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CDC27 / Cell division cycle protein 27 homolog · IHC design guide
Design Immunohistochemistry for CDC27
For paraffin CDC27 IHC, start with 2–5 μg/mL A03905-3 (datasheet: IHC-P). Expect widespread nuclear staining, with high signal in bone marrow hematopoietic cells and no detected signal in cardiomyocytes (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CDC27 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Ubiquitous nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03905-3) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Heart muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Cardiomyocytes may lack detectable staining (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 2 isoforms; effect on IHC epitope recognition unknown (UniProt) |
Section 1
Recommended CDC27 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A03905-3) is accompanied by four published CDC27 IHC workflows (PMC9185143; PMC4955705; PMC5351633; PMC8886130).
| Sample | Paraffin-embedded human bladder cancer tissue; fixative not specified (datasheet A03905-3) |
| Fixation | Image fixative and duration unreported (datasheet A03905-3); 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 A03905-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03905-3) |
| Primary antibody | Rabbit anti-CDC27, 2-5μg/ml (datasheet A03905-3) |
| Primary incubation | Overnight at 4 °C (datasheet A03905-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03905-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDC27-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A03905-3); the published protocols use different retrieval conditions.
Section 2
What Is the Expected CDC27 Staining Pattern?
Expect CDC27 staining chiefly in nuclei across many tissue cell types: HPA describes ubiquitous nuclear expression and rates its tissue IHC profile Supported because staining agrees with RNA data (HPA: tissue IHC). High staining is reported in selected hematopoietic, epithelial, glandular, endothelial, and tubular cells (HPA: tissue IHC). UniProt also places CDC27 in the cytoplasm and spindle and reports no transmembrane segment (UniProt P30260: subcellular location and topology).
| Clear nuclear chromogen in kidney tubular cells or bone marrow hematopoietic cells, with cell boundaries still discernible. | This matches the expected compartment and two reported high-staining populations (HPA: ubiquitous nuclear expression; High in kidney tubular and bone marrow hematopoietic cells). Compare the intended cells with adjacent structures when reading a section; a positive tissue control supports interpretation of the run (general IHC practice). |
| Predominantly cytoplasmic staining across a section, with little nuclear signal. | This departs from the reported tissue IHC profile (HPA: ubiquitous nuclear expression). Check the staining controls and scoring before calling it CDC27. UniProt also lists cytoplasmic and spindle locations, so a localized nonnuclear signal alone cannot establish an artefact; the supplied evidence does not define its expected frequency in tissue sections (UniProt P30260: subcellular location). |
| Strong signal in heart muscle cardiomyocytes, while the expected nuclear pattern is absent. | HPA reports CDC27 as not detected in cardiomyocytes; assess possible cross-reactivity or endogenous detection activity with appropriate controls (HPA: heart muscle cardiomyocytes, Not detected; general IHC practice). That observation concerns cardiomyocytes specifically and should not be extended to every cell in a heart section (HPA: tissue IHC). |
| Chromogen appears broadly across nuclei, cytoplasm, and tissue spaces, obscuring individual cells. | This cannot be scored confidently as the HPA nuclear pattern (HPA: ubiquitous nuclear expression). Assess background with a control omitting the primary antibody, then review blocking, washing, and detection conditions; these are general IHC checks, not established CDC27-specific effects (general IHC practice). |
| No nuclear signal in kidney tubular cells or bone marrow hematopoietic cells. | These populations are reported High and are useful positive tissue checks (HPA: kidney tubular and bone marrow hematopoietic cells, High). A failed signal warrants review of the antibody, retrieval, and detection run before inferring absence of CDC27; the supplied sources do not establish a CDC27-specific retrieval requirement (general IHC practice). |
💡Expected CDC27 appearanceCall a result positive when distinct nuclear staining is visible in an expected cell population, especially one rated High by HPA, such as kidney tubular cells; widespread cytoplasmic-only signal or staining that obscures cells warrants control review before scoring (HPA: ubiquitous nuclear expression; kidney tubular cells, High; general IHC practice).
| Cell population chosen for interpretation (HPA: tissue IHC). | HPA reports High staining in bronchial basal cells, colon endothelial cells, and several other listed populations; hepatocytes, prostate glandular cells, and ovarian follicle cells are Low, while cardiomyocytes are Not detected (HPA: tissue IHC). Score the named population rather than treating a whole organ as uniformly positive or negative (general IHC practice). |
| Antibody validation (HPA: antibody records). | HPA rates tissue IHC for HPA028129 and CAB016315 as Supported; its ICC ratings are Enhanced and Supported, respectively (HPA: antibody records). These are application-specific validation summaries, so an ICC rating does not establish how an antibody will stain a paraffin section (HPA: antibody records; general IHC practice). |
| Epitope and CDC27 variants (UniProt P30260: isoforms and modified residues). | UniProt lists 2 isoforms and 15 modified residues, including phosphorylation sites (UniProt P30260: isoforms and modified residues). No antibody epitope or variant-specific staining result is supplied, so these features cannot predict a particular IHC intensity or establish why a tissue is negative. |
| Processing and cellular topology (UniProt P30260: processing and topology). | UniProt lists a single chain spanning residues 1–824, with no signal peptide, propeptide, or transmembrane segment (UniProt P30260: processing and topology). Those annotations support interpreting CDC27 as an intracellular target; they do not establish an antigen-retrieval setting or a tissue-specific staining level. |
| IF/ICC Q&A: Where should signal appear? | In the supplied ICC-IF profile, HPA identifies the nucleoplasm as the enhanced main location; UniProt additionally lists cytoplasm and spindle (HPA: subcellular ICC-IF; UniProt P30260: subcellular location). This is an IF/ICC localisation answer, not a paraffin IHC protocol or a reason to expect every compartment in every cell. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known high-staining population is blank. | The run may have failed, or the chosen antibody and conditions may not produce detectable signal in that section; these are possibilities, not CDC27-specific findings (general IHC practice). | Check a positive tissue control and the detection reagents; review the antibody instructions and antigen-retrieval conditions used for paraffin IHC (general IHC practice). Kidney tubular cells are reported High, but no CDC27-specific retrieval condition is supplied (HPA: kidney tubular cells, High). |
| Most cells show cytoplasmic staining without convincing nuclei. | The pattern conflicts with HPA's ubiquitous nuclear tissue profile, although UniProt also records cytoplasmic and spindle localisation (HPA: tissue IHC; UniProt P30260: subcellular location). | Review the positive control and compare nuclear detail with the counterstain; assess background and antibody specificity before assigning the signal to CDC27 (general IHC practice). |
| Cardiomyocytes stain strongly. | That result disagrees with HPA's Not detected call for cardiomyocytes; cross-reactivity or endogenous detection activity is possible, rather than established by this observation alone (HPA: heart muscle cardiomyocytes, Not detected; general IHC practice). | Identify the stained cell type carefully and run an appropriate control omitting the primary antibody; inspect detection-related background before scoring (general IHC practice). |
| Staining is diffuse and cell boundaries are hard to read. | Background may be obscuring the nuclear pattern reported by HPA (HPA: ubiquitous nuclear expression; general IHC practice). | Compare a control omitting the primary antibody; review blocking, washing, and detection conditions, then score only signal whose cellular location can be resolved (general IHC practice). |
| A low-staining tissue appears weaker than the positive control. | This can agree with HPA: hepatocytes, prostate glandular cells, and ovarian follicle cells are Low, whereas kidney tubular cells are High (HPA: tissue IHC). | Compare the named cell populations under the same run conditions and report intensity with its cell type; weak staining alone does not prove technical failure (HPA: tissue IHC; general IHC practice). |
| An IF/ICC image shows nucleoplasmic signal, but the IHC section looks different. | HPA's enhanced nucleoplasmic location comes from ICC-IF, while its tissue IHC summary reports ubiquitous nuclear expression; the records describe different applications (HPA: subcellular ICC-IF and tissue IHC). | Interpret the paraffin section against the tissue IHC profile and its controls. Use the ICC-IF location as supporting context, without treating it as an IHC protocol or an exact intensity standard (HPA: tissue IHC and subcellular ICC-IF; general IHC practice). |
Experiment evidence
IHC & IF images showing CDC27 staining
A03905-3 · IHCIHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human bladder cancer tissue.
A03905-3 · IHCIHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human gastric cancer tissue.
A03905-3 · IFIF analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in immunocytochemical section of A431 cells. Enzyme antigen re
A03905 · IHCImmunohistochemistry of APC3 in rat liver tissue with APC3 antibody at 5 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000004897-CDC27/tissue/cerebral+cortex#imid_4019444
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000004897-CDC27/tissue/esophagus#imid_4019408
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000004897-CDC27/tissue/colon#imid_4010614
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000004897-CDC27/tissue/liver#imid_4010615
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB016315; green = CDC27, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB016315; green = CDC27, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CDC27 IHC & IF
🧪Run bone marrow first and require staining in hematopoietic cells (High; HPA: bone marrow). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes). On the bone marrow slide, cells without specific staining should show only background; the supplied HPA rows do not identify a particular internal cell type as CDC27-negative (HPA: bone marrow).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CDC27 in A-431, U-251MG, U2OS, HEL, Rh30, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), host-matched rabbit IgG isotype, and CDC27-knockout material as controls (caption: rabbit primary antibody; standard IHC practice). Quench endogenous peroxidase and assess endogenous biotin in bone marrow, especially with the caption’s biotin-based DAB detection (HPA: bone marrow hematopoietic cells; caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03905-3 paraffin-section caption does not state the fixative (caption: fixative unreported). The demonstrated IHC starting condition uses heat-mediated EDTA pH 8.0 retrieval and 2 µg/mL primary antibody overnight at 4°C; the evidence does not establish whether retrieval is essential or whether frozen sections or IF are easier (caption: retrieval and incubation; HPA: nucleoplasmic ICC-IF signal). Bone marrow may require particular attention to endogenous peroxidase and biotin background with this detection system (HPA: hematopoietic cells; caption: SABC/DAB).
HPA tissue IHC evidence for CDC27
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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDC27 IHC Tips
Troubleshoot CDC27 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
How should I retrieve CDC27 in paraffin sections when nuclear staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03905-3). The selected CDC27 tissue image used this retrieval before incubation with 2 μg/ml primary antibody overnight at 4°C, so use it as the initial benchmark for this antibody (datasheet A03905-3). If staining remains weak, compare a small retrieval-time series on adjacent sections while holding antibody concentration and detection constant; excessive heating can damage morphology (standard IHC practice). Judge improvement by crisp nuclear signal in viable cells and stable background, using the same positive-control section in each run (HPA: ubiquitous nuclear expression; standard IHC practice).
Can I attribute weak CDC27 staining to the tissue fixative?
Target-specific CDC27 sensitivity to fixation is unknown from the supplied evidence, and the selected paraffin-section caption does not report a fixative (datasheet A03905-3). Record each specimen’s fixative and fixation duration, then compare sections processed together before assigning weak signal to fixation (standard IHC practice). Keep retrieval at EDTA pH 8.0 and start with the documented 2 μg/ml primary antibody condition so that a processing comparison has a consistent baseline (datasheet A03905-3). Include a known staining control on the same run, and assess nuclear signal alongside preserved morphology; a failed control calls for technical troubleshooting before biological interpretation (HPA: ubiquitous nuclear expression; standard IHC practice).
How should I evaluate cytoplasmic CDC27 staining when the expected signal is nuclear?
Prioritise distinct nuclear staining when reading chromogenic CDC27 sections: tissue IHC shows ubiquitous nuclear expression, and subcellular imaging places CDC27 in the nucleoplasm (HPA tissue IHC; HPA subcellular). Cytoplasmic signal is biologically possible because CDC27 is also annotated in the cytoplasm and spindle, so record it separately from nuclear signal (UniProt P30260 localisation). Check whether cytoplasmic colour follows cell boundaries in intact tissue or instead pools in damaged areas, and compare it with a no-primary control (standard IHC practice). If cytoplasmic staining dominates while the control lacks expected nuclear staining, review retrieval, antibody concentration and detection background before assigning that pattern to CDC27 (HPA tissue IHC; standard IHC practice).
Can this IHC stain distinguish CDC27 isoforms or phosphorylation states?
Do not assign an isoform or phosphorylation state from this stain without epitope-specific validation: CDC27 has 2 annotated isoforms and multiple modified residues (UniProt P30260 isoforms and modified residues). The supplied tissue caption documents staining conditions but gives no epitope mapping or isoform selectivity for A03905-3 (datasheet A03905-3). CDC27 has no annotated transmembrane segment, so membrane-only colour would also need careful technical review rather than an epitope-based explanation (UniProt P30260 topology; standard IHC practice). For a specific isoform or phosphosite claim, establish which sequence the antibody recognises and validate discrimination with appropriate controls before interpreting chromogenic intensity (standard IHC practice).
How can I check a CDC27 IHC pattern with multiplex immunofluorescence?
Use IF as a separate validation experiment and compare CDC27 with a marker chosen for the cell population being evaluated; HPA reports nuclear CDC27 across tissues, including high staining in bronchial basal cells (HPA tissue IHC). Place the CDC27 fluorophore in a channel with low tissue autofluorescence and inspect unstained tissue plus single-colour controls before judging colocalisation (standard IF practice). CDC27 is annotated in the nucleus, cytoplasm and spindle, with no transmembrane segment, so optimise permeabilisation for access to intracellular epitopes after checking the antibody’s IF suitability (UniProt P30260 localisation and topology; standard IF practice). Score nuclear overlap within identified cells, and treat the IHC caption’s EDTA pH 8.0 retrieval as evidence for that paraffin-section workflow only (datasheet A03905-3; standard IF practice).
What should I change when CDC27 DAB staining is diffuse or uneven?
First compare the section with a no-primary control and inspect whether diffuse DAB persists, especially around tissue edges or damaged regions (standard IHC practice). The documented chromogenic workflow used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a biotinylated secondary, and streptavidin-biotin detection with DAB (datasheet A03905-3). Check peroxidase blocking, washing and detection reagent exposure as general IHC controls; excess signal from these steps can obscure cellular localisation (standard IHC practice). Reduce primary concentration or development time in a controlled comparison only after the positive control remains detectable, and favour discrete nuclear staining over uniform haze (HPA tissue IHC; standard IHC practice).
How should I score CDC27 staining across sections with different cellularity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and viable tissue area before scoring, then report the percentage of positive nuclei and nuclear intensity using a consistent threshold (HPA tissue IHC: ubiquitous nuclear expression; standard IHC practice). An H-score combines percentages at intensity grades 0–3 into a 0–300 score; state the grades and whether cytoplasmic staining was excluded (standard IHC practice). Normalise positive-cell counts to the number of eligible cells, or report density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Keep retrieval, antibody concentration, DAB development and imaging settings consistent across cases, and use control sections to flag batch effects (standard IHC practice).
What evidence separates a true CDC27 positive cell from staining artefact?
A convincing positive cell has interpretable nuclear staining in intact tissue, consistent with ubiquitous nuclear tissue expression and enhanced nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Compare the stained cell type with its surrounding morphology: HPA reports high signal in several cell populations, including colon endothelial cells, but cardiomyocytes were not detected in its tissue panel (HPA tissue IHC). Treat colour confined to edges, necrotic areas or a no-primary control as suspect, and check peroxidase blocking when DAB appears without specific primary staining (standard IHC practice). CDC27 can also occupy the cytoplasm and spindle, so a nonnuclear pattern requires separate validation rather than automatic rejection (UniProt P30260 localisation; standard IHC practice).
Boster reagents
Best CDC27 / Cell division cycle protein 27 homolog IHC Antibodies
The catalog shows CDC27 IHC data from human paraffin sections and rat liver, plus IF/ICC data from A431 cells (catalog image captions).
Real IHC data
Anti-CDC27 Antibody ®
Cat # A03905-3
Real IHC data
Anti-APC3 CDC27 Antibody
Cat # A03905
A03905 has a rat liver IHC image at 5 μg/mL (A03905 IHC image caption). A03905-3 has human bladder cancer paraffin-section IHC data and A431-cell IF/ICC data (A03905-3 image captions).
Which to pick: For human tissue IHC, choose A03905-3: its image shows a paraffin-embedded bladder cancer section, with the fixative unreported (A03905-3 IHC image caption). For rat tissue IHC, A03905 has a rat liver image; for IF/ICC, choose A03905-3, which lists those applications and has an A431-cell IF image (A03905 IHC image caption; A03905-3 application list and IF image caption). For cross-species planning, both list Human, Mouse, and Rat reactivity, while the shown IHC images document human tissue for A03905-3 and rat tissue for A03905; both are rabbit antibodies with no clone reported (catalog reactivity, host and clone fields; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30260 (CDC27_HUMAN, Cell division cycle protein 27 homolog).
- Human Protein Atlas. CDC27 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CDC27 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. CDC27 antibody validation summary (4 antibodies).
- Prognostic and clinical significance of subcellular CDC27 for patients with rectal adenocarcinoma treated with adjuvant chemotherapy. Oncology letters 2022 — PMC9185143.
- PCBP1/HNRNP E1 Protects Chromosomal Integrity by Translational Regulation of CDC27. Molecular cancer research : MCR 2016 — PMC4955705.
- mir-218-2 promotes glioblastomas growth, invasion and drug resistance by targeting CDC27. Oncotarget 2017 — PMC5351633.
- CDC27-ODC1 Axis Promotes Metastasis, Accelerates Ferroptosis and Predicts Poor Prognosis in Neuroblastoma. Frontiers in oncology 2022 — PMC8886130.
- PubMed PMID:8234252 — UniProt-cited evidence.
- PubMed PMID:7756179 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CDC27 (Cell division cycle protein 27 homolog). Expected localisation Predominantly nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03905-3). Validated IHC antibody: A03905-3 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human bladder cancer tissue; fixative not specified (datasheet A03905-3); 2. Fixation: Image fixative and duration unreported (datasheet A03905-3); 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 A03905-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03905-3); 8. Primary antibody: Rabbit anti-CDC27, 2-5μg/ml (datasheet A03905-3); 9. Primary incubation: Overnight at 4 °C (datasheet A03905-3). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Cardiomyocytes may lack detectable staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; effect on IHC epitope recognition unknown (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P30260, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet A03905-3, PMC9185143, PMC4955705. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CDC27/immunohistochemistry-ihc-guide.