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NUBPL / Iron-sulfur cluster transfer protein NUBPL · IHC design guide
Design Immunohistochemistry for NUBPL
Plan chromogenic paraffin IHC for NUBPL using cytoplasmic staining as the tissue readout (HPA tissue IHC); NUBPL is a mitochondrial protein (UniProt). Start with the 2–5 μg/ml IHC antibody range (datasheet A10634-1), and interpret staining cautiously because antibody staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NUBPL 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) | |
| Staining pattern | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A10634-1) | |
| Positive control | Fallopian tube+4 more · see all | |
| Negative control | Caudate+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended NUBPL IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A10634-1) sits alongside published NUBPL staining of colorectal carcinoma (PMC5480060), bladder cancer (PMC11407088), and myocardium (PMC11897062).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A10634-1) |
| Fixation | Image fixative and duration unreported (datasheet A10634-1); 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 A10634-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A10634-1) |
| Primary antibody | Rabbit anti-NUBPL, 2-5 μg/ml (datasheet A10634-1) |
| Primary incubation | Overnight at 4 °C (datasheet A10634-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A10634-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NUBPL-positive staining in ciliated cells (tip of cilia) of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A10634-1); the bladder study reports retrieval without specifying a buffer (PMC11407088).
Section 2
What Is the Expected NUBPL Staining Pattern?
NUBPL localizes to mitochondria (UniProt Q8TB37; HPA subcellular ICC-IF: supported). In paraffin-section IHC, expect cytoplasmic staining in many tissues, including glandular cells in the colon and duodenum (HPA tissue IHC: medium). The protein has no transmembrane segment (UniProt Q8TB37 topology). Treat intensity as a guide: HPA rates tissue IHC reliability Approved but reports low consistency with RNA expression and pending external verification (HPA tissue IHC).
| Cytoplasmic staining in colon or duodenal glandular cells, with little staining in adjacent negative-control material. | This matches the reported medium glandular-cell signal (HPA tissue IHC: colon and duodenum). Mitochondrial localization supports a cytoplasmic distribution (UniProt Q8TB37; HPA subcellular ICC-IF), but chromogenic IHC need not resolve individual mitochondria (general IHC practice). |
| Predominantly nuclear, cell-surface, or extracellular deposit with little cytoplasmic signal. | That distribution conflicts with mitochondrial localization (UniProt Q8TB37; HPA subcellular ICC-IF). Check morphology, counterstain and detection controls before assigning it to NUBPL (general IHC practice); compartment alone cannot identify the artefact's cause. |
| Strong staining in cells listed as undetected, such as prostate glandular cells or lymph-node germinal-center cells. | Those cell-level findings differ from HPA observations (HPA tissue IHC: not detected). Consider cross-reactivity or endogenous detection activity, then compare controls and an independently validated antibody (general IHC practice). An HPA negative is an observed result, not proof that every specimen must be negative. |
| A broad, even deposit obscures cell boundaries across positive and negative regions. | Diffuse background limits interpretation of the reported cytoplasmic pattern (HPA tissue IHC: cytoplasmic in most tissues). Review no-primary and detection controls, blocking and wash conditions (general IHC practice); background cannot establish NUBPL expression. |
| No convincing signal in glandular cells of an otherwise interpretable colon or duodenum section. | These are useful expected-positive cell populations because HPA reports medium staining (HPA tissue IHC: colon and duodenum). Recheck antibody performance and tissue integrity with controls (general IHC practice); absent staining alone does not resolve whether expression or assay performance differs. |
💡Expected NUBPL appearanceCall the result positive when cytoplasmic staining is discernible in colon or duodenal glandular cells at roughly medium intensity (HPA tissue IHC); dominant nuclear or extracellular deposit argues against the supported mitochondrial localization (UniProt Q8TB37; HPA subcellular ICC-IF).
| Compartment resolution | NUBPL is mitochondrial and lacks a transmembrane segment (UniProt Q8TB37; HPA subcellular ICC-IF). Score a cytoplasmic IHC pattern without requiring visible mitochondrial puncta (general IHC practice). |
| Tissue and cell selection | HPA reports high staining at fallopian-tube cilia tips, medium staining in several glandular epithelia, and no detection in selected cell populations (HPA tissue IHC). Compare the named cell type, not whole-tissue averages. |
| Isoforms and processing | UniProt lists two isoforms and a 39–319 mature chain (UniProt Q8TB37). These annotations alone do not identify the antibody epitope or predict different IHC staining between isoforms. |
| Strength of validation | HPA lists HPA029203 as IHC Approved, yet tissue staining has low consistency with RNA expression and awaits external verification (HPA antibodies; HPA tissue IHC). Interpret unexpected patterns with controls and independent evidence. |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon or duodenal glandular cells show no signal. | Assay failure or specimen variation is possible; HPA reports medium staining in these cells (HPA tissue IHC). | Check a known-performing positive section, no-primary control, antigen-retrieval record and detection steps (general IHC practice); do not infer a NUBPL-specific retrieval requirement. |
| Nuclear staining dominates the section. | The compartment differs from supported mitochondrial localization (UniProt Q8TB37; HPA subcellular ICC-IF); the cause is unresolved. | Compare no-primary and secondary-detection controls, then assess whether cytoplasmic staining remains (general IHC practice). |
| Prostate glandular cells stain strongly. | HPA reports these cells as not detected (HPA tissue IHC); cross-reactivity or detection background is possible. | Inspect matched controls and repeat with an independently validated antibody if the finding is central (general IHC practice). |
| Diffuse chromogen covers several cell types. | Background can obscure the reported cytoplasmic pattern (HPA tissue IHC: cytoplasmic in most tissues). | Inspect no-primary controls and optimize blocking, antibody concentration and washes using standard IHC controls (general IHC practice). |
| Fallopian-tube signal appears only at cilia tips. | That location and high staining level match the reported ciliated-cell observation (HPA tissue IHC: fallopian tube). | Record the exact cell compartment and compare neighboring structures; confirm an unexpected distribution with independent evidence (general IHC practice). |
| Q: Should IF/ICC show mitochondrial signal? | A: HPA reports supported mitochondrial localization in ICC-IF, with images from A-431, Hep-G2 and U2OS (HPA subcellular ICC-IF). | Use the separate IF/ICC guide for assay design; compare any IF result with a mitochondrial reference and controls (general IF practice). |
Experiment evidence
IHC & IF images showing NUBPL staining
A10634-1 · IHCIHC analysis of NUBPL using anti-NUBPL antibody (A10634-1). NUBPL was detected in a paraffin-embedded section of human liver cancer tissue.
A10634-1 · IHCIHC analysis of NUBPL using anti-NUBPL antibody (A10634-1). NUBPL was detected in a paraffin-embedded section of human lung cancer tissue. H
A10634-1 · IFIF analysis of NUBPL using anti-NUBPL antibody (A10634-1). NUBPL was detected in an immunocytochemical section of HELA cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151413-NUBPL/tissue/cerebral+cortex#imid_7293664
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151413-NUBPL/tissue/colon#imid_7289455
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151413-NUBPL/tissue/liver#imid_7289456
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151413-NUBPL/tissue/kidney#imid_7289449
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029203; green = NUBPL, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029203; green = NUBPL, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NUBPL IHC & IF
🧪Run fallopian tube first; its ciliated cells, particularly the tips of cilia, should stain (HPA: High in fallopian tube ciliated cells). Use caudate glial cells as a negative tissue (HPA: Not detected in caudate glial cells), and expect background-level signal in areas of the positive slide outside the reported ciliated-cell staining pattern rather than assuming every neighboring cell is NUBPL-negative.
Positive control tissue: Fallopian tube (Ciliated cells (tip of cilia), HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NUBPL in A-431, Hep-G2, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a nonimmune rabbit IgG isotype control matched to the primary antibody’s class and clonality where known (selected-SKU caption: rabbit anti-NUBPL); use NUBPL knockout material, or an immunizing-peptide block if the peptide is available, as a biological specificity control. Quench endogenous peroxidase for chromogenic detection and check for nonspecific signal along ciliary edges (HPA: High at tips of fallopian tube cilia).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU tissue-IHC caption does not state a fixative. The reported paraffin-section example used heat retrieval in EDTA at pH 8.0 (selected-SKU tissue-IHC caption); retrieval dependence in fallopian tube has not been established by that example. Neither frozen sections nor IF is shown to be easier by the supplied evidence; ciliary-tip staining warrants careful background assessment because the expected subcellular location is mitochondria (HPA: High at tips of fallopian tube cilia; HPA subcellular: mitochondria supported).
HPA tissue IHC evidence for NUBPL
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Fallopian tube | Ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NUBPL IHC Tips
These questions address NUBPL staining and interpretation in paraffin sections with chromogenic IHC; the immunofluorescence entry covers a separate application.
Which retrieval condition should I start with for NUBPL in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin-section NUBPL IHC (datasheet A10634-1). This condition was used for the selected human liver cancer image, with 2 μg/ml primary antibody incubated overnight at 4°C (caption A10634-1). If staining is weak, compare retrieval times on adjacent sections while keeping antibody concentration and DAB development constant (standard IHC practice). Run a positive reference section and a no-primary control alongside them to judge signal and background (standard IHC practice). The caption supports this retrieval condition for that image, but does not establish an optimal heating time across specimens (caption A10634-1).
How should I assess whether fixation is limiting NUBPL staining?
The selected caption identifies a paraffin section but does not state its fixative, so NUBPL-specific fixation sensitivity is unknown (caption A10634-1). Record the fixative and fixation duration for each specimen before comparing staining results (standard IHC practice). If specimens vary, stain sections together using EDTA at pH 8.0 and the same antibody incubation to reduce processing differences (datasheet A10634-1; standard IHC practice). Compare tissue morphology, the expected intracellular pattern, and a reference section before attributing weak signal to fixation (UniProt Q8TB37; standard IHC practice). Neither the reported tissue distribution nor NUBPL topology establishes how fixation affects this antibody (HPA tissue IHC; UniProt Q8TB37).
What intracellular pattern supports specific NUBPL staining in IHC?
Expect NUBPL signal primarily in the mitochondrial compartment, interpreted within the cytoplasm of intact cells (UniProt Q8TB37; HPA subcellular). At light-microscope resolution, a granular cytoplasmic pattern is compatible with that location, but DAB alone cannot prove mitochondrial colocalisation (standard IHC practice). HPA describes cytoplasmic expression in most tissues and reports a high signal at cilia tips in fallopian-tube ciliated cells (HPA tissue IHC). Check cell identity and morphology before treating that unusual pattern as a general NUBPL localisation rule (HPA tissue IHC). Strong nuclear-only or diffuse extracellular staining warrants review of controls, retrieval, and background before scoring (UniProt Q8TB37; standard IHC practice).
Can this IHC antibody distinguish NUBPL isoforms or processed protein?
NUBPL has 2 annotated isoforms and a reported mature chain spanning residues 39–319 (UniProt Q8TB37). The supplied material does not map this antibody’s epitope, so staining cannot establish which isoform or processing state it detects (caption A10634-1; UniProt Q8TB37). Because no transmembrane segment is annotated, do not infer an antibody-accessible membrane face from topology alone (UniProt Q8TB37). If isoform specificity matters, obtain an epitope map and compare it with isoform sequences before interpreting section staining (standard IHC practice). Use an orthogonal, isoform-resolving assay where that distinction is essential; DAB intensity alone cannot make it (standard IHC practice).
How should I investigate NUBPL localisation with multiplex immunofluorescence?
Treat IF/ICC as a separate assay: HPA reports supported mitochondrial localisation and lists A-431, Hep-G2, and U2OS among cell lines with ICC/IF images (HPA subcellular). Pair NUBPL with a marker of the expected cell type and a mitochondrial marker when testing whether signal lies in the predicted compartment (UniProt Q8TB37; standard IF practice). Choose fluorophores after checking tissue autofluorescence, and include single-label controls to assess bleed-through (standard IF practice). Permeabilise sufficiently to expose the antibody’s epitope, but establish the condition empirically because its location relative to mitochondrial membranes is not supplied (UniProt Q8TB37; standard IF practice). The paraffin-section caption provides no IF fixation or permeabilisation condition (caption A10634-1).
How can I separate NUBPL DAB signal from tissue background?
Start with a no-primary control and inspect unstained tissue pigment before assigning brown deposits to NUBPL (standard IHC practice). Block endogenous peroxidase for HRP/DAB detection, then assess whether any residual colour follows tissue structures or damaged areas (standard IHC practice). The selected image used 10% goat serum before 2 μg/ml primary antibody and an HRP/DAB workflow (caption A10634-1). If background remains high, compare primary-antibody concentrations and blocking conditions on matched sections while keeping DAB development consistent (standard IHC practice). Prioritise intracellular staining with plausible mitochondrial distribution over diffuse extracellular deposits or section-edge staining (UniProt Q8TB37; standard IHC practice).
What should I measure when comparing NUBPL staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score NUBPL in a predefined cell population and anatomical region, recording both the percentage of positive cells and staining intensity (standard IHC practice). An H-score sums the percentage at each intensity level multiplied by its 0–3 intensity score, yielding 0–300 (standard IHC practice). Alternatively, report positive-cell density per mm² when cell counts and tissue area are reliable (standard IHC practice). Normalise to the number of eligible cells or analyzable tissue area, and apply the same threshold and DAB development across sections (standard IHC practice). Report compartments separately if mixed cell populations or necrosis would distort the comparison (standard IHC practice).
When is an apparent NUBPL-positive area likely to be artefactual?
A convincing NUBPL result should occur within identifiable cells and be compatible with its supported mitochondrial localisation (UniProt Q8TB37; HPA subcellular). Reconsider nuclear-only, extracellular, section-edge, or necrotic staining, especially when comparable deposits occur in the no-primary control (UniProt Q8TB37; standard IHC practice). Check whether the stained cell type fits the section: HPA reports medium signal in colon glandular cells but no detected signal in lymph-node germinal-center cells (HPA tissue IHC). Interpret those comparisons cautiously because HPA marks tissue staining Approved while reporting low agreement with RNA expression and pending external verification (HPA tissue IHC). Residual endogenous peroxidase can also mimic DAB positivity, so verify the peroxidase block (standard IHC practice).
Boster reagents
Best NUBPL / Iron-sulfur cluster transfer protein NUBPL IHC Antibodies
A10634-1 has IHC images from paraffin-embedded human liver cancer, lung cancer and placenta sections, and an IF image from HeLa cells (catalog image captions). Human and mouse reactivity are listed (catalog reactivity).
Real IHC data
Anti-NUBPL Antibody ®
Cat # A10634-1
A10634-1 will render with its own human liver cancer paraffin-section IHC figure; additional IHC captions describe human lung cancer and placenta sections (catalog IHC image captions). The same SKU lists IHC, ICC and IF applications and has an IF caption for HeLa cells (catalog applications; catalog IF image caption).
Which to pick: Choose A10634-1 for human paraffin-section IHC: its IHC captions document that preparation, while the fixative is unreported (catalog IHC image captions). Choose A10634-1 for IF/ICC based on its listed applications and HeLa cell IF image; it is rabbit-derived, but clonality is unreported (catalog applications; catalog IF image caption; catalog host and clone fields). For mouse samples, A10634-1 lists mouse reactivity, but its supplied IHC and IF captions document human samples only (catalog reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TB37 (NUBPL_HUMAN, Iron-sulfur cluster transfer protein NUBPL).
- Human Protein Atlas. NUBPL tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NUBPL subcellular location (ICC-IF): Mainly localized to the mitochondria..
- Human Protein Atlas. NUBPL antibody validation summary (1 antibodies).
- NUBPL, a novel metastasis-related gene, promotes colorectal carcinoma cell motility by inducing epithelial-mesenchymal transition. Cancer science 2017 — PMC5480060.
- Disulfidptosis and ferroptosis related genes define the immune microenvironment and NUBPL serves as a potential biomarker for predicting prognosis and immunotherapy response in bladder cancer. Heliyon 2024 — PMC11407088.
- Early embryonic lethality in complex I associated p.L104P Nubpl mutant mice. Orphanet journal of rare diseases 2022 — PMC9594925.
- A Novel Disulfidptosis-Related Diagnostic Gene Signature and Differential Expression Validation in Ischaemic Cardiomyopathy. Journal of cellular and molecular medicine 2025 — PMC11897062.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:19752196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NUBPL (Iron-sulfur cluster transfer protein NUBPL). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A10634-1). Validated IHC antibody: A10634-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Ciliated cells (tip of cilia)); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A10634-1); 2. Fixation: Image fixative and duration unreported (datasheet A10634-1); 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 A10634-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A10634-1); 8. Primary antibody: Rabbit anti-NUBPL, 2-5 μg/ml (datasheet A10634-1); 9. Primary incubation: Overnight at 4 °C (datasheet A10634-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences are unspecified (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). Evidence: UniProt Q8TB37, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the mitochondria., Datasheet A10634-1, PMC5480060, PMC11407088. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NUBPL/immunohistochemistry-ihc-guide.