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NDUFC1 / NADH dehydrogenase [ubiquinone] 1 subunit C1, mitochondrial · IHC design guide
Design Immunohistochemistry for NDUFC1
Plan paraffin IHC for NDUFC1 with the catalog antibody at 1:100–1:300 (datasheet A15096-1). Use the cytoplasmic tissue profile and high staining in kidney tubules to assess the result (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NDUFC1 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 staining in tissue (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A15096-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Bone marrow+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 | IHC staining has medium agreement with RNA data (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | 0 isoforms; mature chain 28–76; epitope unknown (UniProt) |
Section 1
Recommended NDUFC1 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A15096-1) is accompanied by two published NDUFC1 IHC protocols (PMC8710466; PMC9479186).
| Sample | Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A15096-1) |
| Fixation | Image fixative and duration unreported (datasheet A15096-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A15096-1); 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-NDUFC1, 1:100-1:300 (datasheet A15096-1) |
| 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 | NDUFC1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with pH 9.0 Tris-EDTA heat retrieval (datasheet A15096-1); compare the published citric acid conditions if staining needs optimization (PMC8710466; PMC9479186).
Section 2
What Is the Expected NDUFC1 Staining Pattern?
NDUFC1 is an inner mitochondrial membrane Complex I subunit with a transmembrane segment at residues 41–59 (UniProt O43677 topology). In paraffin IHC, expect cytoplasmic staining in cell populations reported positive by HPA, including kidney tubular and colon glandular cells (HPA tissue IHC: High). HPA calls the tissue pattern ubiquitous cytoplasmic, with Enhanced reliability, medium agreement with RNA expression, and external verification pending (HPA tissue IHC).
| Granular cytoplasmic staining in kidney tubular cells or colon glandular cells, with little distracting background. | This fits the mitochondrial location and the cell populations scored High by HPA (UniProt O43677 subcellular location; HPA tissue IHC: High in kidney tubular cells and colon glandular cells). Chromogenic IHC shows a cellular pattern; it cannot resolve staining specifically to the inner mitochondrial membrane (general IHC practice). |
| Predominantly nuclear or cell-surface staining in otherwise positive cells. | That compartment conflicts with the reported mitochondrial location (UniProt O43677 subcellular location; HPA subcellular ICC-IF: mitochondria). Treat it as a possible assay artefact and inspect controls before scoring it as NDUFC1 (general IHC practice). |
| Strong staining in a cell population HPA lists as not detected, such as bone marrow hematopoietic cells. | This is discordant with the reported cell-specific pattern (HPA tissue IHC: Not detected in bone marrow hematopoietic cells). Consider antibody cross-reactivity or endogenous chromogenic activity, and assess the staining against appropriate controls (general IHC practice). |
| Diffuse colour covers cells, extracellular spaces, or the whole section without a discernible cytoplasmic pattern. | The result cannot be interpreted as the reported cytoplasmic pattern (HPA tissue IHC: ubiquitous cytoplasmic expression). Uneven reagent deposition, nonspecific binding, or detection background are possible causes; compare with a detection control (general IHC practice). |
| No staining in kidney tubular cells or colon glandular cells on a run expected to show NDUFC1. | Those populations are reported High and can serve as positive comparators (HPA tissue IHC: High in kidney tubular cells and colon glandular cells). First check whether the run and tissue are interpretable; absence alone does not establish biological loss of NDUFC1 (general IHC practice). |
💡Expected NDUFC1 appearanceCall a result positive when expected cell populations show convincing cytoplasmic staining, especially HPA High kidney tubules or colon glands; isolated nuclear or extracellular colour is suspect (HPA tissue IHC; UniProt O43677 subcellular location; general IHC practice).
| Compartment and optical resolution | NDUFC1 occupies the mitochondrial inner membrane, with one annotated transmembrane segment at 41–59 (UniProt O43677 topology). Interpret chromogenic signal at the cytoplasmic level; the slide cannot establish membrane topology (general IHC practice). |
| Processing and epitope limits | UniProt annotates a mature chain at residues 28–76 and no glycosylation sites or modified residues (UniProt O43677 processing/PTMs). No antibody epitope is supplied, so these annotations cannot predict antigen retrieval needs or staining sensitivity. |
| Cell-specific tissue comparison | HPA scores kidney tubular and colon glandular cells High, but bone marrow hematopoietic cells and oral mucosal squamous cells Not detected (HPA tissue IHC). Compare the named cell populations, since a tissue-wide positive or negative label would overstate the evidence. |
| Strength of IHC evidence | Both listed antibodies, HPA044556 and HPA062427, have Enhanced IHC status (HPA antibodies). The tissue profile reports medium agreement with RNA and pending external verification (HPA tissue IHC); use the pattern as an expectation, not proof of specificity in every section. |
| IF/ICC Q: What localization should appear? | A: Mitochondria are the supported main location; HPA lists ICC-IF images in A-431, U-251MG, and U2OS and marks HPA044556 ICC Approved (HPA subcellular ICC-IF; HPA antibodies). This localization cross-check does not specify an IF protocol for this IHC guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparator has no convincing cytoplasmic signal. | The run may have weak detection, an unsuitable antibody concentration, or a missed processing step (general IHC practice); the expected populations are High (HPA tissue IHC: kidney tubular and colon glandular cells). | Check the positive control, primary-antibody application, detection reagents, and counterstain. Optimize retrieval and antibody concentration as general IHC variables; target-specific fixation or retrieval sensitivity is unreported in the supplied sources. |
| Nuclei dominate the staining. | The distribution conflicts with the mitochondrial assignment (UniProt O43677 subcellular location; HPA subcellular ICC-IF). Excess background or nonspecific binding may obscure the intended pattern (general IHC practice). | Compare with the primary-omission control, review antibody concentration and blocking, and score only a reproducible cytoplasmic pattern in an expected cell population (general IHC practice; HPA tissue IHC). |
| Strong signal appears in HPA Not detected cells. | Unexpected cell-specific staining may reflect cross-reactivity or endogenous detection activity (general IHC practice; HPA tissue IHC: Not detected in bone marrow hematopoietic cells). | Confirm which cells carry the colour, then inspect primary-omission and detection controls. If signal persists without primary antibody, address endogenous detection activity before interpreting the section (general IHC practice). |
| Diffuse colour obscures cell boundaries. | Nonspecific binding, inadequate washing, or excess detection product can obscure a cytoplasmic pattern (general IHC practice; HPA tissue IHC: ubiquitous cytoplasmic expression). | Review blocking and wash steps, antibody concentration, and chromogen development using the same run's controls. Reassess whether staining follows identifiable cells before assigning intensity (general IHC practice). |
| Only low-level or variable staining appears across candidate tissues. | HPA reports Low staining in several named populations, including thyroid glandular and bronchial respiratory epithelial cells; weak signal there is less informative than a High comparator (HPA tissue IHC). | Read the correct cell population alongside kidney tubules or colon glands, and avoid treating every faint deposit as positive. Record intensity and background together when scoring (HPA tissue IHC; general IHC practice). |
| A negative cell population is used to declare the entire section negative. | HPA levels refer to specified cell populations: ovarian stroma is Not detected, while the overall tissue profile is described as ubiquitous cytoplasmic expression (HPA tissue IHC). | Identify the cells being scored and report their staining separately. Use a named High population as the positive comparator before drawing a conclusion about assay performance (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing NDUFC1 staining
A15096-1 · IHCImmunohistochemical analysis of paraffin-embedded human Colon cancer. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109390-NDUFC1/tissue/cerebral+cortex#imid_20843559
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109390-NDUFC1/tissue/colon#imid_20843485
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109390-NDUFC1/tissue/liver#imid_20843486
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109390-NDUFC1/tissue/kidney#imid_20843495
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044556; green = NDUFC1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044556; green = NDUFC1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NDUFC1 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use parathyroid glandular cells as a negative tissue (HPA: Not detected); on the colon slide, assess non-glandular stromal cells for background staining as an internal comparator, without assuming they lack NDUFC1 (standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NDUFC1 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus NDUFC1 knockout material or a cognate-peptide block to test specificity (standard IHC practice). Quench endogenous peroxidase in colon sections when using chromogenic peroxidase detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A15096-1 paraffin-section caption does not state a fixative (selected IHC caption: fixative not stated). The caption uses Tris-EDTA at pH 9.0 for retrieval, but does not establish that retrieval is required; it reports a 1:200 primary dilution overnight at 4°C in colon cancer tissue (selected IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; interpret any cytoplasmic staining against the expected mitochondrial location and monitor colon tissue for endogenous peroxidase signal (UniProt O43677: mitochondrial inner membrane; standard IHC practice).
HPA tissue IHC evidence for NDUFC1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NDUFC1 IHC Tips
Troubleshoot NDUFC1 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
How should I retrieve NDUFC1 in paraffin sections with weak staining?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 (datasheet A15096-1). The catalog antibody's paraffin-section image used this buffer with a 1:200 primary dilution overnight at 4°C, but the caption gives no heating duration (datasheet A15096-1). Run a retrieval time series on adjacent sections, keeping the antibody dilution and detection conditions fixed, and check both signal and tissue morphology (standard IHC practice). If staining remains weak, test an alternative retrieval buffer only as a fallback, with positive and negative controls processed alongside it; avoid interpreting stronger background as improved NDUFC1 detection (standard IHC practice).
Could fixation explain weak or uneven NDUFC1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report a fixative (datasheet A15096-1). Record the actual fixative, fixation duration and processing history for each specimen before comparing staining intensity across blocks (standard IHC practice). On serial sections from the same block, keep Tris-EDTA retrieval at pH 9.0 and the detection settings constant while checking whether weak regions coincide with damaged morphology or processing gradients (datasheet A15096-1; standard IHC practice). Treat any fixation effect as a specimen-level hypothesis until controlled comparisons support it; neither the tissue staining profile nor the protein's membrane topology establishes fixation sensitivity (HPA: tissue profile; UniProt O43677: topology).
What staining pattern should count as NDUFC1 in tissue?
Assess signal within the cytoplasm, where a mitochondrial distribution is expected for this inner-membrane Complex I subunit (UniProt O43677: location and function; HPA: supported mitochondrial location). The tissue atlas describes ubiquitous cytoplasmic expression, so a diffuse chromogenic appearance alone does not establish which organelle generated the signal (HPA: tissue profile). Compare glandular cells in colon with adjacent compartments, and examine whether staining is cellular rather than confined to section folds, edges or luminal debris (HPA: High in colon glandular cells; standard IHC practice). A predominantly nuclear or extracellular pattern conflicts with the supported mitochondrial assignment and warrants review of controls, morphology and detection background (HPA: supported mitochondrial location; standard IHC practice).
How does NDUFC1 topology affect epitope accessibility?
NDUFC1 has a 76-amino-acid precursor, an annotated chain spanning residues 28–76, and a transmembrane segment at residues 41–59 (UniProt O43677: processing and topology). No isoforms, glycosylation sites or modified residues are annotated in the supplied record, so those features cannot explain a staining difference without further evidence (UniProt O43677: isoforms and modifications). The catalog caption does not identify the antibody epitope; do not assign it to either side of the inner membrane from the protein sequence alone (datasheet A15096-1; UniProt O43677: topology). If staining changes with retrieval, compare controlled retrieval conditions and seek epitope information before attributing the change to processing or membrane accessibility (standard IHC practice).
How can IF help assess NDUFC1 localisation?
Use IF as a separate localisation check: the atlas reports supported mitochondrial localisation and lists ICC/IF images from A-431, U-251MG and U2OS cells (HPA: subcellular profile). For tissue multiplexing, pair NDUFC1 with a marker identifying the expected cell type in the section, and assess mitochondrial pattern within those cells rather than relying on total fluorescence (standard IF practice; HPA: supported mitochondrial location). Select fluorophores and matched unstained controls around the specimen's autofluorescence spectrum, especially before treating faint signal as positive (standard IF practice). Because the antibody epitope and its membrane-facing side are unspecified, optimise permeabilisation empirically and do not assume that a condition exposes a particular side of the inner membrane (datasheet A15096-1; UniProt O43677: topology; standard IF practice).
How do I distinguish NDUFC1 signal from chromogenic background?
Compare the IHC-validated antibody at the documented 1:200 dilution with a no-primary control on adjacent paraffin sections (datasheet A15096-1; standard IHC practice). Apply a peroxidase block before DAB detection and check whether pigment or endogenous enzyme activity persists in the no-primary section; these are general chromogenic IHC controls, not NDUFC1-specific validation (standard IHC practice). If background is broad, optimise blocking, washing, primary exposure and detection strength one variable at a time while preserving the documented Tris-EDTA retrieval at pH 9.0 (datasheet A15096-1; standard IHC practice). Compare staining within intact cell boundaries, since folds, edges and damaged regions can accumulate misleading chromogen (standard IHC practice).
How should I quantify NDUFC1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis region before scoring, using intact cells and a consistent chromogen threshold across sections (standard IHC practice). For glandular compartments, report the percentage of positive cells and an H-score from intensity categories; keep cell counts and region area available for review (standard IHC practice; HPA: High in colon glandular cells). If counting positive cells per mm², normalise to the analysed viable tissue area and report cellular composition, since a change in glandular-cell abundance can alter the section-wide result (standard IHC practice). Use identically processed reference sections and compare like compartments, because the atlas reports both high and undetected staining among specified cell populations (HPA: tissue profile; standard IHC practice).
When is an apparent NDUFC1-positive area likely to be artefact?
Interpret signal as plausible NDUFC1 only when it follows intact cellular cytoplasm and is consistent with mitochondrial localisation (HPA: ubiquitous cytoplasmic expression and supported mitochondrial location; UniProt O43677: inner-membrane location). Check cell identity: high staining is reported in colon glandular cells, whereas bone-marrow hematopoietic cells are listed as not detected in the atlas (HPA: tissue profile). Nuclear or extracellular deposits, edge-concentrated staining and signal over necrotic areas need control and morphology review before scoring (HPA: supported mitochondrial location; standard IHC practice). Persistent DAB colour in a no-primary control can indicate endogenous enzyme activity or detection background, so exclude that pattern from target-specific interpretation (standard IHC practice).
Boster reagents
Best NDUFC1 / NADH dehydrogenase [ubiquinone] 1 subunit C1, mitochondrial IHC Antibodies
The catalog antibody has real IHC data from paraffin-embedded human colon cancer (catalog IHC image caption); IF is listed (catalog applications), with human and mouse reactivity (catalog reactivity).
Real IHC data
Anti-NDUFC1 Antibody
Cat # A15096-1
A15096-1 is shown by IHC on paraffin-embedded human colon cancer using 1:200 primary dilution and Tris-EDTA pH 9.0 retrieval (catalog IHC image caption). A15096-1 also lists IF and human/mouse reactivity, but provides no IF image (catalog applications/reactivity; catalog IF image alts).
Which to pick: For tissue IHC, choose A15096-1 because its own image shows staining in paraffin-embedded human colon cancer; the fixative is unreported (catalog IHC image caption). For IF/ICC, A15096-1 lists IF at 1:50, but ICC-specific validation and an IF image are unreported (catalog applications/dilution; catalog IF image alts). For work across human and mouse, A15096-1 lists both species, although the supplied IHC image shows human tissue only and clonality is unreported (catalog reactivity; catalog IHC image caption; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43677 (NDUC1_HUMAN, NADH dehydrogenase [ubiquinone] 1 subunit C1, mitochondrial).
- Human Protein Atlas. NDUFC1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NDUFC1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. NDUFC1 antibody validation summary (2 antibodies).
- NDUFC1 Is Upregulated in Gastric Cancer and Regulates Cell Proliferation, Apoptosis, Cycle and Migration. Frontiers in oncology 2021 — PMC8710466.
- Knockdown of NDUFC1 inhibits cell proliferation, migration, and invasion of hepatocellular carcinoma. Frontiers in oncology 2022 — PMC9479186.
- PubMed PMID:9425316 — UniProt-cited evidence.
- PubMed PMID:9653160 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NDUFC1 (NADH dehydrogenase [ubiquinone] 1 subunit C1, mitochondrial). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A15096-1). Validated IHC antibody: A15096-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Bone marrow. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Colon cancer tissue; fixative not specified (datasheet A15096-1); 2. Fixation: Image fixative and duration unreported (datasheet A15096-1); 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: Tris-EDTA pH 9.0 (datasheet A15096-1); 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-NDUFC1, 1:100-1:300 (datasheet A15096-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: IHC staining has medium agreement with RNA data (HPA tissue IHC); Isoform / epitope: 0 isoforms; mature chain 28–76 ; epitope unknown (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt O43677, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A15096-1, PMC8710466, PMC9479186. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NDUFC1/immunohistochemistry-ihc-guide.