable CCNE1 staining between paraffin blocks?
The selected paraffin-section caption does not report its fixative, so target-specific CCNE1 fixation sensitivity is unknown (datasheet A00543-2). Record each block’s fixative and fixation duration before comparing staining, since these processing variables can affect antigen accessibility in IHC generally (general IHC practice). Run differently processed blocks through the same EDTA pH 8.0 retrieval and 1 μg/ml antibody condition, with a shared reference section in each batch (datasheet A00543-2; general IHC practice). If signal differs, assess morphology and controls before attributing the difference to CCNE1 abundance; the available caption cannot establish a CCNE1-specific fixation effect (datasheet A00543-2; general IHC practice).
How should I assess CCNE1 signal outside the nucleus?
Prioritise staining in identifiable nuclei because CCNE1 is annotated as nuclear and HPA reports enhanced nucleoplasmic localisation (UniProt P24864; HPA subcellular). HPA also describes cytosolic signal as uncertain and reports nuclear and cytoplasmic tissue staining, so record the two compartments separately (HPA subcellular; HPA tissue IHC). Compare cytoplasmic colour with a no-primary control and inspect whether nuclei retain distinct staining after counterstaining, rather than counting diffuse colour as nuclear positivity (general IHC practice). In placenta, score decidual cells explicitly and use their reported high staining as context, while retaining any reproducible cytoplasmic pattern as a separate observation (HPA tissue IHC: high in placental decidual cells; general IHC practice).
Could CCNE1 isoforms or phosphorylation alter my IHC result?
CCNE1 has 3 annotated isoforms, E1L, E1S, and isoform 3, so the detected forms depend on the antibody epitope (UniProt P24864). The supplied caption gives staining conditions but no epitope map or isoform coverage; avoid assigning a stained cell to one isoform from this IHC image (datasheet A00543-2; UniProt P24864). CCNE1 also has annotated phosphorylation sites, including residues 77, 103, 387, 395, and 399, without evidence here that they change this antibody’s staining (UniProt P24864). When comparing samples, keep retrieval and detection conditions constant and describe the result as CCNE1 immunoreactivity unless epitope and isoform specificity have been established separately (general IHC practice).
How can IF help check the cell and compartment assignments from CCNE1 IHC?
On a separately optimised IF section, multiplex CCNE1 with a validated marker for the expected cell population, such as placental decidual cells, and a nuclear counterstain (HPA tissue IHC: high in placental decidual cells; general IF practice). Choose a spectrally separated red or far-red fluorophore after checking unstained tissue autofluorescence in the intended channels (general IF practice). Because the expected CCNE1 signal is intracellular and mainly nucleoplasmic, optimise permeabilisation to allow antibody access through the cell boundary and into nuclei (UniProt P24864: no transmembrane segment; HPA subcellular: nucleoplasm; general IF practice). Include single-colour and no-primary controls, then compare nuclear and cytosolic signals separately; HPA marks the additional cytosolic location uncertain (HPA subcellular; general IF practice).
What should I check when DAB colour obscures CCNE1-positive nuclei?
First compare the stained section with a no-primary control to separate antibody-associated colour from detection background (general IHC practice). The selected paraffin image used a biotinylated secondary, streptavidin–biotin complex, and DAB, so assess background at each detection step on matched sections (datasheet A00543-2; general IHC practice). Use a peroxidase block and review the blocking step as general chromogenic IHC measures; the caption specifically reports 10% goat serum block but does not report a peroxidase block (datasheet A00543-2; general IHC practice). If colour remains diffuse, review washing, chromogen development, and counterstain before increasing the 1 μg/ml reference antibody concentration (datasheet A00543-2; general IHC practice).
How should I score CCNE1 IHC across samples with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and count evaluable nuclei before scoring, because CCNE1 staining varies across tissues and cell types (HPA tissue IHC; general IHC practice). Report the percentage of positive nuclei and, if intensity matters, a nuclear H-score using intensity categories 0–3, giving a possible range of 0–300 (general IHC practice). Normalise counts to the number of evaluable cells in the same annotated population, or report positive-cell density per mm² of viable annotated tissue (general IHC practice). Keep retrieval, detection, exposure to DAB, and scoring thresholds consistent, and record cytoplasmic staining separately from the nuclear score (datasheet A00543-2; UniProt P24864: nucleus; general IHC practice).
When is apparent CCNE1 positivity likely to be an artefact?
Give greatest weight to reproducible nuclear staining in the expected cells: CCNE1 is nuclear, and HPA reports high staining in placental decidual cells and cells in seminiferous ducts (UniProt P24864; HPA tissue IHC). Treat diffuse colour in an unexpected compartment cautiously, although HPA also reports variable cytoplasmic tissue staining and uncertain cytosolic localisation (HPA tissue IHC; HPA subcellular). Check section edges, necrotic areas, and no-primary controls for staining patterns caused by processing or endogenous detection activity (general IHC practice). HPA rates tissue staining Approved but notes low consistency with RNA data, so corroborate a surprising biological conclusion with controls and an independent measure (HPA tissue IHC; general IHC practice).
Boster reagents
Best CCNE1 / G1/S-specific cyclin-E1 IHC Antibodies
These anti-CCNE1 antibodies have IHC images from human placenta and rat colon sections, plus an IF image from human HeLa cells (catalog image captions).
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 has IHC data from paraffin sections of human placenta, human lung cancer, and mouse testis (A00543-2 image captions). M00543-4 has IHC images from rat and human colon, while M00543 has an IF image from HeLa cells (M00543-4 and M00543 image captions).
Which to pick: For paraffin-section IHC, choose A00543-2 for its documented EDTA retrieval at pH 8.0 and 1 μg/ml primary incubation; the caption does not report the fixative (A00543-2 IHC caption). For IF/ICC, choose rabbit monoclonal M00543, which lists both applications and has a HeLa-cell IF image (M00543 catalog entry and image caption). For an antibody listed for both tissue IHC and IF across human, mouse, and rat, choose rabbit monoclonal M00543-4; its rat-colon IHC caption does not report section processing or fixative (M00543-4 catalog entry and IHC caption).
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 and cytoplasmic tissue staining (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: Low agreement between antibody staining and RNA data (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: G1/S cell-cycle role (UniProt). Evidence: UniProt P24864, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A00543-2, PMC13604967, PMC5617501. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCNE1/immunohistochemistry-ihc-guide.