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NCAPG / Condensin complex subunit 3 · IHC design guide
Design Immunohistochemistry for NCAPG
Plan chromogenic NCAPG IHC on paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A05501-1). Compare staining with the reported cytoplasmic tissue pattern, while accounting for its uncertain reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NCAPG 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 | Cytoplasmic staining in tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05501-1) | |
| Positive control | Cerebellum+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 (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 A05501-1) | |
| Caveat | Presumed off-target staining; verification pending (HPA tissue IHC) | |
| Regulation | Mitotic chromatin association (UniProt) | |
| Isoform / epitope | No annotated isoforms; one 1–1015 chain (UniProt) |
Section 1
Recommended NCAPG IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A05501-1). Published LUAD and glioma IHC methods provide two additional protocols (PMC8380402; PMC8906777).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A05501-1) |
| Fixation | Image fixative and duration unreported (datasheet A05501-1); 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 A05501-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05501-1) |
| Primary antibody | Rabbit anti-NCAPG, 2-5 μg/ml (datasheet A05501-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05501-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05501-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NCAPG-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several different tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A05501-1), then follow the published dilution and detection details for the relevant tissue (PMC8380402; PMC8906777).
Section 2
What Is the Expected NCAPG Staining Pattern?
NCAPG is found in the cytoplasm, nucleus and chromosomes, with chromosome association increasing during mitosis (UniProt Q9BPX3). In tissue IHC, HPA reports cytoplasmic staining across several tissues, including high staining in esophageal squamous epithelial cells and lymph node germinal center cells (HPA tissue IHC). NCAPG has no transmembrane segment (UniProt Q9BPX3 topology). Treat the tissue pattern cautiously: HPA rates its IHC evidence Uncertain because presumed off-target binding was observed (HPA tissue IHC).
| Cell-associated cytoplasmic stain in esophageal squamous epithelial cells or lymph node germinal center cells (HPA tissue IHC). | These are reported high-staining populations, suitable for assessing whether the slide shows the reported tissue pattern (HPA tissue IHC). Cytoplasmic signal is also consistent with the predominantly cytoplasmic condensin complex in interphase cells (UniProt Q9BPX3). Neither match establishes antibody specificity, given HPA’s Uncertain IHC rating (HPA tissue IHC). |
| Stain concentrates on condensed chromosomes in cells with recognizable mitotic morphology (UniProt Q9BPX3). | Chromosome association is plausible during mitosis, when most condensin complex associates with chromatin; association falls in late telophase (UniProt Q9BPX3). Compare stained structures with the nuclear counterstain and cell morphology (general IHC practice). A predominantly cytoplasmic pattern in adjacent interphase cells can still fit the same protein’s cell-cycle behavior (UniProt Q9BPX3). |
| Sharp membrane-only staining or extracellular deposits dominate the section (UniProt Q9BPX3 topology). | This distribution does not fit the reported cytoplasmic, nuclear and chromosome locations or the absence of a transmembrane segment (UniProt Q9BPX3). Consider nonspecific staining or detection artifact before calling it NCAPG; HPA has already flagged presumed off-target binding (HPA tissue IHC). Check whether the signal follows cell boundaries, tissue edges or deposits (general IHC practice). |
| Strong staining appears in adipocytes or skeletal muscle myocytes (HPA tissue IHC). | HPA reports NCAPG as not detected in these cell populations, so strong signal conflicts with its reference IHC pattern (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). Because HPA’s own NCAPG IHC evidence is Uncertain, use this discrepancy as a prompt to check controls, not as proof of the cause (HPA tissue IHC). |
| Diffuse color covers tissue, empty spaces or the slide outside identifiable cells (general IHC practice). | This differs from the cell-associated cytoplasmic expression described by HPA (HPA tissue IHC). Review the no-primary and detection controls for background from the staining system, and assess blocking, washing and reagent concentration (general IHC practice). Diffuse cytoplasm within identifiable cells alone is not a failure criterion: cytoplasmic localization is reported for NCAPG (HPA tissue IHC; UniProt Q9BPX3). |
💡Expected NCAPG appearanceCall a result provisionally positive when identifiable esophageal squamous epithelial or lymph node germinal center cells show clear cell-associated, potentially high cytoplasmic staining, with chromosome-associated signal plausible in mitotic cells (HPA tissue IHC; UniProt Q9BPX3). Membrane-only or extracellular color argues against the expected location (UniProt Q9BPX3 topology); HPA’s Uncertain rating limits specificity claims (HPA tissue IHC).
| Cell-cycle state (UniProt Q9BPX3). | Interphase condensin is mostly cytoplasmic, with a minority associated with chromatin or chromosome foci; during mitosis, most associates with chromatin (UniProt Q9BPX3). Interpret compartment alongside cell morphology rather than requiring every cell to show the same distribution (general IHC practice). |
| Tissue reference pattern (HPA tissue IHC). | HPA reports high staining in several cell populations, including esophageal squamous epithelial cells and lymph node germinal center cells, but not detected in adipocytes and skeletal muscle myocytes (HPA tissue IHC). These are reference observations, not universal guarantees for a new assay (HPA tissue IHC: Uncertain). |
| IHC evidence and antibody validation (HPA tissue IHC; HPA antibodies). | HPA assigns the tissue IHC profile an Uncertain reliability rating, notes presumed off-target binding, and lists HPA039613 as IHC Uncertain (HPA tissue IHC; HPA antibodies). Give concordant morphology and controls more weight than a single stained field (general IHC practice). |
| Topology and processing (UniProt Q9BPX3). | NCAPG has no transmembrane segment, signal peptide or annotated propeptide; its listed chain spans residues 1–1015 (UniProt Q9BPX3 topology and processing). These annotations support a non-surface interpretation. They do not identify an antibody epitope or establish how antigen retrieval affects staining (UniProt Q9BPX3). |
| ICC/IF evidence (HPA subcellular). | HPA provides no main ICC/IF location or cell-line images for NCAPG; it notes that transcript variation correlates with the cell cycle (HPA subcellular). The tissue IHC profile and UniProt localization can guide interpretation, but they do not establish an IF/ICC staining pattern or protocol (HPA tissue IHC; UniProt Q9BPX3). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in esophageal squamous epithelial or lymph node germinal center cells (HPA tissue IHC). | The run may have failed, or the tested preparation may differ from HPA’s reported high IHC pattern (general IHC practice; HPA tissue IHC). | Confirm the cell population is present, then check the staining run with its positive control and review the antibody, detection and retrieval conditions used (general IHC practice). HPA’s Uncertain rating means a negative result alone cannot settle NCAPG abundance (HPA tissue IHC). |
| Staining is confined to cell borders or extracellular material (UniProt Q9BPX3 topology). | That location conflicts with NCAPG’s reported intracellular compartments and lack of a transmembrane segment (UniProt Q9BPX3). Nonspecific signal is possible (general IHC practice). | Compare with a no-primary control, inspect tissue edges and deposits, and reassess specificity before scoring the field (general IHC practice). |
| Adipocytes or skeletal muscle myocytes stain strongly (HPA tissue IHC). | HPA reports these populations as not detected; cross-reactivity or endogenous detection activity may contribute (HPA tissue IHC; general IHC practice). | Check cell identity and a no-primary control, then review the detection-system controls and compare with an HPA high-staining population on the same run where available (general IHC practice; HPA tissue IHC). |
| Color is widespread, including spaces without cells (general IHC practice). | Non-cell-associated background can arise from the staining system or insufficient washing (general IHC practice). | Review no-primary and detection controls, washing and blocking; score only signal tied to identifiable cells (general IHC practice). Keep true cytoplasmic staining in consideration because HPA reports it across tissues (HPA tissue IHC). |
| Some cells show cytoplasm while recognizable mitotic cells show chromosome-associated signal (UniProt Q9BPX3). | Condensin changes distribution through the cell cycle (UniProt Q9BPX3). | Assess each cell against morphology and counterstain before treating the compartment difference as assay inconsistency (general IHC practice). Do not infer antibody specificity from this match alone because HPA rates NCAPG IHC Uncertain (HPA tissue IHC). |
| Can an IF/ICC image be used to confirm the IHC pattern? | HPA lists no NCAPG ICC/IF images or main ICC/IF location (HPA subcellular). | Use the reported tissue IHC observations and UniProt cell-cycle localization as provisional context (HPA tissue IHC; UniProt Q9BPX3). Evaluate IF/ICC evidence in its separate guide; HPA supplies no image-based IF/ICC confirmation here (HPA subcellular). |
Experiment evidence
IHC & IF images showing NCAPG staining
A05501-1 · IHCIHC analysis of NCAPG using anti-NCAPG antibody (A05501-1). NCAPG was detected in a paraffin-embedded section of human tonsil tissue. Heat m
A05501-1 · IFIF analysis of NCAPG using anti-NCAPG antibody (A05501-1) and anti-Beta Tubulin antibody (M01857-3). NCAPG was detected in an immunocytochem
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109805-NCAPG/tissue/cerebral+cortex#imid_11488147
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109805-NCAPG/tissue/colon#imid_11488089
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109805-NCAPG/tissue/liver#imid_11488090
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109805-NCAPG/tissue/kidney#imid_11488075
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NCAPG IHC & IF
🧪Run lymph node first and confirm staining in germinal center cells (HPA: High in germinal center cells); use adipose tissue as the negative tissue and assess adipocytes (HPA: Not detected in adipocytes). On the positive slide, cells without DAB deposit above the counterstain provide an internal background comparison, but the supplied HPA row does not establish which other lymph-node cells are NCAPG-negative (HPA: lymph-node row identifies germinal center cells).
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for NCAPG; derive a cell-line control from the positive tissue's cell type (Purkinje cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and primary-host- and clonality-matched isotype controls, plus an NCAPG knockout specimen if available (standard IHC practice). Quench endogenous peroxidase and inspect the no-primary lymph-node section for DAB background from peroxidase-positive leukocytes (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A05501-1 paraffin-section caption does not state the fixative (selected SKU tissue-IHC caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected SKU tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; HPA lists no cell lines with ICC-IF images, and peroxidase-positive leukocytes in lymph node can complicate DAB interpretation (HPA: subcellular record; standard IHC practice).
HPA tissue IHC evidence for NCAPG
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Presumed off target binding observed and disregarded. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NCAPG IHC Tips
Troubleshoot NCAPG chromogenic IHC by checking retrieval, staining compartment, and tissue context; interpret tissue patterns cautiously because external verification is pending (HPA: reliability Uncertain).
Which retrieval condition should I try first for NCAPG in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A05501-1: tissue IHC caption). The catalog antibody detected NCAPG in a human tonsil paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A05501-1). If staining is weak, compare retrieval heating and cooling conditions across matched sections while holding antibody concentration and detection constant (standard IHC practice). Include a section processed without primary antibody to identify detection background, and record whether changes improve cellular staining without increasing tissue-edge staining (standard IHC practice).
How should I assess whether fixation is affecting NCAPG staining?
Target-specific fixation sensitivity is unknown: the catalog tissue-IHC caption identifies a paraffin section but does not state its fixative (datasheet A05501-1). Record fixative, fixation duration, tissue thickness, and processing history before comparing staining across specimens (standard IHC practice). If archived sections differ in preparation, test matched material processed under documented conditions and keep EDTA pH 8.0 retrieval and detection settings consistent (datasheet A05501-1; standard IHC practice). Interpret a loss of signal as a preparation-associated observation until matched controls separate fixation effects from retrieval, tissue condition, and detection effects (standard IHC practice).
Should NCAPG staining appear in nuclei, cytoplasm, or mitotic chromosomes?
Assess compartment together with cell-cycle state: most condensin complex is cytoplasmic in interphase, while a minority associates with chromatin and chromosome foci (UniProt Q9BPX3: subcellular location). During mitosis, most condensin associates with chromatin, including chromosome arms after prophase onset, and dissociates in late telophase (UniProt Q9BPX3: subcellular location). HPA reports cytoplasmic staining across several tissues, but its tissue-IHC reliability is Uncertain because presumed off-target binding was observed (HPA: tissue IHC). Review chromogenic signal against a nuclear counterstain and cell morphology, and avoid treating diffuse staining in every cell as proof of NCAPG localisation (standard IHC practice; HPA: reliability Uncertain).
How can I investigate discordant NCAPG staining when the antibody epitope is unclear?
Check the catalog antibody’s documented immunogen or epitope before assigning a staining difference to epitope masking; the supplied tissue-IHC caption does not identify that region (datasheet A05501-1). UniProt lists 0 isoforms, no transmembrane segment, and a single 1–1015 chain, so the supplied record does not support an isoform-specific staining explanation (UniProt Q9BPX3). Reported phosphosites include Ser390, Ser674, Thr931, Ser973, and Ser975, but their effect on this antibody’s binding is unreported (UniProt Q9BPX3: modified residues; datasheet A05501-1). Compare matched retrieval conditions and, where available, an independently validated epitope before attributing discordance to modification or processing (standard IHC practice).
How should I adapt NCAPG localisation checks to multiplex immunofluorescence?
For a separate IF experiment, pair NCAPG with a validated marker for the cell population being examined, such as germinal-center cells in lymphoid tissue (HPA: high staining in lymph-node germinal-center cells; standard IF practice). Choose fluorophores and filter sets after measuring tissue autofluorescence, and favor a spectrally separated channel where background is lower (standard IF practice). Because NCAPG is intracellular and has no transmembrane segment, assess permeabilisation for access to cytoplasmic, nuclear, and chromosome-associated antigen (UniProt Q9BPX3: location and topology; standard IF practice). Include single-stain and no-primary controls to evaluate bleed-through and nonspecific fluorescence; the supplied IHC caption does not establish IF performance (standard IF practice; datasheet A05501-1).
What controls help distinguish NCAPG signal from chromogenic background?
Run a no-primary control through the same secondary antibody and DAB workflow, and compare it with stained sections for diffuse deposit or tissue-edge signal (standard IHC practice). The catalog tonsil example used 10% goat serum blocking, a peroxidase-conjugated goat anti-rabbit secondary for 30 minutes at 37°C, and DAB development (datasheet A05501-1). Add an endogenous peroxidase block as a general chromogenic IHC step, particularly when the no-primary control shows enzyme-associated color (standard IHC practice). Titrate detection and primary antibody on matched sections if background persists; HPA’s presumed off-target binding means cytoplasmic color alone cannot establish specificity (standard IHC practice; HPA: reliability Uncertain).
How should I score NCAPG staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because NCAPG distribution changes between interphase and mitosis (UniProt Q9BPX3: subcellular location; standard IHC practice). For chromogenic sections, report the percentage of positive target cells and intensity by compartment, or calculate an H-score from the percentages at 0–3 intensity, yielding 0–300 (standard IHC scoring practice). Normalise positive-cell counts to the number of evaluable target cells, or mitotic-cell counts to evaluable tissue area in mm², and use the same thresholds across sections (standard IHC practice). Exclude damaged regions and document background subtraction, since the HPA tissue-IHC assessment is Uncertain (standard IHC practice; HPA: tissue IHC).
When is an apparent NCAPG-positive cell likely to be artefactual?
Give greatest weight to cellular staining whose compartment and morphology fit cytoplasmic interphase condensin or chromosome-associated mitotic condensin (UniProt Q9BPX3: subcellular location). Scrutinise staining confined to tissue edges, damaged or necrotic regions, or no-primary controls, and check whether endogenous peroxidase contributes to DAB color (standard IHC practice). Unexpected staining across cells without corresponding cellular structure needs independent validation; HPA labels its tissue-IHC profile Uncertain after presumed off-target binding (HPA: tissue IHC). Use the catalog tonsil image as evidence for staining under its stated EDTA pH 8.0 conditions, while treating the unspecified fixative and unverified specificity as limits on interpretation (datasheet A05501-1; HPA: reliability Uncertain).
Boster reagents
Best NCAPG / Condensin complex subunit 3 IHC Antibodies
A05501-1 has IHC data from a human tonsil paraffin section and IF/ICC data from HeLa cells (catalog image captions); its listed reactivity is human (catalog).
Real IHC data
Anti-NCAPG Antibody ®
Cat # A05501-1
A05501-1 is listed for human IHC and IF/ICC (catalog applications and reactivity). Its images show NCAPG staining in a human tonsil paraffin section by IHC and in HeLa cells by IF/ICC (catalog image captions).
Which to pick: Choose A05501-1 for paraffin-section IHC: its own tonsil image used EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (A05501-1 IHC image caption). Choose A05501-1 for IF/ICC at the listed 5 μg/ml; its own IF image shows staining in HeLa cells (catalog IF dilution and A05501-1 IF image caption). No cross-species option is documented here: A05501-1 lists human reactivity, and its clonality is unreported (catalog reactivity and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BPX3 (CND3_HUMAN, Condensin complex subunit 3).
- Human Protein Atlas. NCAPG tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. NCAPG subcellular location (ICC-IF): Variation in transcript expression is correlated to the cell cycle..
- Human Protein Atlas. NCAPG antibody validation summary (1 antibodies).
- Novel candidate metastasis-associated genes for synovial sarcoma. Journal of cellular and molecular medicine 2024 — PMC11267982.
- NCAPG promotes the progression of lung adenocarcinoma via the TGF-β signaling pathway. Cancer cell international 2021 — PMC8380402.
- Elevated mRNA Expression Levels of NCAPG are Associated with Poor Prognosis in Ovarian Cancer. Cancer management and research 2020 — PMC7369365.
- NCAPG as a Novel Prognostic Biomarker in Glioma. Frontiers in oncology 2022 — PMC8906777.
- PubMed PMID:10910072 — UniProt-cited evidence.
- PubMed PMID:11136719 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NCAPG (Condensin complex subunit 3). Expected localisation Cytoplasmic staining in tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05501-1). Validated IHC antibody: A05501-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A05501-1); 2. Fixation: Image fixative and duration unreported (datasheet A05501-1); 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 A05501-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05501-1); 8. Primary antibody: Rabbit anti-NCAPG, 2-5 μg/ml (datasheet A05501-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05501-1). Watch for: Fixation: Keep paraffin-section 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 A05501-1); Caveat: Presumed off-target staining; verification pending (HPA tissue IHC); Isoform / epitope: No annotated isoforms; one 1–1015 chain (UniProt); Regulation: Mitotic chromatin association (UniProt). Evidence: UniProt Q9BPX3, HPA tissue IHC · Uncertain, HPA cell-line RNA · Variation in transcript expression is correlated to the cell cycle., Datasheet A05501-1, PMC8380402, PMC8906777. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NCAPG/immunohistochemistry-ihc-guide.