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NDUFS8 / NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, mitochondrial · IHC design guide
Design Immunohistochemistry for NDUFS8
Plan NDUFS8 paraffin-section IHC around cytoplasmic staining in heart and skeletal muscle cells (HPA tissue IHC). Use those tissues to assess the expected pattern, while recognizing that the protein resides at the mitochondrial inner membrane (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NDUFS8 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 | Cardiomyocytes and myocytes show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08275-2) | |
| Positive control | Heart muscle+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 sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Highest expression in heart and skeletal muscle (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans 35–210 (UniProt) |
Section 1
Recommended NDUFS8 IHC & IF Protocols
The catalog antibody protocol is accompanied by published IHC methods for cardiomyocyte tissue (PMC13553469) and lung cancer tissue microarrays (PMC4981865).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet A08275-2) |
| Fixation | Image fixative and duration unreported (datasheet A08275-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A08275-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08275-2) |
| Primary antibody | Rabbit anti-NDUFS8, 2-5 μg/ml (datasheet A08275-2) |
| Primary incubation | Overnight at 4 °C (datasheet A08275-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08275-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NDUFS8-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues, including skeletal and heart muscle. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A08275-2); the published methods provide alkaline pH 9.0 and TRIS-EDTA alternatives (PMC13553469; PMC4981865).
Section 2
What Is the Expected NDUFS8 Staining Pattern?
NDUFS8 is a mitochondrial inner-membrane Complex I subunit with no transmembrane segment (UniProt O00217 topology). In paraffin-section IHC, expect cytoplasmic staining, especially in cardiomyocytes and skeletal-muscle myocytes (HPA: cytoplasmic profile; High in both cell types). HPA rates the tissue IHC pattern Enhanced, with medium consistency between staining and RNA expression (HPA: reliability).
| Strong cytoplasmic staining in cardiomyocytes or skeletal-muscle myocytes. | This matches the best-supported positive tissues and cell types (HPA: High in heart and skeletal muscle). A granular appearance can fit mitochondrial localisation, but chromogenic IHC need not resolve individual mitochondria (UniProt O00217 subcellular location). |
| Predominantly nuclear staining, with little cytoplasmic signal. | That compartment conflicts with mitochondrial inner-membrane localisation (UniProt O00217 topology; HPA: mitochondrial ICC-IF). Treat it as suspect until a known-positive tissue shows the expected cytoplasmic pattern. |
| Strong staining chiefly in adipocytes while nearby expected-positive myocytes are weak. | This reverses the reported pattern: adipocytes were not detected, whereas myocytes were High (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; a single discordant field does not establish either cause. |
| Uniform brown haze across cells and extracellular areas. | Diffuse background obscures the cell-restricted cytoplasmic pattern reported by HPA (HPA: tissue IHC profile). Examine the no-primary control and detection chemistry before assigning NDUFS8 positivity. |
| No staining in well-preserved heart or skeletal muscle. | This is unexpected for those high-staining cell types (HPA: High in cardiomyocytes and myocytes). First assess section quality and control performance, then revisit retrieval, antibody dilution and detection. |
💡Expected NDUFS8 appearanceCall a result positive when cardiomyocytes or skeletal-muscle myocytes show strong cytoplasmic staining (HPA: High in both); nuclear-only staining or diffuse extracellular colour is suspect given mitochondrial localisation (UniProt O00217; HPA: ICC-IF).
| Compartment and resolution | NDUFS8 resides at the mitochondrial inner membrane (UniProt O00217 topology), while tissue IHC is described as cytoplasmic (HPA: tissue profile); score the cytoplasmic cell pattern without requiring visibly discrete mitochondria. |
| Tissue comparison | Heart and skeletal muscle are High; listed glandular, respiratory epithelial and cerebellar cells are Medium (HPA: tissue IHC). Compare the named cell types, since whole-tissue averages can hide this variation. |
| Weak or absent comparator tissues | Lung alveolar cells and adipocytes were not detected, while liver hepatocytes were Low (HPA: tissue IHC). Use these as relative staining comparators, not proof that every cell in those tissues lacks protein. |
| Evidence strength | The tissue IHC reliability is Enhanced, yet staining and RNA have only medium consistency (HPA: reliability). HPA018524 and HPA067429 each have Enhanced IHC validation (HPA: antibodies); discordant specimens still need controls. |
| Processing and isoforms | UniProt lists a processed chain spanning residues 35–210 and no isoforms (UniProt O00217 processing; isoforms). The supplied record gives no antibody epitope, so it cannot predict which processing state an IHC reagent detects. |
| IF/ICC Q: what pattern belongs on its separate guide page? | A: Mitochondrial localisation is approved in ICC-IF (HPA: subcellular; HPA018524 ICC Approved). This supports compartment interpretation here; the supplied evidence gives no IF protocol option for this IHC guide. |
| Antigen retrieval and fixation | Retrieval is a general paraffin-IHC optimisation step. Neither HPA nor UniProt supplies an NDUFS8-specific fixation effect or retrieval setting; validate any change against positive tissue and controls without attributing it to target-specific masking. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart and skeletal muscle are both blank. | A shared staining-run failure is plausible because cardiomyocytes and myocytes are High in HPA tissue IHC (HPA: tissue IHC). | Check section integrity, the IHC-validated antibody conditions, retrieval, detection reagent and positive control; change one general workflow variable at a time. |
| Signal is confined to nuclei. | Nuclear-only staining conflicts with the reported mitochondrial location (UniProt O00217; HPA: ICC-IF). | Compare with a known-positive muscle section and a no-primary control, then reassess antibody specificity and image interpretation. |
| Adipocytes stain strongly but myocytes do not. | The pattern conflicts with HPA's Not detected adipocytes and High myocytes (HPA: tissue IHC); cross-reactivity or endogenous activity is possible. | Run a no-primary control and inspect cell identity and morphology before calling the adipocyte signal NDUFS8. |
| Broad brown background makes cell boundaries hard to score. | Nonspecific detection or endogenous activity may obscure the expected cytoplasmic pattern (HPA: tissue IHC profile). | Review the no-primary control, blocking, washes and detection exposure; score only a reproducible cell-associated pattern. |
| A lung section appears negative. | Alveolar cells were Not detected in HPA tissue IHC, and UniProt reports the lowest tissue expression in lung (HPA: lung; UniProt O00217 tissue specificity). | Do not diagnose assay failure from lung alone; include heart or skeletal muscle as the positive tissue control (HPA: High). |
| Retrieval changes the apparent staining strength. | Retrieval conditions can affect paraffin-IHC performance in general; no NDUFS8-specific fixation or retrieval sensitivity is supplied. | Compare conditions on matched sections with positive tissue and no-primary controls, and report the observed change without claiming a target-specific fixation mechanism. |
Experiment evidence
IHC & IF images showing NDUFS8 staining
A08275-2 · IHCIHC analysis of NDUFS8 using anti-NDUFS8 antibody (A08275-2). NDUFS8 was detected in a paraffin-embedded section of human liver cancer tissu
A08275-2 · IHCIHC analysis of NDUFS8 using anti-NDUFS8 antibody (A08275-2). NDUFS8 was detected in a paraffin-embedded section of mouse heart tissue. Heat
A08275-2 · IFIF analysis of NDUFS8 using anti-NDUFS8 antibody (A08275-2). NDUFS8 was detected in a paraffin-embedded section of human liver cancer tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000110717-NDUFS8/tissue/cerebral+cortex#imid_4830760
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000110717-NDUFS8/tissue/colon#imid_4830670
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000110717-NDUFS8/tissue/liver#imid_4830671
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000110717-NDUFS8/tissue/kidney#imid_4830688
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018524; green = NDUFS8, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018524; green = NDUFS8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NDUFS8 IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain strongly (HPA: High in cardiomyocytes); use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes). On the positive slide, cardiomyocyte nuclei and extracellular spaces should lack specific staining (UniProt O00217: mitochondrial inner membrane localization).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NDUFS8 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls (selected-SKU IHC caption: rabbit primary at 2 μg/ml), plus a NDUFS8 knockout sample as a biological specificity control (standard IHC practice). Block endogenous peroxidase in heart sections before HRP/DAB detection (selected-SKU IHC caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A08275-2 tissue-IHC caption does not state a fixative (selected-SKU IHC caption). That caption reports heat-mediated retrieval in EDTA at pH 8.0 before staining, but does not establish whether retrieval is required under other conditions (selected-SKU IHC caption). ICC-IF images support mitochondrial localization, but the supplied evidence does not show that frozen sections or IF are easier; endogenous peroxidase can complicate interpretation of HRP/DAB staining in heart muscle (HPA: mitochondria, approved; standard IHC practice).
HPA tissue IHC evidence for NDUFS8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NDUFS8 IHC Tips
Troubleshoot NDUFS8 staining in paraffin sections using the catalog antibody’s IHC result, mitochondrial localisation, and tissue expression as reference points.
What retrieval should I try first if NDUFS8 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A08275-2). The catalog antibody produced staining in a paraffin-embedded human liver cancer section after that retrieval (datasheet A08275-2). Include heart or skeletal muscle as a positive control, since cardiomyocytes and myocytes show high staining (HPA: High in cardiomyocytes and myocytes). If staining remains weak, adjust heating time or cooling conditions on matched sections while holding antibody concentration and detection constant; these are optimisation trials, not established NDUFS8 requirements. Compare mitochondrial-pattern signal with tissue damage and background before accepting a stronger retrieval condition (UniProt O00217: mitochondrial inner membrane).
How should I assess whether fixation is affecting NDUFS8 IHC?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (datasheet A08275-2: fixative not stated). Record fixation method, duration, tissue thickness, and processing history for each specimen, then compare similarly processed sections under the same retrieval and detection conditions (standard IHC practice). Keep the documented starting retrieval at EDTA pH 8.0 during that comparison (datasheet A08275-2). Use a heart or skeletal-muscle control to check whether a run can reveal NDUFS8 where staining is high (HPA: High in cardiomyocytes and myocytes). A weak specimen beside a strong control raises a processing or biological question; it does not establish fixation as the cause.
Where should convincing NDUFS8 staining appear in a tissue section?
Look for cytoplasmic staining compatible with mitochondria, while recognising that chromogenic IHC may not resolve individual organelles (HPA: cytoplasmic tissue expression; HPA: mitochondria approved). NDUFS8 is assigned to the mitochondrial inner membrane and has no predicted transmembrane segment (UniProt O00217: subcellular location and topology). Heart cardiomyocytes and skeletal-muscle myocytes offer strong tissue references; hepatocytes have low staining in the listed tissue survey (HPA: High in cardiomyocytes and myocytes; Low in hepatocytes). Compare the distribution within intact cells against those references and against the section’s counterstain (standard IHC practice). Predominantly nuclear or extracellular deposits do not fit the reported location and warrant review of controls and morphology (HPA: mitochondria approved).
Could isoforms or epitope accessibility explain variable NDUFS8 staining?
The supplied record lists 0 isoforms and a processed chain spanning residues 35–210 (UniProt O00217: isoforms and processing). It also lists no glycosylation sites or modified residues, so those annotations provide no specific explanation for a staining difference (UniProt O00217: glycosylation and modified residues). The antibody’s recognised sequence is not supplied; do not assign its epitope to either the processed chain or the precursor. Compare matched sections with the documented EDTA pH 8.0 retrieval and 2 μg/ml primary concentration before changing conditions (datasheet A08275-2). If accessibility remains in question, interpret a retrieval response as assay behaviour, not proof of an isoform or modification.
How can I check NDUFS8 localisation by IF alongside tissue IHC?
Use IF as a complementary localisation check, keeping the chromogenic paraffin-section result as the IHC reference (datasheet A08275-2). Multiplex NDUFS8 with a validated marker for cardiomyocytes or skeletal-muscle myocytes, which show high tissue staining (HPA: High in cardiomyocytes and myocytes). Choose a fluorophore channel after inspecting unstained tissue for autofluorescence, and include single-colour controls when separating multiplex signals (standard IF practice). NDUFS8 is assigned to the mitochondrial inner membrane, but the antibody epitope and its membrane-facing side are unspecified (UniProt O00217: location and topology; datasheet A08275-2: epitope not stated). Compare permeabilisation conditions empirically; a missing IF signal alone cannot identify which side of the membrane contains the epitope.
How do I distinguish NDUFS8 signal from chromogenic background?
Inspect a section without primary antibody alongside the stained section to assess secondary-reagent and chromogen background (standard IHC practice). The documented workflow used 10% goat serum blocking, 2 μg/ml rabbit primary overnight at 4°C, and an anti-rabbit peroxidase secondary for 30 minutes at 37°C (datasheet A08275-2). If diffuse colour obscures cell detail, review blocking, washes, primary exposure, and DAB development one variable at a time (standard IHC practice). Include a peroxidase-blocking step and its control as general chromogenic IHC practice; neither establishes NDUFS8 specificity. Judge retained staining against mitochondrial-compatible cytoplasmic distribution and intact morphology (HPA: mitochondria approved; HPA: cytoplasmic tissue expression).
What is a defensible way to quantify NDUFS8 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and region before comparing slides; cardiomyocytes, skeletal-muscle myocytes, and hepatocytes have different reported staining levels (HPA: High in cardiomyocytes and myocytes; Low in hepatocytes). For cell-level chromogenic staining, report an H-score from intensity categories and the percentage of positive cells, using identical thresholds across slides (standard IHC scoring practice). If counting stained cells, report density per mm² of evaluable tissue and exclude folds, necrosis, and tissue-free areas (standard IHC scoring practice). Normalise to the number of eligible cells or evaluable tissue area, and compare matched compartments and processing conditions. Keep the positive-control result visible when interpreting between-run differences (HPA: High in cardiomyocytes and myocytes).
When should an apparent NDUFS8-positive area be treated as artefact?
A credible result follows intact-cell cytoplasmic distribution compatible with mitochondria, with the expected strong staining in cardiomyocytes or skeletal-muscle myocytes (HPA: cytoplasmic tissue expression; HPA: High in cardiomyocytes and myocytes). Treat predominantly nuclear or extracellular colour as discordant with the assigned mitochondrial inner-membrane location (UniProt O00217: subcellular location). Check whether signal concentrates at section edges, folds, or necrotic areas before scoring it as cellular staining (standard IHC practice). Compare a no-primary section and peroxidase-block control to assess detection background and endogenous enzyme colour (standard chromogenic IHC practice). A negative hepatocyte area alone is inconclusive because hepatocyte staining is reported as low (HPA: Low in hepatocytes).
Boster reagents
Best NDUFS8 / NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, mitochondrial IHC Antibodies
The IHC-validated antibody A08275-2 has IHC images from human liver cancer and mouse heart paraffin sections, plus IF data from a human liver cancer paraffin section (catalog image captions).
Real IHC data
Anti-NDUFS8 Antibody ®
Cat # A08275-2
A08275-2 has IHC images from human liver cancer and mouse heart paraffin sections (catalog IHC captions). Its IF image uses a human liver cancer paraffin section; listed reactivity covers human, mouse, and rat (catalog IF caption; catalog reactivity).
Which to pick: For tissue IHC, choose A08275-2: its rabbit antibody was used at 2 μg/ml in human and mouse paraffin sections (catalog IHC captions); the fixative is unreported (catalog IHC captions). For IF, the same SKU has a human paraffin-section example at 5 μg/ml (catalog IF caption); ICC validation and clonality are unreported (catalog applications and clone field). For cross-species work, A08275-2 lists human, mouse, and rat reactivity, with IHC images for human and mouse only (catalog reactivity; catalog IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00217 (NDUS8_HUMAN, NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, mitochondrial).
- Human Protein Atlas. NDUFS8 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NDUFS8 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. NDUFS8 antibody validation summary (2 antibodies).
- Combined machine learning identification and experimental validation of NDUFS8 and SUCLG1 as potential biomarkers for mechanical asphyxia. Frontiers in medicine 2026 — PMC13553469.
- NADH:ubiquinone oxidoreductase core subunit S8 expression and functional significance in non-small cell lung cancer. Cell death & disease 2025 — PMC12012183.
- The opposite prognostic effect of NDUFS1 and NDUFS8 in lung cancer reflects the oncojanus role of mitochondrial complex I. Scientific reports 2016 — PMC4981865.
- Human umbilical mesenchymal stem cells ameliorate atrophic gastritis in aging mice by participating in mitochondrial autophagy through Ndufs8 signaling. Stem cell research & therapy 2024 — PMC11662822.
- PubMed PMID:9116042 — UniProt-cited evidence.
- PubMed PMID:9666055 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NDUFS8 (NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, mitochondrial). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08275-2). Validated IHC antibody: A08275-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); 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 A08275-2); 2. Fixation: Image fixative and duration unreported (datasheet A08275-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A08275-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08275-2); 8. Primary antibody: Rabbit anti-NDUFS8, 2-5 μg/ml (datasheet A08275-2); 9. Primary incubation: Overnight at 4 °C (datasheet A08275-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans 35–210 (UniProt); Regulation: Highest expression in heart and skeletal muscle (UniProt). Evidence: UniProt O00217, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A08275-2, PMC13553469, PMC4981865. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NDUFS8/immunohistochemistry-ihc-guide.