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NUDT21 / Cleavage and polyadenylation specificity factor subunit 5 · IHC design guide
Design Immunohistochemistry for NUDT21
Plan NUDT21 paraffin-section IHC around the reported cytoplasmic tissue pattern, with high staining in kidney tubule cells and parathyroid and stomach glandular cells (HPA tissue IHC). The catalog antibody lists an IHC dilution range of 1:50–1:200 (datasheet A09247); score intensity and compartment against matched controls (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUDT21 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 tissue staining (HPA tissue IHC); nuclear location (UniProt) | |
| Staining pattern | Broad cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A09247) | |
| Positive control | Kidney+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 fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining reliability is uncertain; verify locally (HPA tissue IHC) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | 0 isoforms; no extracellular epitope domain (UniProt) |
Section 1
Recommended NUDT21 IHC & IF Protocols
Start with the catalog antibody’s IHC-P protocol (datasheet A09247), then compare the published breast tissue protocol (PMC7200255).
| Sample | Paraffin-embedded human esophageal cancer tissue; fixative not specified (datasheet A09247) |
| Fixation | Image fixative and duration unreported (datasheet A09247); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A09247); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-NUDT21, 1:50-1:200 (datasheet A09247) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUDT21-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteTry heat-mediated citrate retrieval at pH 6.0 first (datasheet A09247); the breast tissue study also used boiled citrate buffer (PMC7200255).
Section 2
What Is the Expected NUDT21 Staining Pattern?
NUDT21 is found in the nucleus and cytoplasm and has no transmembrane segment (UniProt O43809). In paraffin-section IHC, HPA describes a ubiquitous cytoplasmic pattern, with high staining in kidney tubule cells and parathyroid and stomach glandular cells (HPA tissue IHC). Interpret that pattern cautiously: HPA rates its tissue IHC reliability uncertain, citing medium agreement with RNA expression and pending external verification (HPA tissue IHC).
| Cell-associated cytoplasmic staining in kidney tubules or parathyroid or stomach glands, with clear tissue structure. | This matches HPA's high cell-specific IHC observations and its broad cytoplasmic profile (HPA tissue IHC). Compare intensity within a run rather than treating every cell as equally positive: HPA also reports medium, low and undetected cell populations (HPA tissue IHC). |
| Strong staining confined to cell membranes, luminal deposits or extracellular material. | This compartment does not match UniProt's nucleus/cytoplasm assignment or its lack of a transmembrane segment (UniProt O43809 topology). Treat it as a possible staining artefact and examine morphology and controls before scoring; nuclear staining alone is not a mismatch because nuclear localization is reported (UniProt O43809). |
| Strong signal in adipocytes or ovarian stromal cells, while expected positive cells are weak. | HPA reports NUDT21 as not detected in those cell populations (HPA tissue IHC). Unexpected staining raises cross-reactivity or endogenous detection activity as possibilities (standard IHC practice); it does not prove either, especially given HPA's uncertain tissue IHC reliability (HPA tissue IHC). |
| Brown signal spreads across stroma, empty spaces and many cell types without crisp cell boundaries. | Diffuse, noncellular signal is more consistent with background than a scorable intracellular pattern (standard chromogenic IHC practice; UniProt O43809 localization). Check a no-primary control and assess blocking, washing and detection conditions (standard IHC practice). HPA's broad expression alone cannot establish that diffuse staining is specific (HPA tissue IHC). |
| No signal in kidney tubule cells or parathyroid or stomach glandular cells. | These are reported high IHC populations, so an absent result warrants a run-level check (HPA tissue IHC). Confirm tissue preservation and the assay's positive control, then review antibody dilution, retrieval and detection as general IHC variables (standard IHC practice). HPA provides no NUDT21-specific retrieval or fixation rule. |
💡Expected NUDT21 appearanceA plausible positive result is cell-associated cytoplasmic staining reaching high intensity in kidney tubule cells or parathyroid or stomach glandular cells (HPA tissue IHC); membrane-only or extracellular deposits are suspect against the reported intracellular localization (UniProt O43809).
| Compartment reported by each method | HPA tissue IHC describes ubiquitous cytoplasmic expression, whereas UniProt lists nucleus and cytoplasm and describes shuttling (HPA tissue IHC; UniProt O43809). Score cytoplasmic IHC against the tissue profile without declaring nuclear signal biologically impossible. |
| IF/ICC Q: where should puncta appear? | A: HPA supports nuclear bodies as the main IF/ICC location; its additional centriolar-satellite assignment is uncertain (HPA subcellular ICC-IF). UniProt reports punctate subnuclear paraspeckles (UniProt O43809). Do not require individually resolved puncta in chromogenic tissue IHC. |
| Tissue and cell selection | Kidney tubules, parathyroid glands and stomach glands are reported high; adipocytes and ovarian stromal cells are reported not detected (HPA tissue IHC). Use cell identity within the section when assessing a result, since HPA's overall profile is broad (HPA tissue IHC). |
| Antibody-level validation | HPA019863 has an uncertain IHC assessment, while HPA074228 has supported ICC assessment and no listed IHC assessment (HPA antibodies). An IF validation label should not be carried over to paraffin-section IHC. |
| Topology and processing | UniProt reports no transmembrane segment or signal peptide and lists a chain spanning residues 2–227 (UniProt O43809 topology/processing). That record supports an intracellular expectation; it supplies no basis for a shed or secreted staining pattern. |
| Fixation sensitivity | A target-specific fixation effect is unreported in the supplied UniProt and HPA records. Assess retrieval and tissue processing through ordinary IHC controls (standard IHC practice); do not infer a NUDT21-specific fixation response from topology, modifications or tissue intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank across the run. | A staining-run failure is possible; HPA reports high signal in kidney tubules and parathyroid and stomach glandular cells (HPA tissue IHC). | Check an appropriate positive control, tissue integrity, primary-antibody application and detection reagents; review retrieval and dilution as general IHC variables (standard IHC practice). |
| Positive cells stain weakly but background is low. | The assay may be underdeveloped, or the sampled population may normally stain less strongly: HPA records medium and low populations alongside high ones (HPA tissue IHC). | Confirm the cell type first, then compare a reported high population in the same run; adjust detection or antibody conditions only against controls (HPA tissue IHC; standard IHC practice). |
| Diffuse brown haze obscures cell boundaries. | Nonspecific antibody binding, insufficient washing or endogenous detection activity can produce background (standard chromogenic IHC practice). | Inspect no-primary and detection controls; review blocking and washes before interpreting intensity (standard IHC practice). Do not score haze as HPA's cytoplasmic pattern (HPA tissue IHC). |
| Signal is concentrated at membranes or outside cells. | The location conflicts with reported nuclear/cytoplasmic localization and absence of a transmembrane segment (UniProt O43809). | Check morphology and controls, and compare another section or validated antibody when available (standard IHC practice). Record the compartment discrepancy before calling the section NUDT21-positive. |
| A reported negative population stains as strongly as the positive control. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice); HPA's negative-cell calls carry the same uncertain tissue IHC reliability caveat (HPA tissue IHC). | Verify cell identity and no-primary control, then compare antibody-independent controls if available (standard IHC practice). Report the discordance rather than treating one HPA call as definitive. |
| IF/ICC shows nuclear puncta but tissue IHC looks mostly cytoplasmic. | The methods have different reported observations: HPA supports nuclear bodies in ICC-IF and describes cytoplasmic tissue IHC (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret each image against its own method and antibody validation status; do not use puncta as a required chromogenic IHC scoring feature (HPA antibodies; standard IHC practice). |
Experiment evidence
IHC & IF images showing NUDT21 staining
A09247 · IHCImmunohistochemistry of paraffin-embedded human esophageal cancer using NUDT21 Rabbit pAb at dilution of 1:100 .Perform high pressure antige
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000167005-NUDT21/tissue/cerebral+cortex#imid_13820953
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000167005-NUDT21/tissue/colon#imid_13820879
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000167005-NUDT21/tissue/liver#imid_13820888
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000167005-NUDT21/tissue/kidney#imid_13820889
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA074228; green = NUDT21, blue = nucleus, red = microtubules.
HPA · ICC-IF PC-3Human Protein Atlas immunofluorescence, PC-3, antibody HPA074228; green = NUDT21, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUDT21 IHC & IF
🧪Run kidney first: tubular cells should stain (HPA: High in kidney tubular cells). Use adipose tissue as the negative tissue (HPA: adipocytes Not detected); cells without specific signal on the kidney slide should show counterstain without convincing nuclear chromogen, but the supplied HPA rows do not establish a kidney cell type as a true internal negative.
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUDT21 in PC-3, Rh30, U2OS, with annotated localisation: Nuclear bodies (supported) (HPA subcellular).
Technical controls: Run a no-primary (secondary-only) control, a concentration-matched nonimmune rabbit polyclonal IgG control, and a verified NUDT21 knockout preparation as a biological negative (A09247 caption: rabbit pAb; standard IHC practice). For kidney IHC, quench endogenous peroxidase and check endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative in the selected A09247 paraffin-section caption is unreported (A09247 caption). That example uses high-pressure retrieval in 10 mM citrate, pH 6.0, with antibody at 1:100; it does not establish that retrieval is required for every specimen (A09247 caption). No matched evidence establishes that frozen sections or IF are easier; kidney background from endogenous peroxidase or biotin can complicate chromogenic detection (standard IHC practice).
HPA tissue IHC evidence for NUDT21
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUDT21 IHC Tips
Troubleshoot NUDT21 staining in paraffin sections by checking retrieval, compartment, tissue context and controls before comparing chromogenic scores.
Which retrieval conditions should I try first for weak NUDT21 staining?
Start with heat-mediated retrieval in 10 mM citrate buffer at pH 6.0 (datasheet A09247). The selected paraffin-section image used high-pressure retrieval in that buffer and a 1:100 primary dilution, providing a reference combination for this antibody (A09247 tissue-IHC caption). Keep heating and cooling conditions consistent across slides, then compare signal with a matched section processed without primary antibody. If staining remains weak, adjust retrieval time in small steps while watching for tissue damage; document each change before altering primary-antibody concentration. Judge improvement by cellular staining and preserved morphology, rather than a darker background alone.
How should I troubleshoot suspected fixation effects on NUDT21 IHC?
The selected image documents paraffin-embedded tissue but does not state its fixative, so target-specific fixation sensitivity is unknown (A09247 tissue-IHC caption). Record the actual fixative and fixation duration for each specimen before attributing weak staining to fixation. Compare sections from specimens with known, similar handling under the same 10 mM citrate, pH 6.0 retrieval and detection conditions (datasheet A09247). Check morphology and a control stain on adjacent sections when staining varies between blocks. Treat any benefit from changing retrieval or antibody incubation as an empirical result for those specimens, not evidence that NUDT21 has a particular fixation requirement.
Should NUDT21 appear in nuclei, cytoplasm, or both?
Evaluate nuclear and cytoplasmic staining separately: NUDT21 is annotated in both compartments and can shuttle between them (UniProt O43809). Punctate subnuclear structures adjacent to nuclear speckles are also reported (UniProt O43809), while nuclear bodies are the supported main IF location (HPA subcellular). HPA tissue IHC describes ubiquitous cytoplasmic expression, but assigns the tissue staining uncertain reliability (HPA tissue IHC). Use a nuclear counterstain and matched negative control to decide whether apparent nuclear puncta lie within intact cells. If a section shows only diffuse extracellular color, review morphology, retrieval and detection background before scoring it as NUDT21.
Could an isoform or modified epitope explain inconsistent staining?
The supplied record lists 0 isoforms, so an annotated isoform switch does not explain different staining patterns here (UniProt O43809). NUDT21 has a Nudix domain at residues 76–201 and several reported modified residues near its N terminus (UniProt O43809). Without a mapped epitope for the catalog antibody, do not assign loss of staining to one modification or domain. Compare matched sections using the same retrieval and primary-antibody conditions, then check whether any difference persists across independent specimens. If epitope access remains a concern, document the antibody's stated immunogen or epitope before testing alternative conditions or an independently validated antibody.
How can IF help resolve ambiguous chromogenic NUDT21 staining?
Use IF as a separate validation experiment, since the selected antibody evidence here is a paraffin-section chromogenic image (A09247 tissue-IHC caption). Multiplex NUDT21 with a validated marker for the expected cell type and a nuclear counterstain, then inspect single channels before overlays. Select a fluorophore in a channel with low autofluorescence for the specimen, and include unstained and secondary-only controls. Because NUDT21 is intracellular and has no transmembrane segment, choose permeabilisation that permits access to nuclear and cytoplasmic epitopes; verify the antibody's epitope location if known (UniProt O43809). Interpret nuclear puncta against the reported nuclear-body location, while treating centriolar-satellite localisation as uncertain (HPA subcellular).
What should I check when NUDT21 staining is diffusely brown?
Compare a no-primary section with the stained section to identify color produced by the detection workflow rather than primary-antibody binding. Apply a peroxidase block before chromogenic detection and monitor DAB development under the same conditions for every section; these are general IHC workflow steps. If background persists, review blocking, washes, primary-antibody concentration and retrieval intensity one variable at a time. The selected paraffin-section image used 1:100 primary antibody after 10 mM citrate retrieval at pH 6.0 (A09247 tissue-IHC caption). Require staining to follow intact cellular compartments before counting it, especially because HPA rates tissue-IHC reliability uncertain (HPA tissue IHC).
How should I score NUDT21 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Choose a predefined compartment and score nuclear and cytoplasmic staining separately, because both locations are annotated for NUDT21 (UniProt O43809). For each defined cell population, record the percentage of positive cells and an intensity category, then calculate an H-score if intensity is reproducible. Alternatively, report positive-cell density per mm² of viable tissue when cell counts or tissue area vary substantially. Normalise comparisons to the number of eligible cells or viable tissue area, and apply identical thresholds, retrieval and DAB development across sections. Report the cell population and compartment with each score; HPA tissue-IHC reliability is uncertain, so staining intensity alone has limited interpretive support (HPA tissue IHC).
How do I distinguish genuine NUDT21 staining from tissue artefact?
Look for staining within intact nuclei or cytoplasm, the annotated NUDT21 compartments, and assess nuclear puncta in light of its reported subnuclear localisation (UniProt O43809). Check whether the labeled cell population fits the specimen: HPA reports high staining in kidney tubular cells and stomach glandular cells, while its tissue-IHC reliability is uncertain (HPA tissue IHC). Exclude section edges, folds, necrotic regions and extracellular deposits from scoring when morphology or no-primary controls indicate artefact. A positive no-primary control can implicate endogenous enzyme activity or detection chemistry; review the peroxidase block and DAB conditions. Confirm unexpected compartment or cell-type patterns with an independent validation method before assigning biological meaning.
Boster reagents
Best NUDT21 / Cleavage and polyadenylation specificity factor subunit 5 IHC Antibodies
The IHC-validated antibody has an image from a human esophageal cancer paraffin section; the catalog also lists human, mouse, and rat reactivity and IF/ICC applications for another SKU (catalog applications; A09247 IHC image caption).
Real IHC data
Anti-NUDT21 Antibody
Cat # A09247
A09247 will render with its IHC image from a human esophageal cancer paraffin section (A09247 IHC image caption). A09247-1 is listed for IHC and IF/ICC in human, mouse, and rat, but has no supplied IHC or IF image and will not render as a card (catalog applications, reactivity, and image captions).
Which to pick: Choose A09247 for paraffin-section tissue IHC because its own image shows that use with citrate pH 6.0 retrieval; the fixative is unreported (A09247 IHC image caption). For IF/ICC, consider A09247-1 because those applications are listed, although no IF image is supplied (catalog applications and image captions). Both rabbit polyclonal antibodies list human, mouse, and rat reactivity, but the supplied tissue image documents human IHC only (catalog host, clone, and reactivity; A09247 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43809 (CPSF5_HUMAN, Cleavage and polyadenylation specificity factor subunit 5).
- Human Protein Atlas. NUDT21 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. NUDT21 subcellular location (ICC-IF): Mainly localized to the nuclear bodies. In addition localized to the centriolar satellites..
- Human Protein Atlas. NUDT21 antibody validation summary (2 antibodies).
- NUDT21 Suppresses Breast Cancer Tumorigenesis Through Regulating CPSF6 Expression. Cancer management and research 2020 — PMC7200255.
- Silencing NUDT21 Attenuates the Mesenchymal Identity of Glioblastoma Cells via the NF-κB Pathway. Frontiers in molecular neuroscience 2017 — PMC5742174.
- Pan-cancer analysis of NUDT21 and its effect on the proliferation of human head and neck squamous cell carcinoma. Aging 2024 — PMC10929839.
- A Biomimetic Dual-Targeting Nano-APA-Editor Reprograms the 3'UTR Landscape for Tongue Squamous Cell Carcinoma Therapy. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13336823.
- PubMed PMID:9659921 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUDT21 (Cleavage and polyadenylation specificity factor subunit 5). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A09247). Validated IHC antibody: A09247 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human esophageal cancer tissue; fixative not specified (datasheet A09247); 2. Fixation: Image fixative and duration unreported (datasheet A09247); 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: Citrate pH 6.0 (datasheet A09247); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-NUDT21, 1:50-1:200 (datasheet A09247); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Staining reliability is uncertain; verify locally (HPA tissue IHC); Isoform / epitope: 0 isoforms; no extracellular epitope domain (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). Evidence: UniProt O43809, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the nuclear bodies. In addition localized to the…, Datasheet A09247, PMC7200255. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUDT21/immunohistochemistry-ihc-guide.