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CDCA5 / Sororin · IHC design guide
Design Immunohistochemistry for CDCA5
Plan chromogenic CDCA5 IHC around nuclear staining in most tissues (HPA tissue IHC). Use high-staining lymph-node germinal-center cells (HPA tissue IHC) and start catalog antibody M05043 at 1:50 (datasheet).
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 | Predominantly nuclear; cytoplasmic in a few tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in most tissues; placental cytoplasm (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 | Mitotic nuclear-envelope breakdown can shift signal to cytoplasm (UniProt) | |
| Regulation | Chromatin binding varies with cell-cycle stage (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one 1–252 chain (UniProt) |
Section 1
Recommended CDCA5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by 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 retrieval at 95–98 °C for 20 min (page retrieval rule); compare citrate pH 6.0 if needed (PMC9688237; PMC4823039).
Section 2
What Is the Expected CDCA5 Staining Pattern?
CDCA5 is associated with nuclear chromatin from S phase until metaphase and is released into the cytoplasm upon nuclear envelope breakdown (UniProt Q96FF9). In tissue IHC, expect chiefly nuclear staining in selected cells, including bone marrow hematopoietic cells and lymph node germinal center cells (HPA tissue IHC). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC). CDCA5 has no transmembrane segment (UniProt Q96FF9 topology).
| Distinct nuclear staining in bone marrow hematopoietic cells or lymph node germinal center cells. | This fits HPA's High staining in those cell populations and its predominantly nuclear tissue profile (HPA tissue IHC). Score the specified cells and their nuclear compartment; a positive tissue need not show uniform staining across every cell (general IHC interpretation). |
| Predominantly membranous staining, or widespread cytoplasmic staining without the expected nuclear pattern. | Check for an artefact or unrelated binding: CDCA5 has no transmembrane segment, and HPA describes nuclear expression in most tissues (UniProt Q96FF9 topology; HPA tissue IHC). Do not reject every cytoplasmic signal: HPA reports it in a few tissues, including placenta, and UniProt describes cytoplasmic release after nuclear envelope breakdown (HPA tissue IHC; UniProt Q96FF9). |
| Strong signal in prostate glandular cells or adipocytes while a known-positive cell population also stains. | HPA reports CDCA5 as Not detected in those specified cells (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; inspect the signal's compartment and compare an appropriate detection-only control before calling these cells CDCA5 positive (general IHC practice). |
| Diffuse colour over many compartments, tissue structures, or the section background. | A diffuse deposit does not establish the cell-resolved nuclear pattern described by HPA (HPA tissue IHC). Consider nonspecific binding or detection background; assess a no-primary control and review blocking, washes, and detection conditions (general IHC practice). |
| No staining in bone marrow hematopoietic cells or lymph node germinal center cells. | These are HPA High populations, so their absence makes a negative study difficult to interpret (HPA tissue IHC). Check tissue preservation and morphology, antibody and detection setup, and retrieval conditions using the same positive control section (general IHC practice). |
💡Expected CDCA5 appearanceCall a convincing positive when defined cells show clear nuclear signal, potentially strong in HPA High populations such as bone marrow hematopoietic cells; widespread membranous staining or colour in HPA Not detected cell populations warrants investigation before scoring it as CDCA5 (HPA tissue IHC; UniProt Q96FF9 topology).
| Cell cycle and compartment | CDCA5 associates with nuclear chromatin from S phase until metaphase and is released into cytoplasm after nuclear envelope breakdown (UniProt Q96FF9). Interpret an occasional cytoplasmic cell in its tissue context rather than requiring every positive cell to have identical localisation. |
| Antibody evidence | HPA023691 is IHC Supported; HPA076007 is IHC Enhanced (HPA antibodies). These validation labels differ, while the overall HPA tissue profile is Enhanced with medium staining–RNA consistency (HPA tissue IHC). Record the antibody used when comparing sections. |
| Protein topology and processing | UniProt lists one Sororin chain spanning residues 1–252, no signal peptide or propeptide, and no transmembrane segment (UniProt Q96FF9). These annotations support checking a membrane-dominant pattern; they do not establish how paraffin processing or antigen retrieval affects this antibody. |
| IF/ICC Q: What localisation should I expect? | A: HPA reports enhanced nucleoplasmic localisation in ICC-IF, with images from A-431, U2OS, and HAP1 cells (HPA subcellular). Use that as compartment context; this IHC-P section does not specify an IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive tissue has no visible nuclear signal. | The run may have insufficient usable signal; HPA reports High staining in bone marrow hematopoietic and lymph node germinal center cells (HPA tissue IHC). | Confirm the relevant cells are present, then review the antibody, retrieval, and chromogenic detection steps with a positive control on the same run (general IHC practice). |
| Colour appears across the section and obscures cell boundaries. | Nonspecific binding or detection background can prevent assessment of the expected nuclear distribution (general IHC practice; HPA tissue IHC). | Compare a no-primary control; review blocking, washing, and detection conditions before interpreting weak cellular staining (general IHC practice). |
| Signal is chiefly at cell membranes. | That compartment conflicts with CDCA5's lack of a transmembrane segment and HPA's mainly nuclear tissue profile (UniProt Q96FF9 topology; HPA tissue IHC). | Inspect whether the signal persists in a detection-only control and recheck localisation in an HPA High cell population (general IHC practice; HPA tissue IHC). |
| Cytoplasmic signal appears in an otherwise interpretable section. | HPA describes cytoplasmic expression in a few tissues, including placenta; UniProt also describes release into cytoplasm after nuclear envelope breakdown (HPA tissue IHC; UniProt Q96FF9). | Record the tissue and affected cells, assess morphology and nuclear staining, and avoid treating cytoplasmic localisation alone as proof of either specificity or artefact. |
| Prostate glandular cells or adipocytes stain strongly. | Those cell populations are Not detected in HPA tissue IHC; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare a detection-only control and a known-positive population, then reassess the compartment before assigning a CDCA5-positive score (general IHC practice). |
| Two antibodies give different IHC patterns. | Their HPA IHC validation levels differ: HPA023691 is Supported and HPA076007 is Enhanced (HPA antibodies). The labels alone do not identify the cause of a particular discrepancy. | Document each antibody and compare the same cell populations, compartments, and controls; interpret agreement against the HPA tissue profile (HPA antibodies; 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 stain strongly (HPA: High in colon endocrine cells; M05043 tissue-IHC caption: human colon). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the colon slide, use cells lacking convincing CDCA5 staining as internal background comparators without assuming that every non-endocrine cell is negative.
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 a no-primary, secondary-only control; an isotype control matched to the primary antibody’s host species and clonality; and CDCA5 knockout material or a validated peptide-block control (standard IHC practice). For colon chromogenic IHC, block endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A CDCA5-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the fixative for the selected paraffin-section image is also unreported (M05043 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier, although ICC-IF images show enhanced nucleoplasmic localization in A-431, U2OS and HAP1 (HPA: subcellular). In colon, assess staining against local mucin and luminal background before scoring endocrine cells (standard IHC practice; HPA: High in colon endocrine cells).
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 staining by checking retrieval, compartment, cell type and controls before interpreting chromogenic signal.
How should I retrieve CDCA5 in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). If nuclear staining remains weak, compare a modestly longer retrieval time on matched sections while keeping cooling and detection conditions identical (standard IHC practice). Check whether the change improves staining in colon endocrine cells, a reported high-expression population, without increasing diffuse staining elsewhere (HPA: High in colon endocrine cells; standard IHC practice). Record retrieval time and section quality for each comparison; excessive heating can damage morphology and make nuclear scoring unreliable (standard IHC practice).
Could fixation explain weak or patchy CDCA5 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence, and the catalog antibody’s paraffin-section colon caption does not report a fixative (catalog antibody M05043 caption). Compare sections with documented fixation histories under the same pH 9.0, 20 min retrieval condition before assigning a signal difference to fixation (page retrieval rule; standard IHC practice). Inspect nuclear detail, tissue preservation and section adherence alongside staining, since processing defects can produce patchy chromogenic results (standard IHC practice). Include a consistently processed comparison section in each run, and report the actual fixative when known rather than inferring it from tissue staining patterns (standard IHC practice).
Where should convincing CDCA5 staining appear in IHC sections?
Prioritise interpretable nuclear staining: CDCA5 associates with chromatin from S phase until metaphase, while the tissue atlas reports nuclear expression in most tissues (UniProt Q96FF9 localisation; HPA: tissue profile). Cytoplasmic staining can occur in a few tissues, including placenta, and CDCA5 is released into cytoplasm upon nuclear envelope breakdown (HPA: tissue profile; UniProt Q96FF9 localisation). Score nuclear and cytoplasmic signal separately across at least 2 matched sections, retaining cell morphology and the counterstain for review (standard IHC practice). Diffuse cytoplasmic colour without a plausible cell pattern needs comparison with a no-primary control before it is called CDCA5 (standard IHC practice).
How do epitope uncertainty and CDCA5 modifications affect IHC troubleshooting?
The supplied record lists 0 annotated isoforms and a 1–252 Sororin chain, but gives no epitope location for the catalog antibody (UniProt Q96FF9 record; catalog antibody evidence). CDCA5 has annotated phosphorylation sites, including serines 21, 33 and 35; their effect on this antibody’s staining is unknown (UniProt Q96FF9 modified residues; catalog antibody evidence). When staining differs between samples, compare retrieval and antibody titration on matched sections before attributing the difference to an isoform or modification (standard IHC practice). Keep the same detection settings and score compartment-specific staining so an apparent epitope effect is not a processing difference (standard IHC practice).
How should I assess CDCA5 by IF alongside this IHC guide?
For IF/ICC, compare CDCA5 with a marker for the expected cell population, such as a validated endocrine-cell marker when examining colon, where endocrine cells show high IHC expression (HPA: High in colon endocrine cells; standard IF practice). Choose a fluorophore away from the tissue’s strongest autofluorescence and inspect single-channel controls before interpreting overlap (standard IF practice). Because CDCA5 has no transmembrane segment and is reported in the nucleoplasm, permeabilisation must allow antibody access to intracellular and nuclear epitopes (UniProt Q96FF9 topology; HPA: subcellular location; standard IF practice). The supplied evidence gives no IF/ICC fixation comparison or antibody-specific permeabilisation setting, so validate those conditions with matched controls (supplied evidence; standard IF practice).
How can I distinguish CDCA5 signal from chromogenic background?
Run a no-primary section through the same detection sequence to reveal secondary-reagent, endogenous-enzyme or chromogen background (standard IHC practice). For peroxidase-based detection, use an appropriate peroxidase block before DAB development and compare staining after identical 10 min development intervals where the detection system permits (standard IHC practice). Reassess antibody concentration and washing if colour spreads beyond intact cells or accumulates at section edges; these patterns require control comparison (standard IHC practice). Judge residual signal against nuclear staining in an expected population, such as colon endocrine cells, while retaining the hematoxylin counterstain for cell identification (HPA: High in colon endocrine cells; standard IHC practice).
What is a defensible way to quantify CDCA5 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored compartment before analysis, since CDCA5 is predominantly nuclear in the tissue atlas but can show cytoplasmic expression in some tissues (HPA: tissue profile). Report the percentage of positive nuclei or a nuclear H-score using fixed intensity categories from 0–3, with one threshold applied to every section (standard IHC practice). Normalise counts to the number of evaluable cells in the prespecified cell population, or report positive-cell density per mm² of viable tissue (standard IHC practice). Exclude folds, necrosis and detached edges by a documented rule, and compare matched regions and processing batches before drawing biological conclusions (standard IHC practice).
When is a CDCA5-positive IHC result biologically credible?
A credible result has identifiable cellular staining in a plausible compartment: the tissue atlas reports mainly nuclear expression, and UniProt also describes chromatin association and cell-cycle-dependent cytoplasmic release (HPA: tissue profile; UniProt Q96FF9 localisation). In colon, assess the stained cell type against the reported high expression in endocrine cells rather than assigning every coloured cell the same identity (HPA: High in colon endocrine cells; standard IHC practice). Review 2 controls or matched sections when signal is confined to cut edges, necrosis or the wrong compartment (standard IHC practice). Colour persisting without primary antibody may reflect endogenous enzyme activity or detection background and should not be scored as CDCA5 (standard IHC practice).
Boster reagents
Best CDCA5 / Sororin IHC Antibodies
The human-reactive anti-CDCA5 antibody M05043 has IHC data from paraffin-embedded human colon and IF data from HeLa cells (catalog: Human reactivity; IHC and IF image captions).
Real IHC data
Anti-CDCA5/Sororin Rabbit Monoclonal Antibody
Cat # M05043
M05043 is listed for human IHC, with an image caption showing paraffin-embedded human colon (catalog: IHC application and Human reactivity; IHC image caption). The same SKU is listed for ICC/IF, with an IF image caption showing HeLa cells (catalog: ICC/IF applications; IF image caption).
Which to pick: Choose M05043 for paraffin-section tissue IHC because its own image caption shows paraffin-embedded human colon; the fixative is unreported (IHC image caption). Choose M05043 for IF/ICC because it is a rabbit monoclonal antibody listed for both applications and has an IF image of HeLa cells (catalog: host, clonality and applications; IF image caption). Cross-species use is unverified because the catalog lists Human reactivity only (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 Predominantly nuclear; cytoplasmic in a few 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: Mitotic nuclear-envelope breakdown can shift signal to cytoplasm (UniProt); Isoform / epitope: 0 annotated isoforms; one 1–252 chain (UniProt); Regulation: Chromatin binding varies with cell-cycle stage (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.