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NDUFB2 / NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mitochondrial · IHC design guide
Design Immunohistochemistry for NDUFB2
Plan chromogenic NDUFB2 IHC in paraffin sections using the IHC-validated antibody A13719-1 (datasheet). Compare staining with the cytoplasmic and nuclear tissue pattern (HPA tissue IHC), while assessing its fit with the mitochondrial inner-membrane assignment (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NDUFB2 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 and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | General cytoplasmic and nuclear staining; nucleoli in a few tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A13719-1) | |
| Positive control | Ovary+4 more · see all | |
| Negative control | Bone marrow+1 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 | Nuclear staining differs from the inner-membrane assignment (HPA tissue IHC; UniProt) | |
| Regulation | Low tissue RNA specificity (HPA tissue RNA) | |
| Isoform / epitope | No isoforms annotated; map epitopes to processed chain 34–105 (UniProt) |
Section 1
Recommended NDUFB2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: EDTA pH 8.0 HIER) is accompanied by 1 published glioblastoma IHC protocol (PMC12777021).
| Sample | Paraffin-embedded human liver tissue; fixative not specified (datasheet A13719-1) |
| Fixation | Image fixative and duration unreported (datasheet A13719-1); 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 A13719-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A13719-1) |
| Primary antibody | Rabbit anti-NDUFB2, 2-5μg/ml (datasheet A13719-1) |
| Primary incubation | Overnight at 4 °C (datasheet A13719-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A13719-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NDUFB2-positive staining in follicle cells of ovary (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and nuclear expression with additional nucleoli in a few tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet); the published protocol specifies microwave retrieval without a buffer (PMC12777021).
Section 2
What Is the Expected NDUFB2 Staining Pattern?
NDUFB2 is an inner mitochondrial membrane Complex I subunit with no annotated transmembrane segment (UniProt O95178 topology). In paraffin-section IHC, expect staining in ovarian follicle cells and cells in testicular seminiferous ducts, where HPA reports high signal (HPA: High). HPA also describes general cytoplasmic and nuclear staining. Its tissue IHC reliability is Approved, with medium agreement with RNA data and external verification pending (HPA: reliability).
| Follicle cells or cells in seminiferous ducts stain strongly, with cytoplasmic signal (HPA: High in both). | This fits the strongest supplied tissue observations and the mitochondrial location (HPA: tissue IHC; UniProt O95178). Judge cell identity and compartment together: a strong brown deposit elsewhere in the section alone does not reproduce the reported positive pattern (HPA: tissue IHC). |
| Nuclear staining accompanies cytoplasmic staining, sometimes with nucleoli (HPA: tissue IHC profile). | HPA reports this pattern in tissue IHC, so nuclear signal alone cannot be called an artefact (HPA: tissue IHC profile). Interpret isolated, dominant nuclear staining cautiously: UniProt places NDUFB2 at the mitochondrial inner membrane, and HPA rates nucleoplasmic localization uncertain in ICC-IF (UniProt O95178; HPA: subcellular). |
| An unexpected cell population stains more strongly than the reported cells (HPA: tissue IHC). | First check the section’s cell identification and the matched negative control (general IHC practice). If the pattern persists, cross-reactivity or endogenous detection activity is possible (general IHC practice). HPA reports low tissue specificity, so staining outside the listed high-signal cells is not, by itself, proof of either problem (HPA: RNA specificity). |
| Brown colour spreads across tissue, stroma, or the whole section without clear cellular boundaries. | Treat this as background rather than a scored cellular pattern (general IHC practice). Compare a matched negative control, then review blocking, washes, detection chemistry, and section condition (general IHC practice). The supplied HPA observations describe cellular staining; they do not establish a target-specific cause for diffuse colour (HPA: tissue IHC). |
| A verified ovary or testis section shows no signal in the listed high-signal cells (HPA: High). | A failed positive control makes a negative result in a test specimen hard to interpret (general IHC practice). Confirm the expected cells are present, then check the validated antibody, reagent activity, retrieval, and detection run (general IHC practice). HPA’s Approved rating still carries medium RNA agreement and pending external verification (HPA: reliability). |
💡Expected NDUFB2 appearanceCall an IHC result consistent when ovarian follicle cells or cells in seminiferous ducts show clear, high cellular signal, including cytoplasmic staining (HPA: High; HPA: tissue IHC profile); diffuse acellular colour or control-matched signal is suspect (general IHC practice).
| Tissue and cell choice (HPA: tissue IHC) | Ovarian follicle cells and cells in seminiferous ducts are the supplied high-signal examples; adrenal and appendix glandular cells are medium, while bone marrow hematopoietic cells are not detected (HPA: tissue IHC). Match the comparison to the named cell population, not merely the organ (general IHC practice). |
| Compartment evidence (UniProt O95178; HPA: subcellular) | UniProt assigns NDUFB2 to the mitochondrial inner membrane and annotates no transmembrane segment (UniProt O95178 topology). HPA tissue IHC reports cytoplasmic and nuclear staining; ICC-IF adds approved mitochondrial localization but uncertain nucleoplasmic localization (HPA: tissue IHC; HPA: subcellular). Keep these evidence types distinct. |
| Antibody evidence (HPA: antibodies and reliability) | The listed rabbit polyclonal antibody, HPA051377, is Approved for IHC and ICC (HPA: antibodies). The tissue IHC assessment reports medium consistency with RNA expression and awaits external verification (HPA: reliability). An Approved label supports use of the observed pattern but does not settle every unexpected compartment or cell type. |
| Processing and epitope limits (UniProt O95178) | UniProt lists a chain spanning residues 34–105 and no annotated glycosylation sites, modified residues, or isoforms (UniProt O95178). The payload gives no antibody epitope or NDUFB2-specific retrieval response; it cannot predict which retrieval condition will recover staining or how fixation will affect this target. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in the named high-signal cells of ovary or testis (HPA: High). | The run may have failed, or the expected cells may be absent from the examined section (general IHC practice). | Confirm cell identity and section quality; compare a known-positive section from the same run, then check retrieval and detection reagents (general IHC practice). Avoid scoring other specimens as NDUFB2-negative until the positive control works. |
| Broad, even brown staining obscures cell boundaries. | Background from blocking, washes, or detection chemistry is possible (general IHC practice). | Inspect the matched negative control and review blocking, wash, and detection steps (general IHC practice). Score only resolvable cellular staining; the HPA tissue profile describes cellular compartments, not diffuse section-wide colour (HPA: tissue IHC profile). |
| A supposedly negative comparison stains strongly. | The compared cells may differ from the HPA-listed population; cross-reactivity or endogenous detection activity is also possible (HPA: tissue IHC; general IHC practice). | Verify cell identity first. HPA reports bone marrow hematopoietic cells and parathyroid glandular cells as not detected, but does not establish every cell in those organs as negative (HPA: tissue IHC). Check a matched negative control if the discrepancy remains. |
| Staining is predominantly nuclear or nucleolar. | Some nuclear and occasional nucleolar tissue staining is reported, while nucleoplasmic ICC-IF localization is uncertain (HPA: tissue IHC profile; HPA: subcellular). | Document the compartment and compare cytoplasmic signal, expected cell types, and the negative control (HPA: tissue IHC; general IHC practice). Do not call nuclear signal alone definitive mitochondrial NDUFB2 localization (UniProt O95178; HPA: subcellular). |
| An unexpected cell type is the strongest stained population. | Cell identification, cross-reactivity, or endogenous detection activity may explain a mismatch (general IHC practice). HPA reports low tissue RNA specificity, so expression outside listed examples remains possible (HPA: RNA specificity). | Recheck morphology and compare the matched negative control (general IHC practice). Report the unexpected population separately from the HPA high-signal examples rather than assigning it the same expected intensity (HPA: tissue IHC). |
| Q: Does nuclear staining in ICC-IF resolve an IHC localization question? | A: HPA labels nucleoplasm uncertain and additional mitochondrial localization approved in ICC-IF; tissue IHC reports nuclear and cytoplasmic staining (HPA: subcellular; HPA: tissue IHC profile). | Use the ICC-IF observations as context, not a substitute for interpreting the paraffin section (HPA: subcellular; general IHC practice). Compare IHC cell type, compartment, and controls on its own terms; ICC-IF workflow belongs in its separate guide. |
Experiment evidence
IHC & IF images showing NDUFB2 staining
A13719-1 · IHCIHC analysis of NDUFB2 using anti-NDUFB2 antibody (A13719-1). NDUFB2 was detected in paraffin-embedded section of human liver tissue. Heat m
A13719-1 · IHCIHC analysis of NDUFB2 using anti-NDUFB2 antibody (A13719-1). NDUFB2 was detected in paraffin-embedded section of human lung cancer tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000090266-NDUFB2/tissue/cerebral+cortex#imid_15122706
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000090266-NDUFB2/tissue/colon#imid_15122616
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000090266-NDUFB2/tissue/liver#imid_15122633
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000090266-NDUFB2/tissue/kidney#imid_15122634
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA051377; green = NDUFB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA051377; green = NDUFB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NDUFB2 IHC & IF
🧪Run ovary first and score staining in follicle cells (HPA: ovary follicle cells High). Use bone marrow hematopoietic cells as the negative tissue (HPA: bone marrow hematopoietic cells Not detected); on the ovary slide, compare adjacent non-follicle cells, but count them as an internal negative only if they lack staining, because no negative ovarian cell type is identified in the supplied HPA rows (HPA: ovary follicle cells High).
Positive control tissue: Ovary (Follicle cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NDUFB2 in A-549, MCF-7, with annotated localisation: Nucleoplasm (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched rabbit IgG isotype control (caption: rabbit primary antibody; standard IHC practice); use matched NDUFB2 knockout material as a biological negative if available (standard IHC practice). For the reported biotin-based DAB workflow, block endogenous peroxidase and check for endogenous biotin background in the ovary section (caption: biotinylated secondary, streptavidin-biotin complex and DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A13719-1 tissue-IHC caption does not state a fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 preceded staining in the reported paraffin-section example, but a retrieval requirement or advantage for frozen sections or IF has not been established by the supplied evidence (caption: EDTA retrieval in a paraffin section). With the reported biotin-based DAB detection, assess ovarian background in the controls before scoring follicle-cell staining (caption: biotinylated secondary, streptavidin-biotin complex and DAB; standard IHC practice).
HPA tissue IHC evidence for NDUFB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Ovary | Follicle cells | High | Protein (IHC) | HPA → |
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced NDUFB2 IHC Tips
Troubleshoot NDUFB2 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting signal.
Which retrieval condition should I start with for NDUFB2 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A13719-1). The selected paraffin-section image used that condition before 10% goat-serum blocking and 2 µg/mL primary antibody overnight at 4°C (datasheet A13719-1). If staining is weak, adjust heating time or cooling consistently across a pilot set, then compare signal and tissue preservation using the same detection conditions (standard IHC practice). Record the retrieval device, temperature and duration for each run; the caption does not specify those settings, so a reproducible local setting must be established (datasheet A13719-1; standard IHC practice).
How should I troubleshoot fixation-related loss of NDUFB2 staining?
The selected image documents a paraffin-embedded human liver section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A13719-1). Compare sections with documented fixation and processing histories while holding EDTA pH 8.0 retrieval and antibody incubation constant (datasheet A13719-1; standard IHC practice). Inspect morphology alongside staining: poor preservation can make a change in signal difficult to interpret (standard IHC practice). Do not use the HPA tissue pattern or NDUFB2 inner-membrane annotation to infer a preferred fixative or fixation duration; neither supplies a target-specific fixation comparison (HPA tissue IHC; UniProt O95178).
Should NDUFB2 appear mitochondrial or nuclear in chromogenic IHC?
NDUFB2 is annotated at the mitochondrial inner membrane, so assess cytoplasmic staining for a plausible mitochondrial distribution (UniProt O95178 topology). HPA tissue IHC reports general cytoplasmic and nuclear staining, with nucleoli in a few tissues, while its subcellular data mark nucleoplasm as uncertain and mitochondria as approved (HPA tissue IHC; HPA subcellular). Nuclear signal therefore needs a specificity check rather than automatic acceptance as NDUFB2 (UniProt O95178; HPA subcellular; standard IHC practice). Compare cell morphology, adjacent structures and a no-primary control under matched DAB development; diffuse nuclear or section-edge staining that tracks the control warrants investigation (standard IHC practice).
Could an isoform or epitope explain inconsistent NDUFB2 staining?
The supplied record lists no annotated isoforms, glycosylation sites or modified residues for NDUFB2 (UniProt O95178). It annotates a chain spanning residues 34–105 and no transmembrane segment, but the selected antibody caption does not map its epitope (UniProt O95178; datasheet A13719-1). Do not attribute a staining difference to an isoform or a particular modification without independent evidence (UniProt O95178; standard IHC practice). Instead, compare retrieval, section processing and antibody incubation in matched sections, then seek an independent antibody with a documented epitope if specificity remains uncertain (standard IHC practice).
How can IF help check an ambiguous NDUFB2 IHC pattern?
Use IF as a separate localisation check, multiplexing NDUFB2 with a mitochondrial marker and, where cell identity is disputed, a marker of the expected cell type (UniProt O95178; standard IF practice). Choose spectrally separated fluorophores and a channel with low tissue autofluorescence, checking an unstained section in each channel (standard IF practice). NDUFB2 is assigned to the inner membrane, but its antibody epitope and membrane-facing side are unspecified; titrate permeabilisation against mitochondrial morphology rather than assuming a particular side is accessible (UniProt O95178; datasheet A13719-1; standard IF practice). Compare the resulting distribution with the chromogenic pattern without transferring the paraffin-section retrieval conditions to IF (datasheet A13719-1; standard IF practice).
What should I check when DAB background obscures NDUFB2?
Run a no-primary section through the same secondary and DAB steps to locate detection-related background (standard IHC practice). The selected image used 10% goat serum, biotinylated goat anti-rabbit IgG, a streptavidin–biotin complex and DAB; keep those steps consistent while isolating a cause (datasheet A13719-1). Check peroxidase blocking and, with this detection format, endogenous biotin as possible contributors; these are general workflow controls, not evidence of NDUFB2 expression (datasheet A13719-1; standard IHC practice). Also compare wash stringency and DAB development time on matched sections, because excess development can mask a cytoplasmic pattern (standard IHC practice).
How should I quantify NDUFB2 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell populations and tissue regions before scoring, then use an H-score or the percentage of positive cells with intensity categories applied consistently (standard IHC practice). Normalize counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when area is the denominator (standard IHC practice). Record cytoplasmic and nuclear staining separately because HPA reports both, while the mitochondrial inner membrane is the UniProt localisation (HPA tissue IHC; UniProt O95178). Keep retrieval, antibody incubation, DAB development, imaging and thresholds matched across samples; include control sections to detect run-to-run drift (datasheet A13719-1; standard IHC practice).
How do I distinguish convincing NDUFB2 staining from artefact?
Favor reproducible cytoplasmic staining with plausible mitochondrial distribution in intact cells, consistent with the inner-membrane annotation (UniProt O95178; standard IHC practice). HPA reports high staining in ovarian follicle cells and cells in seminiferous ducts, but low staining in hepatocytes; its tissue-IHC reliability is Approved with medium RNA concordance and pending external verification (HPA tissue IHC). Treat isolated nuclear signal cautiously because HPA labels nucleoplasm uncertain while mitochondrial localisation is approved (HPA subcellular). Exclude section-edge staining, necrotic areas and signal reproduced by a no-primary control; check endogenous peroxidase and biotin when interpreting this DAB and streptavidin–biotin workflow (datasheet A13719-1; standard IHC practice).
Boster reagents
Best NDUFB2 / NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mitochondrial IHC Antibodies
A13719-1 has real IHC images from paraffin-embedded human liver, lung cancer, and rectal cancer sections; no IF image is supplied (catalog image captions and IF image list).
Real IHC data
Anti-NDUFB2 Antibody ®
Cat # A13719-1
A13719-1 is listed for human IHC and shown on paraffin-embedded human liver sections (catalog applications, reactivity, and image captions). Its other IHC images show human lung cancer and rectal cancer sections (catalog image captions).
Which to pick: For tissue IHC, choose A13719-1; the catalog specifies 2–5 μg/ml for human paraffin sections and identifies the antibody as rabbit-hosted, with clonality unspecified (catalog dilution, reactivity, host, and clone fields). Its image captions describe heat retrieval in EDTA at pH 8.0, while the fixative is unreported (catalog image captions). No IF/ICC or cross-species SKU can be recommended from this catalog: A13719-1 has no listed IF/ICC application or IF image and lists human reactivity only (catalog applications, IF image list, and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95178 (NDUB2_HUMAN, NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mitochondrial).
- Human Protein Atlas. NDUFB2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NDUFB2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the mitochondria..
- Human Protein Atlas. NDUFB2 antibody validation summary (1 antibodies).
- Identifying NDUFB2 as a prognostic biomarker for glioblastoma through an exploratory analysis of an anesthesia-related gene signature. BMC cancer 2025 — PMC12777021.
- PubMed PMID:9878551 — UniProt-cited evidence.
- PubMed PMID:11042152 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NDUFB2 (NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mitochondrial). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A13719-1). Validated IHC antibody: A13719-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Ovary (Follicle cells); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver tissue; fixative not specified (datasheet A13719-1); 2. Fixation: Image fixative and duration unreported (datasheet A13719-1); 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 A13719-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A13719-1); 8. Primary antibody: Rabbit anti-NDUFB2, 2-5μg/ml (datasheet A13719-1); 9. Primary incubation: Overnight at 4 °C (datasheet A13719-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Nuclear staining differs from the inner-membrane assignment (HPA tissue IHC; UniProt); Isoform / epitope: No isoforms annotated; map epitopes to processed chain 34–105 (UniProt); Regulation: Low tissue RNA specificity (HPA tissue RNA). Evidence: UniProt O95178, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A13719-1, PMC12777021. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NDUFB2/immunohistochemistry-ihc-guide.