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NUDT5 / ADP-sugar pyrophosphatase · IHC design guide
Design Immunohistochemistry for NUDT5
Plan paraffin-section NUDT5 IHC using the IHC-validated antibody and liver hepatocytes as a positive tissue reference (datasheet A07530; HPA tissue IHC). Score cytoplasmic staining while recording any nuclear signal separately, since the tissue profile and molecular annotation differ (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUDT5 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 in tissue; nucleus annotated molecularly (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining in several cell types, most abundant in liver (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07530) | |
| Positive control | Liver+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
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 A07530) | |
| Caveat | Nuclear annotation differs from observed tissue cytoplasm (UniProt; HPA tissue IHC) | |
| Regulation | Broadly expressed; most abundant in liver (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; one chain spanning residues 1–219 (UniProt) |
Section 1
Recommended NUDT5 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A07530) accompanies four published NUDT5 IHC protocols (PMC7258951; PMC12108576; PMC7877577; PMC10845800).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A07530) |
| Fixation | Image fixative and duration unreported (datasheet A07530); 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 A07530); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07530) |
| Primary antibody | Rabbit anti-NUDT5, 2-5 μg/ml (datasheet A07530) |
| Primary incubation | Overnight at 4 °C (datasheet A07530) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07530) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUDT5-positive staining in hepatocytes of liver (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several cell types, most abundant in liver. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet: A07530); the breast tissue microarray protocol used citrate pH 6.0 (PMC7877577).
Section 2
What Is the Expected NUDT5 Staining Pattern?
For paraffin-section IHC, expect cytoplasmic staining in hepatocytes and elongated or late spermatids, with medium staining reported in each (HPA tissue IHC). NUDT5 is also assigned to the nucleus and has no transmembrane segment (UniProt Q9UKK9). Interpret compartment differences cautiously: HPA tissue IHC is Approved, but reports low consistency between antibody staining and RNA expression; external characterization supports the antibody staining (HPA tissue IHC).
| Distinct cytoplasmic staining in hepatocytes, with a similar pattern in elongated or late spermatids. | This matches the reported medium-level positive cells (HPA tissue IHC). Compare signal with adjacent cells and the counterstain before scoring. A medium HPA category describes its reference observation; it does not prescribe a universal chromogen intensity or exposure setting (HPA tissue IHC; general IHC practice). |
| Strong staining confined to cell surfaces, extracellular material, or other unexpected structures. | Treat this as a localization warning because NUDT5 has no transmembrane segment and UniProt assigns it to the nucleus (UniProt Q9UKK9). Nuclear staining alone needs more careful review: it is biologically plausible, although the supplied tissue IHC profile emphasizes cytoplasm (UniProt Q9UKK9; HPA tissue IHC). |
| Strong, widespread staining in cells that HPA records as not detected, such as adipocytes. | Investigate cross-reactivity or endogenous detection activity before calling these cells positive (HPA tissue IHC; general IHC practice). HPA's 'not detected' is an assay observation, not proof that every cell lacks NUDT5; UniProt describes the protein as widely expressed (HPA tissue IHC; UniProt Q9UKK9). |
| Uniform chromogen haze across tissue compartments, including areas without clear cellular boundaries. | This does not establish NUDT5 localization. Review primary-antibody omission and detection controls, blocking, wash steps, and chromogen development as general IHC checks. Resolve the haze before comparing hepatocyte or spermatid staining with the HPA reference pattern (general IHC practice; HPA tissue IHC). |
| No discernible signal in hepatocytes of an otherwise interpretable liver section. | The result conflicts with the reported medium hepatocyte staining (HPA tissue IHC). Check that the IHC-validated antibody and detection system work on an appropriate control section, then review retrieval and antibody conditions. These are general assay checks; the supplied sources report no NUDT5-specific fixation sensitivity. |
💡Expected NUDT5 appearanceA convincing positive result shows cell-associated cytoplasmic staining in hepatocytes or elongated or late spermatids, near the reported medium category (HPA tissue IHC); diffuse haze or dominant surface or extracellular deposits should trigger control review (general IHC practice; UniProt Q9UKK9 topology).
| Tissue and cell selection | Liver hepatocytes and testicular elongated or late spermatids provide reported medium IHC examples (HPA tissue IHC). Kidney tubule cells, bronchial respiratory epithelial cells, skin fibroblasts, and tonsil squamous epithelial cells are reported low, so a faint result there is less decisive (HPA tissue IHC). |
| Compartment evidence | The tissue IHC summary describes cytoplasmic expression, whereas UniProt assigns NUDT5 to the nucleus (HPA tissue IHC; UniProt Q9UKK9). Score the compartment actually seen and disclose this difference; neither source makes every nuclear or cytoplasmic signal independently specific. |
| Antibody evidence | HPA019827 is Approved for IHC and ICC, while tissue IHC reliability is Approved with low antibody-staining/RNA consistency and supporting external characterization (HPA antibodies; HPA tissue IHC). Agreement with a reported pattern supports interpretation, but the supplied record does not establish IHC Enhanced validation. |
| Topology and processing | NUDT5 has no transmembrane segment, signal peptide, propeptide, or reported processed chain beyond residues 1–219 (UniProt Q9UKK9). These annotations give no basis to expect a membrane-bound or secreted IHC pattern; they also do not establish epitope accessibility after fixation. |
| IF/ICC Q&A: What localization should an IF image show? | HPA reports approved main localization in the nucleoplasm and vesicles, with additional centrosome and mid-piece localization (HPA subcellular ICC-IF). This is context for reading an IF result; the supplied evidence does not provide an IF protocol option for this IHC-P guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Liver control lacks hepatocyte staining. | Possible assay failure; HPA reports medium hepatocyte staining, but no NUDT5-specific fixation effect is supplied (HPA tissue IHC). | Confirm section integrity and detection controls, then review the IHC-validated antibody conditions and antigen retrieval as general IHC practice. Record the conditions used before comparing sections. |
| Chromogen appears broadly in negative-control tissue or after primary-antibody omission. | Endogenous detection activity or reagent background can produce signal without specific primary binding (general IHC practice). | Check the omission control and the detection chemistry; apply the appropriate endogenous-activity block for that chemistry, then compare with the same tissue stained using primary antibody (general IHC practice). |
| Strong signal appears in adipocytes recorded as not detected. | Cross-reactivity or assay background is possible; HPA's category reflects its observed assay result, not guaranteed molecular absence (HPA tissue IHC). | Compare cell boundaries and matched controls, then seek independent confirmation before scoring these cells as NUDT5-positive (general IHC practice). |
| Only surface or extracellular deposits dominate the section. | That distribution is poorly supported by the no-transmembrane topology and nuclear UniProt annotation (UniProt Q9UKK9). | Inspect the counterstain and omission control for deposits; compare with the HPA cytoplasmic IHC pattern and repeat under checked detection conditions if needed (HPA tissue IHC; general IHC practice). |
| Nuclear staining appears alongside, or instead of, cytoplasmic staining. | The sources differ by assay: UniProt assigns a nuclear location, HPA tissue IHC describes cytoplasm, and HPA ICC-IF identifies nucleoplasm (UniProt Q9UKK9; HPA tissue IHC; HPA subcellular ICC-IF). | Report nuclear and cytoplasmic scores separately, examine controls, and avoid treating the compartment alone as proof of specificity (general IHC practice). |
| Low-signal tissue seems negative while liver is positive. | Some cell types are reported low, including kidney tubule cells and bronchial respiratory epithelial cells, while hepatocytes are medium (HPA tissue IHC). | Use the liver result to verify the staining run, then document the detection threshold and score the low-signal tissue conservatively; absence of visible chromogen does not establish biological absence (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing NUDT5 staining
A07530 · IHCIHC analysis of NUDT5 using anti-NUDT5 antibody (A07530). NUDT5 was detected in a paraffin-embedded section of human liver cancer tissue. He
A07530 · IHCIHC analysis of NUDT5 using anti-NUDT5 antibody (A07530). NUDT5 was detected in a paraffin-embedded section of mouse liver tissue. Heat medi
A07530 · IFIF analysis of NUDT5 using anti-NUDT5 antibody (A07530) and anti-Beta Tubulin antibody (M01857-3). NUDT5 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000165609-NUDT5/tissue/cerebral+cortex#imid_5621316
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000165609-NUDT5/tissue/colon#imid_5621258
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000165609-NUDT5/tissue/liver#imid_5621243
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000165609-NUDT5/tissue/kidney#imid_5621244
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019827; green = NUDT5, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019827; green = NUDT5, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUDT5 IHC & IF
🧪Run liver first: hepatocytes should stain (HPA: Medium in liver hepatocytes). Use adipose tissue as the negative tissue: adipocytes should remain at background (HPA: Not detected in adipocytes); on the liver slide, score nonhepatocyte cells as internal negatives only if they are unstained, since the supplied HPA rows do not identify a negative liver cell type.
Positive control tissue: Liver (Hepatocytes, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUDT5 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Nucleoplasm (approved), Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control (caption: rabbit primary antibody), and NUDT5-knockout tissue as a biological negative. Quench endogenous peroxidase and check background in liver before interpreting HRP–DAB staining (caption: liver tissue and HRP–DAB detection).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported; the selected A07530 paraffin-section caption does not state a fixative. That caption reports heat-mediated retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required; the supplied evidence does not establish that frozen sections or IF/ICC are easier. For IF/ICC interpretation, HPA reports approved nucleoplasmic and vesicular localization (HPA: subcellular summary); liver endogenous peroxidase is a potential chromogenic background source (caption: liver tissue and HRP–DAB detection).
HPA tissue IHC evidence for NUDT5
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. External characterization data supports antibody staining.). 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 |
|---|---|---|---|---|
| Liver | Hepatocytes | Medium | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUDT5 IHC Tips
Troubleshoot NUDT5 chromogenic IHC by checking retrieval, cellular compartment, controls, and scoring before interpreting staining intensity.
What retrieval should I try first when NUDT5 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A07530). The selected paraffin-section example used that retrieval before 2 μg/ml primary antibody overnight at 4°C, so it provides a concrete starting condition for this antibody (datasheet A07530). If staining remains weak, compare a more or less intensive heating cycle on matched sections, keeping antibody concentration and detection conditions fixed (standard IHC practice). Include a section processed without primary antibody to identify detection background, and compare cellular staining with a liver control, where hepatocytes show medium staining (standard IHC practice; HPA: liver hepatocytes, medium).
Could fixation explain inconsistent NUDT5 staining across paraffin blocks?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report its fixative (datasheet A07530). Record each block’s fixative and fixation duration, then compare similarly processed sections before attributing an intensity difference to NUDT5 abundance (standard IHC practice). For a controlled comparison, use EDTA at pH 8.0, 2 μg/ml primary antibody overnight at 4°C, and the same chromogenic detection conditions across sections (datasheet A07530; standard IHC practice). Inspect morphology and staining together, and treat a block-specific loss of signal as a processing question until matched controls support a biological difference (standard IHC practice).
How should I assess nuclear and cytoplasmic NUDT5 staining?
Score nuclear and cytoplasmic staining separately because UniProt assigns NUDT5 to the nucleus, while HPA tissue IHC reports cytoplasmic expression in several cell types (UniProt Q9UKK9: subcellular location; HPA: tissue profile). HPA ICC/IF also reports nucleoplasm and vesicles as main locations, with centrosome and mid-piece staining as additional locations (HPA: subcellular). Use the counterstain and cell boundaries to assign each chromogenic signal to a compartment, and check matched sections for reproducibility (standard IHC practice). A cytoplasmic signal deserves evaluation against the tissue IHC pattern; nuclear staining alone should not be required for a positive call (HPA: tissue profile; UniProt Q9UKK9: subcellular location).
Can isoforms or epitope accessibility explain discordant NUDT5 staining?
The supplied record lists 0 isoforms and a 219-amino-acid chain, so an isoform switch is not supported as an explanation here (UniProt Q9UKK9: isoforms and processing). Its Nudix hydrolase domain spans residues 57–197, but the selected caption does not identify the antibody epitope (UniProt Q9UKK9: domains; datasheet A07530). Do not infer that a particular retrieval condition exposes a specific domain or modified residue without an epitope map (standard IHC practice). If staining differs across specimens, compare retrieval and processing on matched sections, then seek independent antibody or orthogonal evidence before assigning an epitope-based cause (standard IHC practice).
How can IF help resolve an ambiguous chromogenic NUDT5 pattern?
Use IF/ICC as a separate localisation check, rather than treating the selected paraffin-section IHC caption as an IF protocol (datasheet A07530). Multiplex NUDT5 with a marker identifying the cell population under study, and examine whether signal occupies the nucleoplasm or vesicles reported by HPA ICC/IF (HPA: subcellular; standard IF practice). Choose fluorophores after checking tissue autofluorescence and single-channel controls, so broad native fluorescence is not mistaken for NUDT5 (standard IF practice). Because NUDT5 has no transmembrane segment and its reported locations are intracellular, include a controlled permeabilisation step appropriate to the intracellular epitope, then compare it with a matched condition (UniProt Q9UKK9: topology; HPA: subcellular; standard IF practice).
How do I separate NUDT5 staining from chromogenic background?
The selected example used 10% goat serum blocking, a peroxidase-conjugated secondary antibody, and DAB development (datasheet A07530). Apply a peroxidase block and examine a no-primary control to reveal endogenous enzyme activity or detection-reagent background in the same tissue (standard chromogenic IHC practice). Compare diffuse haze, precipitate, and tissue-edge staining with intact cell-associated signal under the same development conditions (standard IHC practice). If background persists, titrate the primary around the documented 2 μg/ml condition while holding retrieval, blocking, detection, and exposure to DAB constant, then retain conditions that preserve interpretable cellular staining (datasheet A07530; standard IHC practice).
What should I quantify when NUDT5 staining varies between specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine whether the endpoint is a compartment-specific H-score, percentage of positive cells, or positive-cell density per mm², and apply one threshold across comparable sections (standard IHC practice). Record nuclear and cytoplasmic results separately because the supplied sources describe nuclear localisation and cytoplasmic tissue staining (UniProt Q9UKK9: subcellular location; HPA: tissue profile). Normalise counts to the number of evaluable cells or evaluable tissue area for the selected cell population, excluding folds, necrosis, and tissue edges (standard IHC practice). Keep retrieval, primary-antibody incubation, DAB development, imaging, and scoring rules consistent; include a reference section across staining runs (standard IHC practice).
Which NUDT5 patterns warrant a positive call rather than an artefact call?
A plausible positive pattern is reproducible cell-associated staining in a relevant compartment, supported by controls and morphology; liver hepatocytes provide a reported medium-staining reference (standard IHC practice; HPA: liver hepatocytes, medium). Evaluate nuclear signal against UniProt’s location and cytoplasmic signal against HPA tissue IHC, while recognising that these sources describe different assays (UniProt Q9UKK9: subcellular location; HPA: tissue profile). Distrust staining restricted to cut edges, necrotic regions, or cells with poor morphology, and check no-primary sections for endogenous peroxidase or detection background (standard IHC practice). HPA rates tissue staining Approved but reports low consistency with RNA data, so resolve disputed biological conclusions with independent evidence (HPA: tissue-IHC reliability).
Boster reagents
Best NUDT5 / ADP-sugar pyrophosphatase IHC Antibodies
A07530 has paraffin-section IHC images from human liver cancer and mouse and rat liver, plus IF/ICC data from U2OS cells (A07530 image captions). M07530 lists human, mouse and rat IHC reactivity (catalog).
Real IHC data
Anti-NUDT5 Antibody ®
Cat # A07530
A07530 has IHC images from paraffin sections of human liver cancer and mouse and rat liver, plus an IF/ICC image from U2OS cells (A07530 image captions). M07530 is listed for IHC in human, mouse and rat, with no IHC image supplied (catalog).
Which to pick: For tissue IHC, choose A07530 when documented paraffin-section staining matters: its captions specify EDTA retrieval at pH 8.0, but do not report the fixative (A07530 IHC captions). M07530 is a rabbit monoclonal, clone 26N82, listed for IHC without a supplied tissue image or processing details (catalog). For IF/ICC, choose A07530 based on its U2OS image at 5 μg/ml; for cross-species IHC, its images cover human, mouse and rat, while M07530 lists those species as reactive without supplied IHC images (A07530 image captions; catalog).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UKK9 (NUDT5_HUMAN, ADP-sugar pyrophosphatase).
- Human Protein Atlas. NUDT5 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NUDT5 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and vesicles. In addition localized to the centrosome and mid piece..
- Human Protein Atlas. NUDT5 antibody validation summary (1 antibodies).
- The high expression of MTH1 and NUDT5 predict a poor survival and are associated with malignancy of esophageal squamous cell carcinoma. PeerJ 2020 — PMC7258951.
- Assessment of NUDT5 in Endometrial Carcinoma: Functional Insights, Prognostic and Therapeutic Implications. Biomedicines 2025 — PMC12108576.
- The high expression of NUDT5 indicates poor prognosis of breast cancer by modulating AKT / Cyclin D signaling. PloS one 2021 — PMC7877577.
- The novel phosphatase NUDT5 is a critical regulator of triple-negative breast cancer growth. Breast cancer research : BCR 2024 — PMC10845800.
- PubMed PMID:10567213 — UniProt-cited evidence.
- PubMed PMID:10722730 — UniProt-cited evidence.
- PubMed PMID:11042152 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUDT5 (ADP-sugar pyrophosphatase). Expected localisation Cytoplasmic in tissue; nucleus annotated molecularly (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07530). Validated IHC antibody: A07530 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Liver (Hepatocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A07530); 2. Fixation: Image fixative and duration unreported (datasheet A07530); 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 A07530); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07530); 8. Primary antibody: Rabbit anti-NUDT5, 2-5 μg/ml (datasheet A07530); 9. Primary incubation: Overnight at 4 °C (datasheet A07530). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07530); Caveat: Nuclear annotation differs from observed tissue cytoplasm (UniProt; HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; one chain spanning residues 1–219 (UniProt); Regulation: Broadly expressed; most abundant in liver (UniProt). Evidence: UniProt Q9UKK9, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and vesicles. In addition…, Datasheet A07530, PMC7258951, PMC12108576. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUDT5/immunohistochemistry-ihc-guide.