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NUMA1 / Nuclear mitotic apparatus protein 1 · IHC design guide
Design Immunohistochemistry for NUMA1
Plan NUMA1 staining in paraffin sections around the general nuclear pattern reported in tissue IHC (HPA tissue IHC). The guide covers fixation, controls, and scoring, including how mitotic relocalization may affect interpretation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUMA1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear staining in tissue sections (HPA tissue IHC) | |
| Staining pattern | General nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02018-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02018-1) | |
| Caveat | Mitotic relocalization can affect nuclear scoring (UniProt) | |
| Regulation | Cell-cycle-dependent redistribution (UniProt) | |
| Isoform / epitope | 5 isoforms; confirm epitope coverage (UniProt) |
Section 1
Recommended NUMA1 IHC & IF Protocols
The catalog antibody uses heat-mediated EDTA retrieval at pH 8.0 (datasheet A02018-1). The published IHC protocols below cover Wilms tumors, mouse tissue, and ovarian cancer specimens (PMC6925259; PMC8686194; PMC12537492).
| Sample | Paraffin-embedded human acinar adenocarcinoma of prostate tissue; fixative not specified (datasheet A02018-1) |
| Fixation | Image fixative and duration unreported (datasheet A02018-1); 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 A02018-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02018-1) |
| Primary antibody | Rabbit anti-NUMA1, 1-2 μg/ml (datasheet A02018-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02018-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02018-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUMA1-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A02018-1); Tris-EDTA at pH 9.0 is a published alternative (PMC12537492: Methods).
Section 2
What Is the Expected NUMA1 Staining Pattern?
NUMA1 is predominantly nuclear in interphase and relocates to spindle poles and the polar cell cortex during mitosis (UniProt Q14980 localization). In paraffin section IHC, expect nuclear staining across many cell types, including adipocytes, glandular cells, hematopoietic cells and respiratory epithelial cells reported as strongly positive (HPA: general nuclear expression; High in these cell types). HPA rates its tissue pattern Enhanced, with high consistency between antibody staining and RNA expression (HPA: reliability Enhanced).
| Clear nuclear staining in adipocytes, glandular cells, hematopoietic cells or respiratory epithelium. | This matches the reported general nuclear pattern and High staining in those cell types (HPA: tissue IHC). Judge the cell nucleus against its counterstain; staining need not be identical in every cell (standard IHC practice). |
| Predominantly cytoplasmic or widespread membrane staining in cells with intact nuclei. | Treat this as a compartment mismatch for interphase cells and investigate artefact or antibody specificity (HPA: general nuclear expression; UniProt Q14980: interphase nuclear matrix). A limited cortical pattern in a mitotic cell has a different interpretation (UniProt Q14980: mitotic relocalization). |
| Strong staining confined to an unexpected cell population, with little staining in an HPA positive population. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice). HPA reports low tissue specificity and supplies no negative tissue in this payload, so an unlisted cell type alone cannot be called NUMA1 negative (HPA: RNA specificity; tissue IHC). |
| Haze across nuclei, cytoplasm and surrounding tissue obscures cell boundaries. | Diffuse background prevents a reliable nuclear call (standard IHC practice). Compare with an appropriate detection control, then reassess blocking, antibody concentration and wash conditions; the supplied sources do not identify a NUMA1 specific cause (standard IHC practice). |
| No nuclear signal in a section containing an HPA positive cell population. | A negative result conflicts with the reported High staining in that population (HPA: tissue IHC). First establish that the run's positive control worked before interpreting the specimen as biologically negative (standard IHC practice). |
💡Expected NUMA1 appearanceCall an IHC result positive when identifiable nuclei show clear staining in an HPA reported positive cell population, potentially High in the listed examples; diffuse extranuclear color without a readable nuclear pattern is suspect (HPA: tissue IHC; standard IHC interpretation).
| Cell cycle stage (UniProt Q14980: localization). | Interphase NUMA1 occupies the nuclear matrix; mitotic NUMA1 can appear at spindle poles and the polar cortex (UniProt Q14980: localization). Interpret an isolated extranuclear mitotic figure in its cell cycle context rather than applying the interphase pattern to it. |
| Antibody validation (HPA: antibody records). | HPA019841, HPA019859 and HPA029912 each have Enhanced IHC status (HPA: antibody records). This supports the reported pattern, but the status does not validate an unrelated catalog antibody or establish its dilution or retrieval conditions. |
| Isoforms and epitope coverage (UniProt Q14980: five isoforms). | UniProt lists 5 NUMA1 isoforms (UniProt Q14980: isoforms). The supplied record gives no antibody epitope, so it cannot predict which isoforms a particular stain detects or whether isoform coverage explains a weak result. |
| Antigen retrieval and detection controls (standard IHC practice). | Retrieval and chromogenic detection settings can affect whether an IHC run is interpretable (standard IHC practice). Neither HPA tissue staining nor UniProt topology establishes NUMA1 specific fixation sensitivity or an optimal retrieval setting (HPA: tissue IHC; UniProt Q14980: topology). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a known positive control tissue. | An unsuccessful IHC run is possible; HPA reports High staining in several listed cell populations (HPA: tissue IHC). The supplied sources do not identify which workflow step failed. | Check section integrity, reagent activity, antibody use conditions, retrieval and chromogen development against the run controls (standard IHC practice). Reassess specimen negativity only after the positive control works. |
| Uniform brown background obscures nuclear detail. | Nonspecific antibody binding or detection background may obscure the expected nuclear pattern (standard IHC practice; HPA: general nuclear expression). | Inspect a control omitting primary antibody, then adjust blocking, antibody concentration or washes as indicated by that control (standard IHC practice). Record whether the background is nuclear, cytoplasmic or extracellular. |
| Brown signal appears in unexpected cells or structures. | Cross-reactivity or endogenous peroxidase activity may contribute to chromogenic signal (standard IHC practice). HPA provides no negative cell list here, so unexpected cell identity alone does not prove artefact (HPA: tissue IHC). | Use a primary omission control to assess detection activity and an independent antibody or other specificity control if available (standard IHC practice; HPA: Enhanced IHC antibody records). |
| Interphase cells show chiefly cytoplasmic staining. | This conflicts with the predominant nuclear pattern (HPA: tissue IHC; UniProt Q14980: interphase nuclear matrix). A mitotic figure can instead show extranuclear NUMA1 (UniProt Q14980: localization). | Check nuclear morphology and mitotic state, then compare the compartment pattern with a working positive control and detection control before assigning the signal to NUMA1 (standard IHC practice). |
| A few dividing cells show spindle pole or cortical signal. | NUMA1 moves from the interphase nucleus to spindle poles and polar cortex during mitosis (UniProt Q14980: localization). | Interpret these cells separately from the surrounding interphase nuclei; use recognizable mitotic morphology to support the assignment (UniProt Q14980: localization; standard IHC interpretation). |
| IF/ICC Q&A: What localization should be expected? | HPA reports NUMA1 mainly in the nucleoplasm, with an enhanced subcellular location assignment (HPA: ICC-IF subcellular). UniProt also describes cell cycle dependent relocalization (UniProt Q14980: localization). | Expect predominantly nucleoplasmic signal in interphase cells; assess apparent spindle pole or cortical signal in the context of mitosis (HPA: ICC-IF subcellular; UniProt Q14980: localization). |
Experiment evidence
IHC & IF images showing NUMA1 staining
A02018-1 · IHCIHC analysis of NUMA/NUMA1 using anti-NUMA/NUMA1 antibody (A02018-1). NUMA/NUMA1 was detected in a paraffin-embedded section of human acinar
A02018-1 · IHCIHC analysis of NUMA/NUMA1 using anti-NUMA/NUMA1 antibody (A02018-1). NUMA/NUMA1 was detected in a paraffin-embedded section of human breast
A02018-1 · IFIF analysis of NUMA/NUMA1 using anti-NUMA/NUMA1 antibody (A02018-1) and anti-Beta Tubulin antibody (M01857-3). NUMA/NUMA1 was detected in im
M02018-1 · IHCIHC analysis of NUMA using anti-NUMA antibody (M02018-1). NUMA was detected in a paraffin-embedded section of human bladder cancer tissue. H
M02018-1 · IHCIHC analysis of NUMA using anti-NUMA antibody (M02018-1). NUMA was detected in a paraffin-embedded section of human breast cancer tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000137497-NUMA1/tissue/cerebral+cortex#imid_5277672
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000137497-NUMA1/tissue/colon#imid_5311238
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000137497-NUMA1/tissue/liver#imid_5311223
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000137497-NUMA1/tissue/pancreas#imid_5311174
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019841; green = NUMA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019841; green = NUMA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUMA1 IHC & IF
🧪Run breast tissue first and look for nuclear staining in glandular cells (HPA: High in breast glandular cells; UniProt Q14980: interphase nuclear matrix). HPA detects NUMA1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls (HPA: no negative rows). Any cells without specific staining on the positive slide should show counterstained nuclei without DAB; do not assign an unstained cell type in advance (HPA: no negative rows).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: NUMA1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUMA1 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, scrambled (10x), U2OS, siRNA 2 (10x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit IgG isotype control, consistent with the rabbit primary in the selected caption; use NUMA1 knockout material or peptide blocking if a suitable antigen peptide is available as a biological specificity control (selected SKU A02018-1 caption: rabbit primary; standard IHC control practice). Quench endogenous peroxidase and check for pigment that could be mistaken for DAB signal (selected SKU A02018-1 caption: peroxidase detection with DAB; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (selected SKU A02018-1 caption: fixative not stated). The documented IHC conditions use heat retrieval in EDTA at pH 8.0, followed by 2 μg/ml primary antibody overnight at 4°C; retrieval under other conditions requires optimization (selected SKU A02018-1 caption). The supplied evidence does not establish that frozen sections or IF are easier; IF can assess nucleoplasmic localisation, while mitotic cells may show spindle-pole or cortical signal (HPA: nucleoplasm enhanced in ICC-IF; UniProt Q14980: cell-cycle-dependent localisation).
HPA tissue IHC evidence for NUMA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: NUMA1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced NUMA1 IHC Tips
Use the catalog antibody’s paraffin-section workflow as the starting point, then judge NUMA1 staining against its cell-cycle-dependent localisation (datasheet A02018-1; UniProt Q14980).
Which retrieval conditions should I use when NUMA1 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A02018-1). That condition was used for the catalog antibody on a paraffin-embedded human acinar adenocarcinoma of prostate section, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A02018-1). If nuclear staining remains weak, check heating consistency and tissue retention before changing conditions (standard IHC practice). Compare any fallback retrieval condition on adjacent sections with the same antibody concentration and detection steps, using nuclear signal and background together to judge improvement (standard IHC practice).
Could fixation explain weak or uneven NUMA1 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet A02018-1). Record each specimen’s fixative and fixation duration, then compare sections processed with the same EDTA pH 8.0 retrieval and detection workflow (datasheet A02018-1; standard IHC practice). Examine poorly stained areas for uneven preservation or tissue damage before attributing their appearance to NUMA1 biology (standard IHC practice). Keep antibody concentration, retrieval heating and development consistent across the comparison; the documented primary concentration is 2 μg/ml overnight at 4°C (datasheet A02018-1).
Where should NUMA1 staining appear across interphase and mitosis?
Expect predominantly nuclear staining in interphase cells: NUMA1 resides in the nuclear matrix, and tissue IHC shows general nuclear expression (UniProt Q14980 localisation; HPA: general nuclear expression). In dividing cells, examine spindle poles and polar cortex as well as nuclei, because NUMA1 relocalises during mitosis (UniProt Q14980 localisation). Chromogenic paraffin sections may not resolve every spindle structure, so assess mitotic morphology alongside the staining pattern (standard IHC interpretation). If a field shows diffuse cytoplasmic colour without convincing nuclei or identifiable mitotic cells, review retrieval, background and section quality before calling it NUMA1-positive (UniProt Q14980 localisation; standard IHC practice).
Can this antibody distinguish NUMA1 isoforms or modified epitopes?
NUMA1 has 5 listed isoforms and numerous annotated modified residues, including phosphorylation sites (UniProt Q14980 isoforms and modified residues). The supplied caption identifies an IHC staining workflow but does not map the catalog antibody’s epitope or establish isoform selectivity (datasheet A02018-1). Treat a positive nucleus as evidence of staining with this antibody, without assigning a particular isoform or phosphorylation state (standard IHC interpretation). If an isoform-specific conclusion matters, obtain epitope information and compare appropriately characterised specimens or an independently validated reagent; keep the paraffin-section retrieval at EDTA pH 8.0 when comparing this antibody’s staining (datasheet A02018-1; standard IHC practice).
How should I investigate the NUMA1 pattern with IF?
Use IF as a complementary localisation check when chromogenic IHC cannot clearly separate nuclei from nearby mitotic structures (standard IHC/IF practice). Pair NUMA1 with a marker for the expected cell type, such as a glandular epithelial marker in glandular regions, and inspect nuclear counterstaining (HPA: high staining in several glandular cell populations; standard IF practice). Choose fluorophores after checking tissue autofluorescence, favouring a spectrally separated far-red channel when background is strong (standard IF practice). Permeabilise sufficiently to access nuclear NUMA1, then compare nuclear and mitotic patterns; NUMA1 has no annotated transmembrane segment and relocalises to spindle poles and cortex during mitosis (UniProt Q14980 topology and localisation).
What should I check when DAB obscures NUMA1 nuclei?
Check a section without primary antibody to assess secondary-reagent and chromogen background, and include a peroxidase block in the chromogenic workflow (standard IHC practice). The selected example used 10% goat serum blocking, a peroxidase-conjugated anti-rabbit secondary for 30 minutes at 37°C, and DAB detection (datasheet A02018-1). If background persists, review blocking, washing and DAB development while keeping the EDTA pH 8.0 retrieval condition documented for this antibody (datasheet A02018-1; standard IHC practice). Judge improvements by clearer nuclei without loss of expected interphase nuclear staining (HPA: general nuclear expression; standard IHC practice).
How can I score NUMA1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and regions before scoring, then count interpretable nuclei while excluding damaged and necrotic areas (standard IHC scoring). Report the percentage of positive cells and, when intensity matters, an H-score from 0–300 based on the proportions at each intensity (standard IHC scoring). Normalise counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when area is the denominator (standard IHC scoring). Score mitotic cells separately when visible because NUMA1 moves from the interphase nucleus toward spindle poles and cortex during division (UniProt Q14980 localisation).
How do I distinguish genuine NUMA1 signal from artefact?
Look for staining in intact interphase nuclei across relevant cells, consistent with NUMA1’s nuclear-matrix residence and the HPA general nuclear pattern (UniProt Q14980 localisation; HPA: general nuclear expression). Evaluate apparent cytoplasmic or cortical positivity against mitotic morphology, because NUMA1 changes location during cell division (UniProt Q14980 localisation). Be cautious with colour limited to tissue edges, necrotic areas or nonspecific deposits, and check a section without primary antibody for detection background (standard IHC practice). A no-primary signal suggests background from the chromogenic workflow; confirm any proposed biological pattern in well-preserved interior regions using the same retrieval and development conditions (standard IHC practice).
Boster reagents
Best NUMA1 / Nuclear mitotic apparatus protein 1 IHC Antibodies
Both anti-NUMA1 antibodies have IHC images from human paraffin sections (catalog: A02018-1 and M02018-1 IHC captions); A02018-1 also has IF/ICC data from U2OS cells (catalog: A02018-1 IF caption).
Real IHC data
Anti-NUMA/NUMA1 Antibody ®
Cat # A02018-1
Real IHC data
Anti-NuMA NUMA1 Monoclonal Antibody
Cat # M02018-1
A02018-1 has IHC images from human prostate, breast, esophageal and lung cancer sections, plus an IF/ICC image from U2OS cells (catalog: A02018-1 image captions). M02018-1 has IHC images from human bladder and breast cancer sections and rat testis; IF/ICC is listed as an application, with no IF image supplied (catalog: M02018-1 image captions and applications).
Which to pick: For tissue IHC, choose M02018-1 if a rabbit monoclonal with human and rat paraffin-section examples suits the study; its pictured IHC used EDTA retrieval at pH 8.0 and 1:50 primary dilution (catalog: M02018-1 clone and IHC captions). Choose A02018-1 when a pictured IF/ICC result is useful: its U2OS image used 5 μg/ml, while its human paraffin-section IHC images used EDTA retrieval at pH 8.0 and 2 μg/ml (catalog: A02018-1 IF and IHC captions). For broader listed species reactivity, A02018-1 includes monkey alongside human, mouse and rat, while M02018-1 lists human, mouse and rat; fixative is unreported for the pictured paraffin sections of both antibodies (catalog: reactivity and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14980 (NUMA1_HUMAN, Nuclear mitotic apparatus protein 1).
- Human Protein Atlas. NUMA1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NUMA1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm..
- Human Protein Atlas. NUMA1 antibody validation summary (3 antibodies).
- Forty-five patient-derived xenografts capture the clinical and biological heterogeneity of Wilms tumor. Nature communications 2019 — PMC6925259.
- Morphologic and Immunohistochemical Characterization of Spontaneous Lymphoma/Leukemia in NSG Mice. Veterinary pathology 2020 — PMC8686194.
- LEF1 confers resistance to DNA-damaging chemotherapies through upregulation of PARP1 and NUMA1 in ovarian cancer. Oncogene 2025 — PMC12537492.
- Role of NuMA1 in breast cancer stem cells with implications for combination therapy of PIM1 and autophagy inhibition in triple negative breast cancer. Research square 2024 — PMC11030541.
- PubMed PMID:1541630 — UniProt-cited evidence.
- PubMed PMID:1541636 — UniProt-cited evidence.
- PubMed PMID:8408288 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUMA1 (Nuclear mitotic apparatus protein 1). Expected localisation Nuclear staining in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02018-1). Validated IHC antibody: A02018-1 at 1-2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human acinar adenocarcinoma of prostate tissue; fixative not specified (datasheet A02018-1); 2. Fixation: Image fixative and duration unreported (datasheet A02018-1); 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 A02018-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02018-1); 8. Primary antibody: Rabbit anti-NUMA1, 1-2 μg/ml (datasheet A02018-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02018-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02018-1); Caveat: Mitotic relocalization can affect nuclear scoring (UniProt); Isoform / epitope: 5 isoforms; confirm epitope coverage (UniProt); Regulation: Cell-cycle-dependent redistribution (UniProt). Evidence: UniProt Q14980, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm., Datasheet A02018-1, PMC6925259, PMC8686194. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUMA1/immunohistochemistry-ihc-guide.