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NDUFV2 / NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial · IHC design guide
Design Immunohistochemistry for NDUFV2
Plan paraffin-section IHC for NDUFV2 using its granular cytoplasmic tissue pattern (HPA tissue IHC) and inner-membrane location (UniProt). Use the catalog antibody’s IHC evidence to guide detection and interpret staining alongside controls.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NDUFV2 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 | Granular cytoplasmic staining (HPA tissue IHC); inner membrane at the molecular level (UniProt) | |
| Staining pattern | Granular cytoplasm in cardiomyocytes and glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05801-2) | |
| Positive control | Bronchus+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 paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; mature chain begins at residue 33 (UniProt) |
Section 1
Recommended NDUFV2 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet A05801-2). Published paraffin section protocols cover rat brain (PMC8374556) and human lung adenocarcinoma tissue (PMC12847814).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A05801-2) |
| Fixation | Image fixative and duration unreported (datasheet A05801-2); 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 A05801-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05801-2) |
| Primary antibody | Rabbit anti-NDUFV2, 2-5 μg/ml (datasheet A05801-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05801-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05801-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NDUFV2-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A05801-2); the rat brain protocol also used heated EDTA retrieval (PMC8374556).
Section 2
What Is the Expected NDUFV2 Staining Pattern?
NDUFV2 is a Complex I subunit of the mitochondrial inner membrane with no annotated transmembrane segment (UniProt P19404). In paraffin-section IHC, expect predominantly granular cytoplasmic staining, especially in respiratory epithelium, intestinal glandular cells, neurons, and cardiomyocytes (HPA tissue IHC). HPA rates the tissue staining profile Enhanced, while describing only medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Granular cytoplasmic staining in bronchial respiratory epithelium, colonic glands, or cardiomyocytes. | This matches the reported high staining in those cell types and the general granular cytoplasmic profile (HPA tissue IHC). The granules are consistent with mitochondrial localisation (UniProt P19404); IHC morphology alone does not identify individual organelles. |
| Predominantly nuclear staining, with little granular cytoplasmic signal, in an expected-positive tissue. | Treat a nuclear-only IHC pattern as discordant with the reported tissue profile (HPA tissue IHC). Nucleoplasm is an additional approved ICC-IF location (HPA subcellular), so investigate the assay before calling every nuclear signal an artefact. |
| Strong staining of adipocytes, ovarian stromal cells, or splenic red-pulp cells. | These cell populations were reported as not detected (HPA tissue IHC). Check cell identification and controls; unexpected staining may reflect cross-reactivity or endogenous detection activity (standard IHC practice), but the slide alone cannot distinguish them. |
| Diffuse colour across cells and extracellular areas, obscuring cytoplasmic granules. | This does not resemble the reported granular cytoplasmic pattern (HPA tissue IHC). Review background in a no-primary control and the detection steps before interpreting weak cell staining (standard IHC practice). |
| No visible signal in bronchial respiratory epithelium or heart cardiomyocytes. | Both are reported as high-staining cell types (HPA tissue IHC). Confirm that the expected cells are present and compare a known-positive section processed in the same run; a blank result may indicate an assay failure (standard IHC practice). |
💡Expected NDUFV2 appearanceCall positive a predominantly granular cytoplasmic signal with high staining in an HPA high cell type, such as cardiomyocytes (HPA tissue IHC); isolated nuclear colour or broad cell-independent haze warrants control review (standard IHC practice).
| Compartment and topology | NDUFV2 belongs to mitochondrial inner-membrane Complex I and has no annotated transmembrane segment (UniProt P19404). The tissue IHC profile is granular cytoplasmic (HPA tissue IHC); topology alone does not predict a retrieval condition. |
| Choice of positive tissue | Respiratory epithelial cells in bronchus, glandular cells in colon and duodenum, and heart cardiomyocytes are reported High (HPA tissue IHC). Select a section containing a clearly identifiable listed cell population. |
| Choice of comparison cells | Adipocytes, bone-marrow hematopoietic cells, ovarian stromal cells, fibroblasts in soft tissue, and splenic red-pulp cells are reported Not detected (HPA tissue IHC). Use these as pattern comparisons, not proof of absolute biological absence. |
| Strength of evidence | The tissue profile is rated Enhanced but has medium staining-to-RNA consistency (HPA tissue IHC). HPA003404 is IHC Supported and HPA077896 is IHC Enhanced (HPA antibodies); keep each antibody's validation status distinct from the tissue-profile rating. |
| Processing and epitope limits | UniProt lists a mature chain spanning residues 33–249 (UniProt P19404). No antibody epitope or target-specific fixation effect is supplied, so neither antigen-retrieval response nor loss of a particular epitope can be predicted from that processing annotation. |
| IF/ICC Q&A | Where should IF/ICC signal appear? Mainly in mitochondria, with additional approved nucleoplasmic and cytosolic locations (HPA subcellular). Those ICC-IF locations inform interpretation; they do not provide an IF protocol or override the tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells remain blank. | The result conflicts with High staining reported for bronchial respiratory epithelium and cardiomyocytes (HPA tissue IHC); the failed step cannot be identified from the blank slide alone. | Check cell identity and the same-run positive control, then review antibody application, retrieval, and detection one step at a time (standard IHC practice). |
| Colour is diffuse across the section. | Broad haze obscures the expected granular cytoplasmic profile (HPA tissue IHC); excess background can arise during blocking, antibody incubation, washing, or detection (standard IHC practice). | Compare a no-primary control, review washing and blocking, and adjust the working antibody concentration if controls implicate nonspecific binding (standard IHC practice). |
| Only nuclei appear positive. | Nuclear-only IHC is discordant with the reported general cytoplasmic profile (HPA tissue IHC), although nucleoplasm is an additional ICC-IF location (HPA subcellular). | Recheck the chromogenic controls and compare another expected-positive tissue cell type before assigning the nuclear colour to NDUFV2 (standard IHC practice; HPA tissue IHC). |
| Cells reported Not detected stain strongly. | Staining of adipocytes or ovarian stromal cells conflicts with the HPA cell-level observations (HPA tissue IHC); cross-reactivity and endogenous detection activity are possible assay causes (standard IHC practice). | Confirm the cell type, inspect a no-primary control, and review endogenous enzyme blocking where relevant to the chromogen (standard IHC practice). |
| Expected-positive cells show patchy or weak colour. | HPA reports High staining for specified cell populations, but its tissue profile has medium consistency with RNA data (HPA tissue IHC); variation alone does not identify a technical cause. | Compare matched positive sections from the same run and review section quality, retrieval, and detection consistency (standard IHC practice). Avoid inferring NDUFV2-specific fixation sensitivity. |
| A fluorescent image appears more nuclear than the chromogenic section. | HPA reports additional approved nucleoplasmic and cytosolic ICC-IF locations alongside the main mitochondrial location (HPA subcellular); tissue IHC is described as granular cytoplasmic (HPA tissue IHC). | Interpret each application against its own HPA pattern and controls; assess whether the apparent signal tracks cells or background before calling the images inconsistent (standard IHC practice). |
Experiment evidence
IHC & IF images showing NDUFV2 staining
A05801-2 · IHCIHC analysis of NDUFV2 using anti-NDUFV2 antibody (A05801-2). NDUFV2 was detected in a paraffin-embedded section of human colorectal adenoca
A05801-2 · IHCIHC analysis of NDUFV2 using anti-NDUFV2 antibody (A05801-2). NDUFV2 was detected in a paraffin-embedded section of human esophageal squamou
A05801-2 · IFIF analysis of NDUFV2 using anti-NDUFV2 antibody (A05801-2). NDUFV2 was detected in a paraffin-embedded section of human rectal cancer tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000178127-NDUFV2/tissue/cerebral+cortex#imid_1355852
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000178127-NDUFV2/tissue/colon#imid_1355778
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000178127-NDUFV2/tissue/liver#imid_1355779
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000178127-NDUFV2/tissue/kidney#imid_1355780
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA077896; green = NDUFV2, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA077896; green = NDUFV2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NDUFV2 IHC & IF
🧪Run bronchus first: respiratory epithelial cells should stain (HPA: High in respiratory epithelial cells). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the bronchus slide, cells without specific staining provide an internal background reference, but their target-negative status requires validation.
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NDUFV2 in Hep-G2, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality, and a biological negative such as NDUFV2 knockout tissue or a validated peptide-block control (caption: rabbit anti-NDUFV2 antibody). For chromogenic detection, quench endogenous peroxidase and check bronchus sections for background staining; for IF, inspect an unstained section for autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A05801-2 tissue-IHC caption does not state its fixative (caption: fixative not stated). The reported paraffin-section workflow used heat-mediated EDTA retrieval at pH 8.0, followed by primary antibody at 2 μg/ml overnight at 4°C (caption: retrieval and staining conditions). The supplied evidence does not establish that frozen sections or IF are easier; bronchus autofluorescence could complicate IF assessment (standard IF practice).
HPA tissue IHC evidence for NDUFV2
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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | 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 → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NDUFV2 IHC Tips
Use the paraffin-section IHC example and compartment-specific controls to troubleshoot NDUFV2 staining; interpret IF findings separately.
Which retrieval conditions should I try first for weak NDUFV2 staining?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A05801-2). The selected image used that retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A05801-2). If staining remains weak, compare a second retrieval condition on adjacent sections while keeping antibody concentration, detection and development constant (standard IHC practice). Include a known positive tissue, such as colon glandular cells, to distinguish retrieval failure from a genuinely faint specimen (HPA: High in colon glandular cells). Excess retrieval can damage morphology, so assess tissue integrity alongside staining intensity (standard IHC practice).
Could fixation explain loss of NDUFV2 staining in my sections?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A05801-2). Record each specimen’s fixative, fixation duration, processing history and section age before comparing signal across batches (standard IHC practice). If staining differs, process a known positive control alongside the specimens and vary one preparation factor at a time (standard IHC practice; HPA: High in colon glandular cells). Compare granular cytoplasmic staining and preserved morphology rather than judging intensity alone (HPA: granular cytoplasmic profile; standard IHC practice). Neither the tissue-IHC pattern nor the protein’s topology establishes how NDUFV2 responds to fixation (HPA: tissue IHC; UniProt P19404 topology).
What staining pattern is plausible for NDUFV2 in chromogenic sections?
Expect predominantly granular cytoplasmic staining at light-microscope resolution, consistent with mitochondrial localisation (HPA: granular cytoplasmic profile; UniProt P19404 subcellular location). NDUFV2 is assigned to the mitochondrial inner membrane and has no annotated transmembrane segment, so a crisp plasma-membrane outline is not its expected IHC pattern (UniProt P19404 topology). HPA reports high staining in colon glandular cells and cardiomyocytes, which can serve as tissue-based pattern references (HPA: High in colon glandular cells and cardiomyocytes). HPA IF also reports additional nucleoplasmic and cytosolic localisation; isolated nuclear DAB staining therefore needs independent validation before interpretation (HPA: approved IF locations; standard IHC practice). Compare cellular morphology, adjacent sections and negative detection controls when the compartment is ambiguous (standard IHC practice).
How should epitope uncertainty affect NDUFV2 IHC troubleshooting?
The supplied record lists 0 annotated isoforms and a processed chain spanning residues 33–249 (UniProt P19404 processing and isoforms). It also lists acetylation at residue 61 and phosphorylation at residue 193, but supplies no antibody epitope (UniProt P19404 modified residues; datasheet A05801-2: epitope not supplied). Do not attribute a staining difference to an isoform or either modification without epitope and validation data (UniProt P19404 isoforms and modified residues; standard IHC interpretation). If another antibody is available, compare antibodies raised against independently documented epitopes on adjacent sections with matched controls (standard IHC practice). Report epitope uncertainty when interpreting discordant compartment or intensity patterns (standard IHC practice).
How can IF help resolve an ambiguous NDUFV2 IHC pattern?
Treat IF as a separate assay: HPA reports mainly mitochondrial signal, with additional nucleoplasmic and cytosolic signal, in ICC/IF images from Hep-G2 and U2OS (HPA: approved subcellular locations and ICC/IF images). For tissue IF, multiplex NDUFV2 with a marker for the expected cell population and a mitochondrial marker, then assess signal overlap within identified cells (standard IF practice; UniProt P19404 subcellular location). Choose fluorophores after checking tissue autofluorescence in an unstained section and keep acquisition settings consistent (standard IF practice). Because NDUFV2 is at the inner mitochondrial membrane, optimise permeabilisation to give antibody access to the intracellular epitope once its location is known (UniProt P19404 topology; standard IF practice). The paraffin IHC caption provides no IF fixation or permeabilisation conditions (datasheet A05801-2).
What should I check when NDUFV2 DAB staining looks diffuse?
First compare the slide with a no-primary control and inspect whether colour appears in the expected granular cytoplasmic pattern (standard IHC practice; HPA: granular cytoplasmic profile). The selected paraffin example used 10% goat serum, 2 μg/ml primary antibody and DAB detection; these are useful starting conditions for that catalog antibody (datasheet A05801-2). For widespread DAB colour, check peroxidase blocking, antibody concentration, washes and development time on matched sections (standard IHC practice). Evaluate pigment and endogenous enzyme activity with appropriate detection controls before calling weak diffuse colour positive (standard IHC practice). Reassess the pattern in a positive reference tissue if background obscures individual cells (HPA: High in colon glandular cells; standard IHC practice).
How should I quantify NDUFV2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then record the percentage of positive cells and an intensity-based H-score where individual cells can be resolved (standard IHC practice). For a granular signal, a fixed-threshold DAB-positive area or optical-density measure can supplement cell scoring when segmentation is reproducible (HPA: granular cytoplasmic profile; standard IHC practice). Normalise to viable tissue area, or to the number of evaluable cells in the same population, and report the denominator (standard IHC practice). Keep section thickness, imaging settings, staining batch and thresholds consistent across comparisons (standard IHC practice). Exclude folds, edges and necrotic regions using prespecified criteria (standard IHC practice).
How do I distinguish true NDUFV2 positivity from IHC artefact?
A plausible positive pattern is granular cytoplasmic DAB staining in intact cells, consistent with mitochondrial NDUFV2 (HPA: granular cytoplasmic profile; UniProt P19404 subcellular location). Check whether an expected positive population, such as colon glandular cells, stains appropriately within the run (HPA: High in colon glandular cells; standard IHC practice). Surface outlining, edge-restricted colour, staining over necrosis or signal in the no-primary control warrants an artefact check (UniProt P19404 topology; standard IHC practice). Peroxidase-related colour can mimic weak positivity, so compare detection controls and tissue morphology before scoring (standard IHC practice). HPA reports only medium agreement between staining and RNA data; avoid treating a single IHC pattern as definitive expression evidence (HPA: Enhanced reliability description).
Boster reagents
Best NDUFV2 / NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial IHC Antibodies
A05801-2 has IHC images from human paraffin sections and an IF image from a human paraffin section (catalog image captions); listed reactivity includes human, mouse and rat (catalog reactivity).
Real IHC data
Anti-NDUFV2 Antibody ®
Cat # A05801-2
A05801-2 is the only listed SKU; its IHC images show human colorectal adenocarcinoma, esophageal squamous carcinoma and lung cancer paraffin sections (catalog IHC image captions). Its IF image shows a human rectal cancer paraffin section (catalog IF image caption).
Which to pick: Choose A05801-2 for tissue IHC: its paraffin section images used EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A05801-2 IHC image captions); the fixative is unreported (A05801-2 IHC image captions). For tissue IF, A05801-2 has a paraffin section image at 5 μg/ml (A05801-2 IF image caption); ICC is not listed among its applications (catalog applications). For mouse or rat samples, A05801-2 lists both species as reactive, while its supplied IHC and IF images show human sections (catalog reactivity; A05801-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P19404 (NDUV2_HUMAN, NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial).
- Human Protein Atlas. NDUFV2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. NDUFV2 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm and cytosol..
- Human Protein Atlas. NDUFV2 antibody validation summary (2 antibodies).
- Protein profiling identified mitochondrial dysfunction and synaptic abnormalities after dexamethasone intervention in rats with traumatic brain injury. Neural regeneration research 2021 — PMC8374556.
- Hypoxic tumor exosomes suppress macrophage inflammation and ferroptosis via NDUFV2 to enhance bystander tumor radioresistance. Cell death & disease 2025 — PMC12847814.
- MYC_V1-Related Genes Affect Gastric Cancer Proliferation by Regulating Energy Metabolism and Analysis of Therapeutic Targets. International journal of molecular sciences 2026 — PMC13256221.
- PubMed PMID:2500970 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:12611891 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NDUFV2 (NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial). Expected localisation Granular cytoplasmic staining (HPA tissue IHC); inner membrane at the molecular level (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05801-2). Validated IHC antibody: A05801-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet A05801-2); 2. Fixation: Image fixative and duration unreported (datasheet A05801-2); 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 A05801-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05801-2); 8. Primary antibody: Rabbit anti-NDUFV2, 2-5 μg/ml (datasheet A05801-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05801-2). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No isoforms annotated; mature chain begins at residue 33 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt P19404, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A05801-2, PMC8374556, PMC12847814. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NDUFV2/immunohistochemistry-ihc-guide.