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- Table of Contents
NDUFV2 / NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial · Western blot design guide
Design a Western Blot for NDUFV2
Source-linked NDUFV2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NDUFV2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
NDUFV2 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~27.4 kDa | |
| Observed band | ~27 kDa | |
| Gel | 5–20% (catalog A05801-2) | |
| Positive control | Bronchus (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked NDUFV2 Western Blot Protocol Options
The A05801-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human A549, human HepG2 (catalog A05801-2) |
| Gel % | 5–20% (catalog A05801-2) |
| Load | 30 ug; reducing conditions (catalog A05801-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05801-2) |
| Membrane | nitrocellulose membrane (catalog A05801-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05801-2) |
| Primary antibody | A05801-2 · 0.25 μg/mL (catalog A05801-2) |
| Primary incubation | overnight at 4°C (catalog A05801-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05801-2) |
| Secondary incubation | 1.5 hour at RT (catalog A05801-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05801-2) |
| Detection | ECL (catalog A05801-2) |
Section 2
What Is the Expected NDUFV2 Western Blot Band Size?
NDUFV2 is predicted at 27.4 kDa and observed at ~27 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~27 kDa | Matches the empirical NDUFV2 band and its 27.4 kDa predicted mass |
| Weak ~27 kDa band in whole-cell lysate | NDUFV2 is a mitochondrial inner-membrane protein whose recovery may vary |
| ~27 kDa band in cell lysates | Consistent with the reported band across multiple cell lines |
| ~27 kDa band in tissue lysates | Consistent with the reported band in heart, brain, and kidney samples |
💡Expected NDUFV2 appearanceNDUFV2 is predicted at 27.4 kDa and has an empirical band at ~27 kDa; confirm band identity with appropriate antibody and sample controls.
| UniProt predicted mass | Places the expected protein near 27.4 kDa, consistent with the ~27 kDa observed band |
| Lys61 acetylation | Is annotated, but no visible migration effect is established |
| Tyr193 phosphorylation by SRC | Is annotated, but no visible migration effect is established |
| Transit peptide annotation | Does not establish a cleavage site or a separate mature-band size |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Limited recovery of mitochondrial NDUFV2 | Check mitochondrial protein recovery and use a positive lysate control |
| Band higher than expected | Band identity or migration is uncertain | Compare with a validated positive sample and an independent antibody |
| Band lower than expected | Band identity or protein integrity is uncertain | Check sample integrity and confirm with an independent antibody |
| Multiple bands | Additional bands are not explained by the single listed isoform | Compare positive and negative controls and validate antibody specificity |
| Weak or no signal | Low recovery or detection of mitochondrial NDUFV2 | Check loading and transfer near 27 kDa and enrich mitochondria if needed |
| Fragments below expected size | Protein degradation or nonspecific recognition is possible | Check sample handling and confirm the ~27 kDa band with an independent antibody |
Experiment evidence
Western blot images showing NDUFV2 band
A05801-2 · NDUFV2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NDUFV2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NDUFV2 in Western blot, you can use bronchus tissue, which shows high HPA expression.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Mitochondrial enrichment may improve detection of this inner-membrane protein.
HPA tissue expression evidence for NDUFV2
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for NDUFV2, answered from its protein features.
How should NDUFV2 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could NDUFV2 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence. Do not assign additional bands to NDUFV2 isoforms from these features alone.
Which NDUFV2 modifications matter when interpreting bands?
PTM · UniProt lists N6-acetyllysine at position 61 and phosphotyrosine at position 193. These are UniProt coordinates; antibody or paper numbering may differ. Neither listed modification establishes a visible band shift.
UniProt assigns phosphotyrosine at position 193 to SRC. A phosphotyrosine-sensitive assay may assess that site, but the feature alone does not establish its occupancy or predict a change in total NDUFV2 signal.
UniProt assigns phosphotyrosine at position 193 to SRC. A phosphotyrosine-sensitive assay may assess that site, but the feature alone does not establish its occupancy or predict a change in total NDUFV2 signal.
Does this guide establish induction of NDUFV2?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for NDUFV2 Western blot?
Transfer · NDUFV2 is a roughly 27 kDa peripheral protein associated with the mitochondrial inner membrane. Choose transfer conditions suitable for a protein of that size and check transfer efficiency; the listed features do not specify a required method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05801-2 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should NDUFV2 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why is the NDUFV2 band near 27 kDa?
Interpretation · The observed band near 27 kDa is close to the predicted 27.4 kDa. The listed features do not establish a cause for the small difference in apparent mass.
NDUFV2 is one core subunit of the 45-subunit Complex I. Quantify its band consistently across samples, but interpret the signal as NDUFV2 abundance rather than a direct measure of Complex I activity or assembly.
Check whether the expected band near 27 kDa is present and verify additional bands before assigning them to NDUFV2. The listed single isoform, two modification sites, and absence of listed glycosylation sites do not by themselves explain extra bands.
NDUFV2 is one core subunit of the 45-subunit Complex I. Quantify its band consistently across samples, but interpret the signal as NDUFV2 abundance rather than a direct measure of Complex I activity or assembly.
Check whether the expected band near 27 kDa is present and verify additional bands before assigning them to NDUFV2. The listed single isoform, two modification sites, and absence of listed glycosylation sites do not by themselves explain extra bands.
Boster reagents
NDUFV2 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-NDUFV2 Antibody Picoband®
Cat # A05801-2
The catalog reports one anti-NDUFV2 antibody, A05801-2, with reported human, mouse, and rat reactivity. Its WB image shows an approximately 27 kDa band in the named cell and tissue lysates under reducing conditions. No independent specificity validation is supplied.
Which to pick: A05801-2 is the only listed option and has a WB image. Match your sample to the caption’s tested human cell lines or rat and mouse cells and tissues; the documented primary antibody concentration was 0.25 μg/mL.
Source: BosterBio NDUFV2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P19404.
- Human Protein Atlas. NDUFV2 tissue expression.
- PMC8613759 — target-verified WB comparison
- PMC3117770 — target-verified WB comparison
- PMC10366351 — target-verified WB comparison