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CDCA5 / Sororin · IHC design guide
Design Immunohistochemistry for CDCA5
Plan paraffin-section CDCA5 IHC using high-staining bone marrow hematopoietic cells as a positive control (HPA tissue IHC). Interpret predominantly nuclear staining and occasional cytoplasmic signal alongside CDCA5’s cell-cycle-dependent chromatin association (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CDCA5 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 | Mostly nuclear; cytoplasmic in some tissues (HPA tissue IHC) | |
| Staining pattern | Germinal center cells: nuclear; some tissues: cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Nuclear envelope breakdown can shift CDCA5 into cytoplasm (UniProt) | |
| Regulation | S phase–metaphase chromatin binding (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–252 chain (UniProt) |
Section 1
Recommended CDCA5 IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol with four published CDCA5 IHC protocols (PMC6267780; PMC9688237; PMC4823039; PMC11214526).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet M05043) |
| Fixation | Image fixative and duration unreported (datasheet M05043); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit monoclonal (clone IAH-3) anti-CDCA5, 1:50 (datasheet M05043) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CDCA5-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in a majority of tissues, including testis and lymphoid tissues. Cytoplasmic expression in a few tissues e.g. placenta. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page antigen retrieval); the cited breast and oral cancer protocols used citrate pH 6.0 (PMC9688237; PMC4823039).
Section 2
What Is the Expected CDCA5 Staining Pattern?
CDCA5 is mainly nuclear in tissue IHC, with cytoplasmic staining reported in a few tissues, including placenta (HPA tissue IHC). Expect staining in the cell populations HPA identifies, such as lymph-node germinal-center cells and bone-marrow hematopoietic cells (HPA tissue IHC). CDCA5 associates with chromatin during the cell cycle and has no transmembrane segment (UniProt Q96FF9). HPA rates the tissue pattern Enhanced, with medium consistency against RNA data (HPA tissue IHC).
| Distinct nuclear staining in germinal-center cells or hematopoietic cells, with other cells less prominent (HPA tissue IHC). | This fits the reported compartment and high-staining cell populations (HPA tissue IHC). Judge the result by cell identity and location as well as intensity; HPA reports nuclear expression across a majority of tissues (HPA tissue IHC). |
| Predominantly membrane staining, or widespread cytoplasmic staining outside a reported context (UniProt Q96FF9; HPA tissue IHC). | Membrane localisation is unsupported by the no-transmembrane topology (UniProt Q96FF9). Investigate cytoplasmic signal in context: HPA reports it in a few tissues, while UniProt describes cytoplasmic release after nuclear-envelope breakdown (HPA tissue IHC; UniProt Q96FF9). |
| Strong staining in adipocytes, cardiomyocytes, or alveolar cells (HPA tissue IHC). | HPA lists these specific cell populations as not detected, so check cell identification and controls; cross-reactivity or endogenous detection activity are possible IHC explanations (HPA tissue IHC; general IHC practice). A tissue-level negative call would overstate these cell-specific observations (HPA tissue IHC). |
| Color spread across cells and surrounding section without clear nuclear boundaries (general IHC practice). | Diffuse background makes CDCA5 localisation difficult to score (general IHC practice). Review the primary-antibody control, blocking, washing, and chromogen development before calling weak staining positive (general IHC practice). |
| No staining in lymph-node germinal-center cells or bone-marrow hematopoietic cells (HPA tissue IHC). | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
💡Expected CDCA5 appearanceCall a positive result when staining is clearly nuclear in an HPA high-staining cell population, such as lymph-node germinal-center cells; isolated membrane signal or diffuse color across HPA not-detected reference cells is suspect (HPA tissue IHC; UniProt Q96FF9; general IHC practice).
| Cell-cycle localisation (UniProt Q96FF9) | CDCA5 associates with nuclear chromatin from S phase through metaphase and is released into cytoplasm after nuclear-envelope breakdown; interpret a cytoplasmic cell in its morphological context (UniProt Q96FF9). |
| Tissue and cell selection (HPA tissue IHC) | HPA reports high staining in named populations, including endometrial glandular cells and cerebellar Purkinje cells, and not-detected staining in named populations such as adipocytes; do not assign one intensity to every cell in a tissue (HPA tissue IHC). |
| IHC antibody evidence (HPA antibodies) | HPA rates HPA076007 Enhanced for IHC and HPA023691 Supported for IHC. These are antibody-specific validation summaries; confirm the pattern with the antibody actually used (HPA antibodies; general IHC practice). |
| What should IF/ICC show? (HPA subcellular) | HPA reports enhanced nucleoplasmic localisation in ICC-IF images from A-431, U2OS, and HAP1. Use that observation as a localisation cross-check, while interpreting paraffin IHC against the tissue IHC record (HPA subcellular; HPA tissue IHC). |
| Topology and processing (UniProt Q96FF9) | The 252-aa sororin chain has no annotated signal peptide, propeptide, or transmembrane segment; a membrane-dominant or extracellular pattern has no support from these annotations (UniProt Q96FF9). These annotations do not predict antigen-retrieval needs (UniProt Q96FF9). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining cells are blank (HPA tissue IHC). | A failed detection step or unsuitable antibody conditions are possible general IHC causes; the supplied sources give no CDCA5-specific fixation effect (general IHC practice; HPA tissue IHC). | Verify the correct cell population, positive-control section, catalog antibody instructions, retrieval run, and detection reagents before revising conditions (HPA tissue IHC; general IHC practice). |
| Nuclei and surrounding tissue look uniformly brown (general IHC practice). | Excess background from antibody or detection conditions can obscure a compartment-specific result (general IHC practice). | Inspect a no-primary control; review blocking, washes, antibody concentration, and chromogen development using the antibody's instructions (general IHC practice). |
| A no-primary control develops color (general IHC practice). | Endogenous enzyme activity or nonspecific detection-reagent binding may contribute, depending on the chromogenic system (general IHC practice). | Check the detection-system controls and its recommended endogenous-activity block; repeat before attributing color to CDCA5 (general IHC practice). |
| A membrane rim dominates while nuclei remain pale (UniProt Q96FF9; HPA tissue IHC). | The pattern conflicts with the reported nuclear IHC profile and no-transmembrane topology; nonspecific staining is possible (HPA tissue IHC; UniProt Q96FF9; general IHC practice). | Compare with a known high-staining cell population and no-primary control, then reassess the antibody and detection conditions (HPA tissue IHC; general IHC practice). |
| Cytoplasmic staining appears in a few cells or in placenta (HPA tissue IHC). | Cytoplasmic IHC is reported in a few tissues; UniProt also describes release after nuclear-envelope breakdown (HPA tissue IHC; UniProt Q96FF9). | Check tissue identity and cell morphology, and compare adjacent nuclear staining and controls before classifying the cytoplasmic signal (HPA tissue IHC; UniProt Q96FF9; general IHC practice). |
| Strong signal appears in HPA not-detected reference cells (HPA tissue IHC). | Misidentified cells, cross-reactivity, or detection background are possible; HPA's entries describe particular cell populations rather than whole-tissue absence (HPA tissue IHC; general IHC practice). | Confirm cell identity on the counterstain, inspect the no-primary control, and compare staining with a reported high-staining population (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CDCA5 staining
M05043 · IHCImmunohistochemical analysis of paraffin-embedded human colon, using CDCA5 Antibody.
M05043 · IFImmunofluorescent analysis of Hela cells, using CDCA5 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000146670-CDCA5/tissue/cerebral+cortex#imid_20881079
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000146670-CDCA5/tissue/colon#imid_20880997
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000146670-CDCA5/tissue/liver#imid_20881006
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000146670-CDCA5/tissue/kidney#imid_20880999
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA023691; green = CDCA5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA023691; green = CDCA5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CDCA5 IHC & IF
🧪Run colon first: endocrine cells should show high CDCA5 staining (HPA: High in colon endocrine cells; selected-SKU caption: paraffin-embedded human colon). Use adipose tissue as the negative tissue because CDCA5 is not detected in adipocytes (HPA: Not detected in adipocytes); on the colon slide, use morphologically identified cells lacking specific nuclear staining as internal negative comparators without assuming every non-endocrine cell is negative (UniProt Q96FF9: nuclear localization; standard IHC practice).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CDCA5 in A-431, U2OS, HAP1, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and clonality-matched isotype controls, plus CDCA5-knockout material or a validated peptide-block control if available (standard IHC practice). For chromogenic colon IHC, quench endogenous peroxidase and assess endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected-SKU paraffin-section caption leaves the fixative unreported (selected-SKU caption). Retrieval dependency is unreported, so optimize antigen retrieval empirically for the catalog antibody (standard IHC practice). Whether frozen sections or IF are easier is unreported; ICC-IF images show enhanced nucleoplasmic localization in A-431, U2OS and HAP1, while endogenous peroxidase can complicate chromogenic interpretation in colon sections (HPA: subcellular localization and ICC-IF image cell lines; standard IHC practice).
HPA tissue IHC evidence for CDCA5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CDCA5 IHC Tips
Troubleshoot CDCA5 chromogenic IHC in paraffin sections by checking retrieval, nuclear localisation, cell identity and staining controls.
How should I retrieve CDCA5 in paraffin sections when nuclear staining is weak?
Use Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Run a known positive tissue section alongside the test section; colon endocrine cells or lymph-node germinal-center cells provide reference populations (HPA: High in those cells). If nuclear signal remains weak, compare a shorter and longer heating interval while keeping buffer and detection conditions fixed (standard IHC practice). Check whether longer heating damages morphology or raises background before adopting it (standard IHC practice). Score chromogen within intact nuclei, since nucleoplasm is the reported principal location (HPA subcellular: Nucleoplasm).
Can fixation explain a negative CDCA5 stain in paraffin sections?
CDCA5-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (M05043 caption: human colon; fixative unstated). Record the fixative, fixation duration and section age for each case, then compare sections processed together using identical retrieval and detection (standard IHC practice). A positive control that stains on the same run helps distinguish a case-specific negative result from a failed staining run (standard IHC practice). If archival and newly prepared sections differ, assess morphology and control staining before attributing the difference to CDCA5 biology (standard IHC practice). Avoid assigning an optimal fixation time without a controlled comparison (supplied evidence: no target-specific fixation data).
Which compartment should contain convincing CDCA5 staining?
Prioritise nuclear staining in intact cells, particularly a nucleoplasmic pattern, when reading chromogenic sections (HPA subcellular: enhanced nucleoplasm; HPA tissue IHC: predominantly nuclear). CDCA5 associates with chromatin from S phase until metaphase and can enter the cytoplasm after nuclear-envelope breakdown (UniProt Q96FF9: subcellular location). Thus, assess mitotic figures separately from interphase nuclei instead of treating every cytoplasmic signal as equivalent (UniProt Q96FF9: cell-cycle-dependent location). Widespread diffuse cytoplasmic colour in quiescent-looking cells warrants a check of background and detection controls (HPA tissue IHC: cytoplasmic expression limited to a few tissues). Compare nuclear boundaries with the counterstain at 40× objective magnification when the pattern is ambiguous (standard IHC practice).
Could an isoform or modified epitope explain uneven CDCA5 staining?
The supplied record annotates 0 isoforms and one 1–252 Sororin chain, so it provides no isoform-specific staining explanation (UniProt Q96FF9: isoforms and processing). It does annotate multiple modified residues, including phosphoserines at positions 21, 33, 35, 75 and 79 (UniProt Q96FF9: modified residues). An epitope’s sensitivity to these modifications cannot be inferred without its mapped sequence and antibody validation (supplied evidence: no epitope mapping). If staining varies across a section, compare retrieval conditions and tissue preservation before proposing modification-dependent recognition (standard IHC practice). Use a separately validated reagent or an orthogonal assay to investigate that hypothesis (standard IHC practice).
How can IF help assess an ambiguous CDCA5 IHC pattern?
Use IF/ICC as a complementary localisation check, while evaluating its antibody validation separately from the catalog antibody’s paraffin-section IHC evidence (M05043 caption: paraffin human colon; HPA subcellular: ICC/IF images). Multiplex CDCA5 with a validated marker for the cell population under study; colon endocrine cells are one supported IHC reference population (HPA tissue IHC: High in colon endocrine cells). Choose fluorophores after inspecting unstained tissue in each channel, because tissue autofluorescence can obscure weak signal (standard IF practice). Permeabilise to expose the nuclear epitope and compare matched permeabilised and minimally permeabilised preparations (UniProt Q96FF9: no transmembrane segment; standard IF practice). Assess at least 2 channels independently before interpreting overlap (standard IF practice).
How do I distinguish CDCA5 signal from chromogenic background?
Examine a no-primary control and the positive control under the same retrieval and detection conditions (standard IHC practice). Include a peroxidase-block step before DAB development, then check whether colour persists without primary antibody (standard chromogenic IHC practice). Diffuse deposits, tissue-edge staining and pigment that appears in the no-primary control should not be scored as CDCA5 (standard IHC practice). Compare nuclear signal in reference populations with a lower-signal comparator: lymph-node germinal-center cells are High, whereas adipocytes are Not detected (HPA tissue IHC). If background dominates, titrate the primary antibody and shorten chromogen development in separate comparisons, preserving the same counterstain and imaging settings (standard IHC practice).
What is a defensible way to quantify CDCA5 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and population before scoring: use nuclear chromogen in morphologically intact cells as the primary readout (HPA subcellular: nucleoplasm; standard IHC practice). Report the percentage of positive nuclei and intensity, or calculate an H-score on a 0–300 scale using prespecified intensity bins (standard IHC practice). For spatial comparisons, report positive nuclei per mm² of viable tissue and document the sampled area (standard IHC practice). Normalise counts to the number of eligible cells or viable tissue area, rather than the entire section area (standard IHC practice). Keep retrieval, staining batch, thresholds and imaging settings consistent across samples (standard IHC practice).
When should a CDCA5-positive IHC result be questioned?
Give greatest weight to staining in intact nuclei within an identified cell population, because nucleoplasm is the enhanced location and most tissue expression is nuclear (HPA subcellular; HPA tissue IHC). Question apparent positives confined to tissue edges, necrotic regions or no-primary controls (standard IHC practice). Check cell identity: colon endocrine cells and lymph-node germinal-center cells are reported High, while adipocytes and cardiomyocytes are Not detected (HPA tissue IHC). Cytoplasmic staining needs context because CDCA5 can redistribute after nuclear-envelope breakdown, and HPA reports cytoplasmic expression in only a few tissues (UniProt Q96FF9: subcellular location; HPA tissue IHC). Resolve disagreement with a matched control section and morphology before calling an isolated signal positive (standard IHC practice).
Boster reagents
Best CDCA5 / Sororin IHC Antibodies
M05043 has human-reactive IHC and IF applications, with images of paraffin-embedded human colon and HeLa cells (catalog: reactivity, applications, image captions).
Real IHC data
Anti-CDCA5/Sororin Rabbit Monoclonal Antibody
Cat # M05043
M05043 will render with an IHC image of paraffin-embedded human colon (M05043 IHC image caption). Its catalog also shows IF staining in HeLa cells (M05043 IF image caption).
Which to pick: For tissue IHC, choose M05043, a rabbit monoclonal shown on paraffin-embedded human colon; the fixative is unreported (catalog: host and clone; M05043 IHC image caption). For IF/ICC, M05043 lists both applications and has a HeLa-cell IF image (catalog: applications; M05043 IF image caption). For other species, the catalog provides no reactivity claim beyond Human (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96FF9 (CDCA5_HUMAN, Sororin).
- Human Protein Atlas. CDCA5 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CDCA5 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. CDCA5 antibody validation summary (2 antibodies).
- CDCA5 overexpression is an Indicator of poor prognosis in patients with hepatocellular carcinoma (HCC). BMC cancer 2018 — PMC6267780.
- Mechanistic and Clinical Evidence Supports a Key Role for Cell Division Cycle Associated 5 (CDCA5) as an Independent Predictor of Outcome in Invasive Breast Cancer. Cancers 2022 — PMC9688237.
- Therapeutic potential of targeting cell division cycle associated 5 for oral squamous cell carcinoma. Oncotarget 2016 — PMC4823039.
- The impact of CDCA5 expression on the immune microenvironment and its potential utility as a biomarker for PD-L1/PD-1 inhibitors in lung adenocarcinoma. Translational oncology 2024 — PMC11214526.
- PubMed PMID:12188893 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CDCA5 (Sororin). Expected localisation Mostly nuclear; cytoplasmic in some tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: M05043 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet M05043); 2. Fixation: Image fixative and duration unreported (datasheet M05043); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit monoclonal (clone IAH-3) anti-CDCA5, 1:50 (datasheet M05043); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Nuclear envelope breakdown can shift CDCA5 into cytoplasm (UniProt); Isoform / epitope: No annotated isoforms; one 1–252 chain (UniProt); Regulation: S phase–metaphase chromatin binding (UniProt). Evidence: UniProt Q96FF9, HPA tissue IHC · Enhanced, HPA ICC-IF · nucleoplasm., Datasheet M05043, PMC6267780, PMC9688237. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CDCA5/immunohistochemistry-ihc-guide.