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- Table of Contents
MT-ND2 / NADH-ubiquinone oxidoreductase chain 2 · Western blot design guide
Design a Western Blot for MT-ND2
Source-linked MT-ND2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MT-ND2 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
MT-ND2 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 | ~39 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Membrane extraction controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MT-ND2 Western Blot Protocol Options
The A04475 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | The brain of mouse(30ug), The brain of Rat(30ug) (catalog A04475) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A04475; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected MT-ND2 Western Blot Band Size?
MT-ND2 is predicted at 39 kDa; inner-membrane localization may affect recovery, but a migration difference has not been demonstrated.
| Band near 39 kDa | Consistent with the predicted MT-ND2 mass; confirm its identity with controls |
| Weak band near 39 kDa in whole-cell lysate | Limited recovery of inner-membrane MT-ND2 is possible |
| No band in whole-cell lysate | Mitochondrial membrane protein extraction may be insufficient |
| Stronger band near 39 kDa after mitochondrial enrichment | Consistent with MT-ND2 localization in the mitochondrial inner membrane |
💡Expected MT-ND2 appearanceMT-ND2 is predicted to be 39 kDa, with no empirical band size supplied; evaluate a band near that size using mitochondrial enrichment and band-identity controls.
| UniProt predicted mass | Provides a 39 kDa reference for full-length MT-ND2 |
| Calculated molecular weight | 38,961 Da rounds to approximately 39 kDa |
| Predicted full-length size | Does not establish the apparent band position on a blot |
| Sequence-based mass estimate | Provides no evidence for a distinct shifted band |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of mitochondrial inner-membrane MT-ND2 | Check mitochondrial enrichment and membrane protein extraction |
| Band higher than expected | Band identity or apparent migration is uncertain | Compare mitochondrial-enriched and control samples |
| Band lower than expected | Possible degradation or unrelated antibody signal | Check sample integrity and band-identity controls |
| Multiple bands | No supplied MT-ND2 feature establishes multiple band sizes | Compare bands across mitochondrial-enriched and control samples |
| Weak or no signal | Inefficient recovery of a multi-pass inner-membrane protein | Assess membrane extraction and sample loading |
Experiment evidence
Western blot images showing MT-ND2 band
A04475 · MT-ND2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MT-ND2 Western blot
🧪For positive controls for MT-ND2 in Western blot, you can use a sample containing mitochondrial inner membranes, but the supplied HPA data identify no positive tissue or cell line.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no tissue-expression data, so a positive tissue and a clean negative tissue cannot be established from this evidence.
HPA tissue expression evidence for MT-ND2
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced MT-ND2 Western Blot Tips
Deeper troubleshooting and optimisation questions for MT-ND2, answered from its protein features.
How should MT-ND2 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 an MT-ND2 isoform explain a second band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for a second band. Check band specificity before assigning either band to MT-ND2.
Could annotated modifications explain an MT-ND2 band shift?
PTM · The supplied features list no modified residues or glycosylation sites, and no signal peptide or propeptide. They provide no annotated modification or processing event to assign to a shifted band. They also cannot rule out an unlisted cause.
Does this guide establish induction of MT-ND2?
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 MT-ND2 Western blot?
Transfer · MT-ND2 is a predicted 39-kDa, multi-pass mitochondrial inner-membrane protein. Select transfer conditions appropriate for that size and verify transfer by checking both the membrane and post-transfer gel. The supplied features do not identify one optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04475 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 MT-ND2 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.
Should MT-ND2 run exactly at its predicted 39 kDa?
Interpretation · The supplied predicted mass is 39 kDa, but no observed band position is available. Use 39 kDa as a reference, not an expected exact migration position. The listed features alone cannot establish a visible shift or explain a difference between apparent and predicted mass.
MT-ND2 is a mitochondrial inner-membrane subunit of Complex I. Compare samples using consistent mitochondrial preparation and protein loading, and quantify the same validated band in each lane. A change in MT-ND2 signal alone does not establish a change in the entire 45-subunit complex.
First assess band specificity and sample preparation. The supplied record predicts 39 kDa, lists one isoform, and provides no modified residues, glycosylation sites, signal peptide, or propeptide. Those features do not assign an identity to an unexpected band or establish why it migrates differently.
MT-ND2 is a mitochondrial inner-membrane subunit of Complex I. Compare samples using consistent mitochondrial preparation and protein loading, and quantify the same validated band in each lane. A change in MT-ND2 signal alone does not establish a change in the entire 45-subunit complex.
First assess band specificity and sample preparation. The supplied record predicts 39 kDa, lists one isoform, and provides no modified residues, glycosylation sites, signal peptide, or propeptide. Those features do not assign an identity to an unexpected band or establish why it migrates differently.
Boster reagents
MT-ND2 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-MT-ND2 Antibody
Cat # A04475
A04475 is an anti-MT-ND2 polyclonal antibody listed for human, mouse, and rat. Its WB image shows mouse and rat brain tissue lysates (30 µg per lane) at 1:1000 dilution; the supplied image does not show a human sample.
Which to pick: A04475 is the only listed option. Its WB image provides a starting reference for mouse or rat brain lysate; human reactivity is listed, but no human WB sample is shown.
Source: BosterBio MT-ND2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P03891.
- Human Protein Atlas. MT-ND2 tissue expression.
- PMC13350946 — target-verified WB comparison
- PMC8120465 — target-verified WB comparison
- PMC7170900 — target-verified WB comparison