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NUSAP1 / Nucleolar and spindle-associated protein 1 · IHC design guide
Design Immunohistochemistry for NUSAP1
This guide uses high staining in bone marrow hematopoietic cells and undetected staining in adipocytes as cell-level references for paraffin-section NUSAP1 IHC (HPA tissue IHC). Interpret nuclear immune-cell staining alongside the reported off-target binding caveat (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUSAP1 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 in immune cells; cytoplasmic/membranous elsewhere (HPA tissue IHC) | |
| Staining pattern | Immune-cell nuclei; cytoplasmic/membranous staining in other tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06066-2) | |
| 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 cases. (selected-SKU IHC image A06066-2) | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | Cell-cycle linked; intensity unreported (UniProt) | |
| Isoform / epitope | 7 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended NUSAP1 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet A06066-2). The published IHC protocols below describe NUSAP1 staining in kidney and breast cancer samples (PMC11480612; PMC8667124; PMC4618922).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A06066-2) |
| Fixation | Image fixative and duration unreported (datasheet A06066-2); 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 A06066-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06066-2) |
| Primary antibody | Rabbit anti-NUSAP1, 2-5 μg/ml recommended; image 1:100 (datasheet A06066-2) |
| Primary incubation | Overnight at 4 °C (datasheet A06066-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06066-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUSAP1-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in several tissues, most abundant in the nuclear subcellular location of immune cells. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A06066-2). Use the stated citrate retrieval when reproducing either published method (PMC11480612; PMC4618922).
Section 2
What Is the Expected NUSAP1 Staining Pattern?
NUSAP1 can occupy the cytoplasm and nucleolus between divisions, then associate with chromosomes and the mitotic spindle during division (UniProt Q9BXS6). In tissue IHC, expect cell-specific staining, including strong staining in bone marrow hematopoietic cells (HPA: High). HPA rates its tissue pattern Supported, with medium agreement with RNA data and a warning about possible off-target staining (HPA: Supported). NUSAP1 has no transmembrane segment (UniProt Q9BXS6 topology).
| Strong staining in bone marrow hematopoietic cells, with staining in nuclei of immune cells. | This fits an HPA high-staining cell population and its description of nuclear staining as most abundant in immune cells (HPA: High in bone marrow hematopoietic cells; HPA: tissue IHC profile). Record the stained cell type and compartment: intensity alone is insufficient because HPA also reports cytoplasmic and membranous staining in several tissues (HPA: tissue IHC profile). |
| Signal follows a membrane rim alone, without convincing nuclear or division-associated staining. | Review the compartment call and controls before assigning this to NUSAP1. UniProt places NUSAP1 in the cytoplasm, nucleolus, spindle and chromosomes and reports no transmembrane segment (UniProt Q9BXS6). HPA nevertheless describes membranous staining in several tissues and warns of presumed off-target binding, so a rim is a concern rather than proof of artefact (HPA: tissue IHC profile). |
| A cell type reported as unstained shows strong chromogenic signal. | For example, HPA reports adipocytes as Not detected (HPA: adipose tissue). Check cell identification, primary-antibody controls and the detection system before calling the signal NUSAP1; cross-reactivity is plausible because HPA warns that antibodies target proteins from more than one gene and presumed off-target binding was observed (HPA: tissue IHC reliability). Endogenous detection activity is another general IHC possibility (standard IHC practice). |
| Brown deposit is widespread across cells or tissue spaces, obscuring cell boundaries. | Treat the pattern as background until it can be separated from cell-specific staining. Compare a no-primary control, washing and detection steps, and assess whether a recognizable nuclear or division-associated distribution remains (standard IHC practice; UniProt Q9BXS6 localization). HPA's Supported rating has only medium consistency with RNA data and includes an off-target warning (HPA: tissue IHC reliability). |
| No signal appears in a section containing bone marrow hematopoietic cells. | First confirm that the expected cells are present and that the detection run worked with appropriate controls (standard IHC practice). HPA rates these cells High, so absence merits review of the IHC-validated antibody, retrieval conditions and detection steps (HPA: bone marrow hematopoietic cells High; standard IHC practice). The supplied sources do not establish NUSAP1-specific fixation sensitivity. |
💡Expected NUSAP1 appearanceCall a result convincing when identifiable HPA-high cells, such as bone marrow hematopoietic cells, stain strongly with a plausible nuclear or division-associated distribution; broad deposit or isolated membrane rims warrant control review, especially given HPA's off-target warning (HPA: High in bone marrow hematopoietic cells; HPA: tissue IHC profile and reliability; UniProt Q9BXS6 localization).
| Cell identity and tissue selection | HPA reports High staining in bone marrow hematopoietic cells, testis preleptotene spermatocytes and skin spinous-layer cells, among others (HPA: tissue IHC). It reports adipocytes and cardiomyocytes as Not detected (HPA: tissue IHC). Score the specified cells, since a tissue name alone does not define the expected result. |
| Cell-cycle stage | Interphase localization includes cytoplasm and nucleolus; during mitosis NUSAP1 redistributes to spindle and chromosome-associated regions (UniProt Q9BXS6, partly By similarity). A section with few dividing cells may offer few interpretable spindle profiles. Do not require every positive cell to display a visible spindle. |
| Antibody evidence | HPA calls the tissue IHC pattern Supported, describes medium agreement with RNA data, and warns that antibodies target proteins from more than one gene (HPA: tissue IHC reliability). HPA042904 is IHC Supported, whereas no IHC status is supplied for HPA074847 (HPA: antibody validation). Interpret unexpected staining with controls. |
| Epitope and molecular forms | UniProt lists seven isoforms and multiple modified residues, but the payload gives no antibody epitope or isoform coverage (UniProt Q9BXS6). Those annotations cannot establish which forms the IHC-validated antibody detects or predict a retrieval condition. Validate any change in staining against controls rather than attributing it to a specific modification. |
| IF/ICC: where should fluorescence appear? | HPA reports approved nucleoplasm, nucleoli and nucleoli fibrillar-center localization in ICC-IF, with a warning that the evidence uses antibodies targeting proteins from multiple genes (HPA: subcellular ICC-IF). This answers the localization question for IF/ICC; interpret paraffin-section chromogenic staining against the tissue IHC evidence (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| The expected positive cells are absent from the field. | The selected field cannot test the stated cell-level expectation; HPA's High call applies to specified cells (HPA: tissue IHC). | Locate and document the relevant cells before judging staining. For example, identify hematopoietic cells when using bone marrow (HPA: High in bone marrow hematopoietic cells). |
| Expected positive cells are present, but staining is absent. | A failed staining or detection step is possible (standard IHC practice); the sources give no NUSAP1-specific fixation-effect evidence. | Check run controls, antibody application, retrieval and chromogenic detection using the established IHC workflow (standard IHC practice). Reassess the known-positive cells (HPA: bone marrow hematopoietic cells High). |
| Signal appears mainly as a crisp membrane outline. | NUSAP1 lacks a transmembrane segment, although HPA describes membranous tissue staining and possible off-target binding (UniProt Q9BXS6 topology; HPA: tissue IHC profile and reliability). | Review the same field for nuclear or division-associated staining and compare controls before assigning the outline to NUSAP1 (UniProt Q9BXS6 localization; standard IHC practice). |
| Reported negative cells stain strongly. | Cross-reactivity is plausible under HPA's off-target warning; endogenous chromogenic activity is a separate general possibility (HPA: tissue IHC reliability; standard IHC practice). | Verify the cell type, compare a no-primary control and review detection blocking. Use HPA's Not detected calls for the specified cells as a reference, not a whole-tissue exclusion (HPA: tissue IHC). |
| Diffuse color obscures the expected compartments. | Nonspecific reagent retention or detection background can impair interpretation (standard IHC practice). | Inspect no-primary and run controls, washing and detection conditions; score NUSAP1 only where cells and compartments remain discernible (standard IHC practice; UniProt Q9BXS6 localization). |
| Staining intensity varies among adjacent positive cells. | NUSAP1 localization changes with cell-cycle stage, and HPA assigns staining levels to particular cell types (UniProt Q9BXS6; HPA: tissue IHC). | Record cell identity and compartment alongside intensity; examine visible dividing cells for spindle or chromosome-associated signal without requiring every cell to show it (UniProt Q9BXS6, partly By similarity). |
Experiment evidence
IHC & IF images showing NUSAP1 staining
A06066-2 · IHCIHC analysis of NUSAP1 using anti-NUSAP1 antibody (A06066-2). NUSAP1 was detected in a paraffin-embedded section of human prostate cancer ti
A06066-2 · IHCIHC analysis of NUSAP1 using anti-NUSAP1 antibody (A06066-2). NUSAP1 was detected in a paraffin-embedded section of human stomach cancer tis
A06066-2 · IFIF analysis of NUSAP1 using anti-NUSAP1 antibody (A06066-2) and anti-Beta Tubulin antibody (M01857-3). NUSAP1 was detected in an immunocytoc
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000137804-NUSAP1/tissue/cerebral+cortex#imid_12147417
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000137804-NUSAP1/tissue/colon#imid_12147327
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000137804-NUSAP1/tissue/liver#imid_12147328
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000137804-NUSAP1/tissue/kidney#imid_12147345
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042904; green = NUSAP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042904; green = NUSAP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUSAP1 IHC & IF
🧪Run colon first: endocrine cells should stain (HPA: High in colon endocrine cells). Use heart muscle cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); on the colon slide, cells used to judge background should show counterstain without specific DAB deposition, but their NUSAP1-negative status is not established by the supplied HPA row.
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 NUSAP1 in A-431, U-251MG, U2OS, SiHa, with annotated localisation: Nucleoplasm (approved), Nucleoli (approved), Nucleoli fibrillar center (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype-matched rabbit IgG control for the rabbit primary used in the selected image (caption: rabbit anti-NUSAP1). Confirm specificity with NUSAP1 knockout material where available; for HRP/DAB detection, block endogenous peroxidase in the positive tissue (caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A06066-2 paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The demonstrated IHC procedure used heat retrieval in EDTA at pH 8.0, so use that as a starting condition without treating retrieval dependence as established (selected-SKU caption: EDTA retrieval). The supplied evidence does not establish whether frozen sections or IF are easier, or identify a tissue-specific artefact (supplied HPA and selected-SKU evidence).
HPA tissue IHC evidence for NUSAP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. Presumed off target binding observed and disregarded.). 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 → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Rectum | 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 → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUSAP1 IHC Tips
Use the catalog antibody’s paraffin section evidence to set up NUSAP1 IHC, then assess compartment, cell type, and staining controls together.
How should I retrieve NUSAP1 in paraffin sections when staining is weak?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06066-2). The product image establishes the buffer and pH, but gives no heating time, so start with 15–20 minutes and cool in buffer for 20 minutes as controlled IHC optimization steps (standard IHC practice). Run a previously staining section alongside each retrieval condition to distinguish retrieval failure from a detection problem (standard IHC practice). If signal remains weak, adjust heating time in small steps while holding section thickness, antibody incubation, and DAB development constant (standard IHC practice). Check nuclear and nucleolar detail for tissue damage before accepting stronger color as improved NUSAP1 detection (UniProt Q9BXS6 localization; standard IHC practice).
Does fixation explain weak or uneven NUSAP1 staining?
The paraffin section caption does not state its fixative, and target-specific fixation sensitivity is unknown (datasheet A06066-2). Record the fixative, fixation duration, processing schedule, and section age for each specimen before attributing weak staining to fixation (standard IHC practice). Compare specimens processed together, using the documented EDTA pH 8.0 retrieval and 1:100 primary antibody incubation overnight at 4°C as reference conditions (datasheet A06066-2). If staining varies within a block, examine morphology and compare deeper sections for processing gradients or lost tissue (standard IHC practice). Do not treat the published tissue staining pattern as evidence that a particular fixative preserves this epitope (HPA tissue IHC; datasheet A06066-2).
Which compartments should show convincing NUSAP1 staining?
Assess nucleoplasm and nucleoli separately: both are approved subcellular locations, although the supporting antibodies carry a multiple-gene targeting caution (HPA subcellular). NUSAP1 is reported in cytoplasm and nucleoli during interphase and at the mitotic spindle during mitosis, with the detailed redistribution inferred by similarity (UniProt Q9BXS6). In chromogenic sections, compare staining with nuclear morphology and a hematoxylin counterstain before assigning a spindle pattern (UniProt Q9BXS6; standard IHC practice). Keep cytoplasmic staining in a separate score because the tissue survey also reports cytoplasmic and membranous signal (HPA tissue IHC). Diffuse membrane-only color warrants antibody specificity and background checks because NUSAP1 has no annotated transmembrane segment (UniProt Q9BXS6).
Could epitope choice explain discordant NUSAP1 IHC results?
NUSAP1 has 7 annotated isoforms, so request the antibody’s immunogen coordinates before deciding which variants its staining can represent (UniProt Q9BXS6; standard IHC practice). Its annotated chain spans residues 1–441, with no signal peptide or propeptide, so cleavage should not be assumed when mapping the epitope (UniProt Q9BXS6). The record lists 16 modified residues, including phosphorylation at residues 124, 135, 182, 240, and 244 (UniProt Q9BXS6). If the immunogen overlaps a variant-specific or modified region, compare staining with an independently mapped antibody and matched tissue controls (standard IHC practice). A shared nuclear pattern alone cannot establish isoform recognition or NUSAP1 specificity, particularly given the tissue survey’s off-target caution (HPA tissue IHC).
How can IF help investigate ambiguous chromogenic NUSAP1 staining?
Use IF as a separate localization check when chromogenic staining obscures nucleolar or mitotic detail (UniProt Q9BXS6; standard IF practice). Multiplex NUSAP1 with a marker identifying the expected cell population, such as CD45 for immune cells, because the tissue survey reports prominent nuclear signal in immune cells (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore when the specimen has strong shorter-wavelength autofluorescence, and collect unstained and secondary-only controls (standard IF practice). Because NUSAP1 is intracellular and has no annotated transmembrane segment, optimize mild permeabilisation to access nuclear and spindle epitopes while preserving structure (UniProt Q9BXS6; standard IF practice). Interpret apparent colocalization cautiously: the available subcellular images carry a multiple-gene antibody caution (HPA subcellular).
How do I distinguish NUSAP1 signal from chromogenic background?
The product image used 10% goat serum block, 1:100 primary antibody overnight at 4°C, and HRP with DAB (datasheet A06066-2). If widespread brown signal appears, repeat with a primary-omission control and inspect whether it tracks tissue edges, folds, pigment, or damaged regions (standard IHC practice). Include an endogenous peroxidase block before HRP detection, then compare a shorter DAB development with the original section (standard IHC practice). Titrate primary antibody around the documented dilution while keeping retrieval at EDTA pH 8.0 and detection conditions matched (datasheet A06066-2; standard IHC practice). Persistent staining in implausible compartments merits an independent antibody because presumed off-target binding was reported in the tissue survey (HPA tissue IHC).
What should I score when comparing NUSAP1 across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record nuclear and cytoplasmic staining separately (UniProt Q9BXS6 localization; standard IHC practice). For nuclear signal, use an H-score of 0–300 from the percentages at intensity grades 0–3, or report the percentage of positive target cells (standard IHC practice). If cell numbers vary greatly, report positive-cell density per mm² of viable tissue and normalize positive counts to all evaluable cells of the same type (standard IHC practice). Exclude folds, necrosis, and section edges using consistent rules across slides (standard IHC practice). Record mitotic cells separately, since reported redistribution to the spindle can change the apparent compartment and intensity (UniProt Q9BXS6).
When is a positive NUSAP1 IHC result credible?
A credible result combines suitable morphology with nuclear or nucleolar staining in interphase cells, while any proposed mitotic spindle pattern needs visible mitotic figures (UniProt Q9BXS6; standard IHC practice). Check the cell assignment against the tissue survey: hematopoietic cells in bone marrow were scored high, whereas adipocytes were not detected (HPA tissue IHC). Treat strong membrane-only staining or color concentrated at cut edges and necrotic areas as a prompt for controls, rather than immediate evidence of NUSAP1 expression (UniProt Q9BXS6 topology; standard IHC practice). Primary-omission and peroxidase-block controls help identify detection background and endogenous enzyme signal (standard IHC practice). Concordance remains provisional because the tissue survey reports medium RNA–staining consistency and presumed off-target binding (HPA tissue IHC).
Boster reagents
Best NUSAP1 / Nucleolar and spindle-associated protein 1 IHC Antibodies
A06066-2 has IHC images of human prostate and stomach cancer paraffin sections, plus IF images of U2OS cells and human stomach cancer paraffin sections (catalog image captions).
Real IHC data
Anti-NUSAP1 Antibody ®
Cat # A06066-2
A06066-2 is listed for human IHC and has prostate and stomach cancer paraffin-section images (catalog applications/reactivity; IHC image captions). The same SKU is listed for IF/ICC and has IF images of U2OS cells and human stomach cancer paraffin sections (catalog applications; IF image captions).
Which to pick: Choose A06066-2 for human tissue IHC: its rabbit antibody is listed at 2–5 μg/ml, and its own IHC captions show paraffin sections; the fixative is unreported (catalog host/reactivity/dilution; IHC image captions). For IF/ICC, A06066-2 is listed at 5 μg/ml and has U2OS-cell and human tissue IF images (catalog applications/dilution; IF image captions). For cross-species work, its declared reactivity is Human, although separate IHC captions show mouse testis and epididymis (catalog reactivity; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BXS6 (NUSAP_HUMAN, Nucleolar and spindle-associated protein 1).
- Human Protein Atlas. NUSAP1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. NUSAP1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm, nucleoli and nucleoli fibrillar center. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. NUSAP1 antibody validation summary (2 antibodies).
- The Overexpression of NUSAP1 and GTSE1 Could Predict An Unfavourable Prognosis and Shorter Disease Free Survival in ccRenal Cell Carcinoma. Asian Pacific journal of cancer prevention : APJCP 2024 — PMC11480612.
- HBV core protein enhances WDR46 stabilization to upregulate NUSAP1 and promote HCC progression. Hepatology communications 2025 — PMC12055171.
- NUSAP1 promotes the metastasis of breast cancer cells via the AMPK/PPARγ signaling pathway. Annals of translational medicine 2021 — PMC8667124.
- High Levels of Nucleolar Spindle-Associated Protein and Reduced Levels of BRCA1 Expression Predict Poor Prognosis in Triple-Negative Breast Cancer. PloS one 2015 — PMC4618922.
- PubMed PMID:11483580 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUSAP1 (Nucleolar and spindle-associated protein 1). Expected localisation Nuclear in immune cells; cytoplasmic/membranous elsewhere (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06066-2). Validated IHC antibody: A06066-2 at 2-5 μg/ml recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A06066-2); 2. Fixation: Image fixative and duration unreported (datasheet A06066-2); 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 A06066-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06066-2); 8. Primary antibody: Rabbit anti-NUSAP1, 2-5 μg/ml recommended; image 1:100 (datasheet A06066-2); 9. Primary incubation: Overnight at 4 °C (datasheet A06066-2). Watch for: Fixation: Keep paraffin-section fixation consistent across cases. (selected-SKU IHC image A06066-2); Caveat: Presumed off-target binding may confound staining (HPA tissue IHC); Isoform / epitope: 7 isoforms; epitope coverage is unspecified (UniProt); Regulation: Cell-cycle linked; intensity unreported (UniProt). Evidence: UniProt Q9BXS6, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm, nucleoli and nucleoli fibrillar…, Datasheet A06066-2, PMC11480612, PMC8667124. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUSAP1/immunohistochemistry-ihc-guide.