CDC27 / Cell division cycle protein 27 homolog · IHC design guide

Design Immunohistochemistry for CDC27

Plan chromogenic CDC27 IHC in paraffin sections using nuclear staining as the expected tissue pattern (HPA tissue IHC). Compare high-staining hematopoietic cells with low-staining hepatocytes (HPA tissue IHC), and start the catalog antibody at 2–5 μg/mL (datasheet A03905-3).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for CDC27 (IHC for CDC27): expected localisation Nuclear staining in tissue (HPA tissue IHC), antibody A03905-3, validated IHC image, and IHC protocol steps — Printable **CDC27** IHC protocol sheet — expected localisation Nuclear staining in tissue (HPA tissue IHC), antibody A03905-3, controls and protocol steps. Open the full CDC27 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

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	Nuclear staining in tissue (HPA tissue IHC)
	Staining pattern	Widespread 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 (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03905-3)
	Caveat	Staining intensity varies by cell type (HPA tissue IHC)
	Regulation	No expression regulator annotated (UniProt)
	Isoform / epitope	2 isoforms; epitope coverage is unknown (UniProt)

Section 1

Recommended CDC27 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by four published CDC27 IHC workflows, with each article’s reported steps listed below (datasheet A03905-3; PMC9185143; PMC4955705; PMC5351633; PMC8886130).

Recommended immunohistochemistry (IHC-P) protocol parameters
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 pH 8.0 retrieval (datasheet A03905-3). Citrate pH 6.0 is a published alternative (PMC9185143; PMC8886130).

Section 2

What Is the Expected CDC27 Staining Pattern?

CDC27 should show predominantly nuclear staining across many cell types in paraffin-section IHC (HPA: ubiquitous nuclear expression; reliability Supported). High staining is reported in selected hematopoietic, epithelial, glandular, endothelial and kidney tubular cells (HPA: tissue IHC). A spindle-associated cytoplasmic pool is also annotated (UniProt P30260: nucleus; cytoplasm, cytoskeleton, spindle). CDC27 has no transmembrane segment, so a membrane-rim pattern is not expected (UniProt P30260: topology).

What am I looking at on my slide?
Distinct nuclear chromogen in the expected cells, with low background.	This fits the reported IHC profile; compare cell types within the section rather than requiring equal intensity throughout (HPA: ubiquitous nuclear expression; variable tissue IHC levels).
Predominant membrane staining, or cytoplasmic staining without a convincing nuclear pattern.	A membrane rim conflicts with the reported localisation (UniProt P30260: no transmembrane segment; HPA: nuclear IHC). Investigate nonspecific staining. Assess possible spindle staining only in mitotic cells (UniProt P30260: spindle).
Strong staining in cells expected to have little or no CDC27 signal, especially cardiomyocytes.	HPA reports CDC27 as not detected in cardiomyocytes (HPA: heart muscle IHC). Check cell identity and controls; cross-reactivity or endogenous detection activity are possible explanations (general IHC practice).
Diffuse chromogen over nuclei, cytoplasm and surrounding tissue.	This cannot establish the expected nuclear pattern (HPA: tissue IHC). Excess antibody, incomplete blocking or endogenous detection activity can cause background (general IHC practice).
No nuclear signal in a known-positive cell population.	This conflicts with its reported high staining (HPA: bone-marrow hematopoietic cells; kidney tubular cells). First verify tissue identity and positive-control performance, then review the IHC workflow (general IHC practice).

💡Expected CDC27 appearanceCall positive when expected cells show distinct nuclear staining, potentially high in HPA-listed high-staining populations; a dominant membrane rim or staining confined to cardiomyocytes is suspect (HPA: tissue IHC; UniProt P30260: topology).

How each factor affects the staining
Tissue and cell selection	High IHC staining is reported in bone-marrow hematopoietic cells, bronchial basal cells, colon endothelial cells and kidney tubular cells; hepatocytes, prostate glandular cells and ovarian follicle cells are low (HPA: tissue IHC).
Cellular compartment	Ubiquitous nuclear expression is the tissue-IHC reference (HPA: tissue IHC). UniProt also annotates cytoplasm and spindle, so evaluate their staining in cellular context rather than treating every extranuclear pixel as specific (UniProt P30260: localisation).
Antibody evidence	HPA reports IHC staining supported for HPA028129 and CAB016315; the supplied record gives no IHC status for HPA052399 or HPA065835 (HPA: antibody validation). Validation for one antibody does not establish another antibody’s IHC performance (general IHC practice).
Isoforms and epitope	Two isoforms are listed, but the supplied record gives no antibody epitope or isoform-specific staining result (UniProt P30260: isoforms 1 and 2). Check the chosen antibody’s documented epitope before making an isoform-specific interpretation (general IHC practice).
Processing and retrieval	The record lists no signal peptide, propeptide or cleavage into separate chains (UniProt P30260: processing). Antigen retrieval is a general paraffin-IHC workflow variable; no CDC27-specific fixation sensitivity or retrieval condition is established here (general IHC practice).
IF/ICC Q&A: Where should CDC27 appear?	Predominantly in the nucleoplasm in the separate IF/ICC application (HPA: nucleoplasm, enhanced). HPA lists ICC-IF images in A-431, U-251MG, U2OS, HEL and Rh30; those observations do not supply an IHC-P protocol (HPA: subcellular record).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Nuclear signal is absent in a reported high-staining population.	The section, detection run or cell identification may be at fault (general IHC practice); absence conflicts with HPA’s high IHC observation.	Confirm the relevant cells are present, inspect a positive control and review the catalog antibody’s IHC-P conditions (HPA: high-staining populations; general IHC practice).
Chromogen outlines cell membranes.	A dominant membrane pattern is inconsistent with CDC27’s annotated localisation and lack of a transmembrane segment (UniProt P30260: localisation and topology).	Compare against a no-primary control and reassess specificity and detection conditions (general IHC practice).
Cardiomyocytes stain strongly.	HPA reports cardiomyocytes as not detected; cross-reactivity or endogenous detection activity may contribute (HPA: heart muscle IHC; general IHC practice).	Verify cell identity and inspect no-primary and detection controls before assigning the signal to CDC27 (general IHC practice).
Diffuse brown background obscures nuclei.	Background can reflect antibody concentration, blocking or endogenous detection activity (general IHC practice).	Review the catalog antibody’s IHC-P dilution and detection steps, then compare with a no-primary control (general IHC practice).
Only low-staining tissue appears negative.	A weak result may match the reference: hepatocytes, prostate glandular cells and ovarian follicle cells are low (HPA: tissue IHC).	Assess a documented high-staining cell population alongside it before concluding the run failed (HPA: tissue IHC; general IHC practice).
An IF/ICC image seems different from the IHC slide.	The IF/ICC reference is nucleoplasmic, while tissue IHC is described as ubiquitous nuclear expression; the methods show different preparations (HPA: subcellular; tissue IHC).	Interpret each against its own HPA application record and use the catalog antibody’s IHC-P workflow for the paraffin section (HPA: antibody validation; general IHC practice).

Source: UniProt P30260 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CDC27 staining

IHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human bladder cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CDC27 Antibody (A03905-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A03905-3 · IHCIHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human bladder cancer tissue.

IHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human gastric cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CDC27 Antibody (A03905-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

A03905-3 · IHCIHC analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in paraffin-embedded section of human gastric cancer tissue.

IF analysis of CDC27 using anti-CDC27 antibody (A03905-3). CDC27 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-CDC27 Antibody (A03905-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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.

← 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 score hematopoietic cells for CDC27 staining (HPA: High in bone marrow hematopoietic cells). Use heart muscle cardiomyocytes as the tissue negative (HPA: Not detected); on the bone marrow slide, cells without specific signal should show only background staining, without assigning an unsupported cell type as an internal negative.

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 a no-primary, secondary-only control and a concentration-matched nonimmune rabbit IgG control (caption: rabbit anti-CDC27 primary antibody). Use a verified CDC27 knockout sample or a validated peptide-block control as a biological negative; with the caption’s biotin-based DAB detection, check endogenous peroxidase and biotin background in bone marrow (caption: biotinylated secondary, SABC and DAB).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03905-3 caption does not state the fixative (caption: paraffin-embedded section; fixative unreported). The reported IHC procedure uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish that retrieval is required for every preparation (caption: heat-mediated EDTA retrieval). No supplied evidence shows that frozen sections or IF are easier; HPA ICC-IF images show nucleoplasmic localization, while endogenous peroxidase or biotin may complicate interpretation of the caption’s biotin-based DAB staining in bone marrow (HPA: Nucleoplasm; caption: SABC and DAB; standard IHC practice).

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

Use compartment, tissue controls, and the documented paraffin-section workflow to troubleshoot CDC27 staining in chromogenic IHC.

What should I change if CDC27 staining is weak after antigen retrieval?

Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (A03905-3 image caption). The documented section was then incubated with 2 μg/mL antibody overnight at 4°C, so check retrieval consistency and incubation conditions before increasing antibody concentration (A03905-3 image caption; standard IHC practice). Include a known positive section in each run and compare nuclear signal with background, because CDC27 shows ubiquitous nuclear tissue staining and enhanced nucleoplasmic localization (HPA tissue IHC; HPA subcellular). If staining remains weak, test a different retrieval buffer on matched sections as a fallback, changing one condition at a time and checking tissue preservation (standard IHC practice).

Could fixation be responsible for weak CDC27 staining?

The selected paraffin-section image does not state its fixative, so CDC27-specific sensitivity to fixation is unknown (A03905-3 image caption). Record the fixative, fixation duration, processing schedule, and section age for each specimen before comparing staining runs (standard IHC practice). Evaluate suspect and reference sections together using the documented EDTA pH 8.0 retrieval and 2 μg/mL overnight antibody incubation, while keeping detection conditions constant (A03905-3 image caption; standard IHC practice). If signal differs, use matched samples with controlled fixation histories to investigate the cause; the available image cannot establish that fixation caused the difference (A03905-3 image caption; standard IHC practice).

How should I assess nuclear and cytoplasmic CDC27 staining?

Score nuclear staining first: tissue IHC reports ubiquitous nuclear expression, and cell imaging places CDC27 predominantly in the nucleoplasm (HPA tissue IHC; HPA subcellular). UniProt also lists cytoplasm, cytoskeleton, and spindle, so a cytoplasmic or mitotic pattern merits examination in its cellular context rather than automatic rejection (UniProt P30260 subcellular). Use a hematoxylin counterstain and inspect intact cell borders and mitotic figures at higher magnification before assigning a compartment (standard IHC practice). Record nuclear and cytoplasmic scores separately, and compare unusual patterns with positive tissue and secondary-only controls processed in the same run (HPA tissue IHC; standard IHC practice).

Can isoforms or phosphorylation explain inconsistent CDC27 staining?

CDC27 has 2 annotated isoforms and multiple modified residues, including phosphorylation at positions 205, 209, and 244 (UniProt P30260 isoforms; UniProt P30260 modified residues). The supplied image does not identify the antibody epitope or establish isoform-specific detection, so do not assign a staining difference to either mechanism from IHC alone (A03905-3 image caption). Check the antibody's documented immunogen or epitope, if available, against isoform sequences before interpreting discordant specimens (standard antibody-validation practice). Compare matched sections across retrieval conditions and use an independently validated reagent or orthogonal assay when isoform or modification specificity matters (standard IHC practice).

How can I follow up an IHC pattern with multiplex IF?

For follow-up IF, pair CDC27 with a validated marker for the cell population under study and compare signal within individual cells; tissue IHC and cell imaging support a predominantly nuclear pattern (HPA tissue IHC; HPA subcellular; standard IF practice). Choose fluorophores after inspecting unstained tissue in each channel, and place the weaker signal in a channel with less autofluorescence (standard IF practice). CDC27 has no annotated transmembrane segment, so use permeabilisation appropriate for access to its intracellular, especially nucleoplasmic, epitopes (UniProt P30260 topology; HPA subcellular; standard IF practice). Titrate permeabilisation and antibodies on controls, and check single-stain controls for bleed-through before interpreting colocalisation (standard IF practice).

What causes diffuse or patchy background in CDC27 chromogenic IHC?

First compare the section with a secondary-only control and inspect whether DAB deposits follow tissue edges, damaged areas, or endogenous pigment rather than intact nuclei (standard IHC practice; HPA tissue IHC). The documented workflow used 10% goat serum blocking, a biotinylated secondary, a streptavidin-biotin complex, and DAB, so examine blocking and detection steps when background rises (A03905-3 image caption). Include an appropriate endogenous peroxidase block and assess endogenous biotin where relevant to that detection system (standard IHC practice). Adjust washing, antibody concentration, and chromogen development one variable at a time, judging each change against preserved nuclear signal in a positive section (standard IHC practice; HPA tissue IHC).

How should I quantify CDC27 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the tissue compartment and cell population before scoring, because CDC27 tissue IHC shows nuclear expression across many tissues but differing levels among cell types (HPA tissue IHC). For each annotated region, report the percentage of positive nuclei and, when intensity is reproducible, an H-score using the same intensity thresholds across slides (standard IHC quantification practice). Normalize positive-cell counts to the number of eligible intact nuclei, or report positive-cell density per mm² of viable tissue when cell density is itself relevant (standard IHC quantification practice). Exclude folds and necrosis consistently, and keep acquisition, counterstain, and scoring rules fixed across comparison groups (standard IHC quantification practice).

When is an unexpected CDC27-positive area likely to be artefactual?

Give greatest weight to staining in intact nuclei: HPA reports ubiquitous nuclear tissue expression and enhanced nucleoplasmic localization, while UniProt also allows cytoskeletal and spindle localization (HPA tissue IHC; HPA subcellular; UniProt P30260 subcellular). Check cell identity before calling an area aberrant, since reported high staining includes kidney tubular cells and colon endothelial cells, whereas cardiomyocytes were not detected (HPA tissue IHC). Treat staining confined to section edges, necrosis, or the secondary-only control as suspect, and compare it with intact internal cells (standard IHC practice). If diffuse DAB obscures nuclei, investigate endogenous peroxidase and detection background before assigning CDC27 positivity (standard IHC practice).

Boster reagents

Best CDC27 / Cell division cycle protein 27 homolog IHC Antibodies

The catalog includes CDC27 tissue IHC images from human paraffin sections and rat liver (catalog IHC captions), plus an IF image from human A431 cells (A03905-3 IF caption).

Real IHC data

Anti-CDC27 Antibody ®

Cat # A03905-3

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Real IHC data Immunohistochemistry of APC3 in rat liver tissue with APC3 antibody at 5 μg/mL.

Anti-APC3 CDC27 Antibody

Cat # A03905

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A03905-3 will render with human bladder cancer paraffin-section IHC; its separate IF image uses A431 cells (A03905-3 image captions). A03905 will render with rat liver IHC and lists IHC-P as an application (A03905 image caption; catalog applications).

Which to pick: Choose A03905-3 for human paraffin-section IHC: its image caption documents EDTA retrieval and 2 μg/mL primary antibody, but does not report the fixative (A03905-3 IHC caption). Choose A03905-3 for IF/ICC because both applications and an A431 cell IF image are listed; A03905 has no IF application or image (catalog applications; A03905-3 IF caption). For work across species, both list human, mouse and rat reactivity, while the shown tissue IHC evidence covers human for A03905-3 and rat for A03905; clonality is unreported for both (catalog reactivity and clone fields; IHC 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.