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- Table of Contents
MT-ND6 / NADH-ubiquinone oxidoreductase chain 6 · Western blot design guide
Design a Western Blot for MT-ND6
Source-linked MT-ND6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MT-ND6 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-ND6 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 | ~18.6 kDa | |
| Observed band | ~19 kDa | |
| Gel | 5–20% (catalog A03777-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Regulation | Not found in UniProt | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MT-ND6 Western Blot Protocol Options
The A03777-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HepG2, human RT4 (catalog A03777-2) |
| Gel % | 5–20% (catalog A03777-2) |
| Load | 30 ug; reducing conditions (catalog A03777-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03777-2) |
| Membrane | nitrocellulose membrane (catalog A03777-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03777-2) |
| Primary antibody | A03777-2 · 0.5 μg/mL (catalog A03777-2) |
| Primary incubation | overnight at 4°C (catalog A03777-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03777-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03777-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03777-2) |
| Detection | ECL (catalog A03777-2) |
Section 2
What Is the Expected MT-ND6 Western Blot Band Size?
MT-ND6 is predicted at 18.6 kDa and observed at ~19 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~19 kDa | Matches the empirical MT-ND6 band in whole-cell lysates |
| Band near 18.6 kDa | Consistent with the predicted MT-ND6 mass; confirm its identity |
| Faint band near 19 kDa | May reflect limited recovery of this inner-membrane protein |
| No band near 19 kDa in whole-cell lysate | May reflect limited recovery or detection of this inner-membrane protein |
💡Expected MT-ND6 appearanceMT-ND6 has a predicted mass of 18.6 kDa and an empirical band at ~19 kDa in reducing whole-cell blots; confirm band identity with appropriate controls.
| UniProt predicted mass | Places full-length MT-ND6 at 18.6 kDa |
| Calculated full-length mass | Provides the reference size of 18.6 kDa |
| Predicted protein size | Supports a band near 19 kDa |
| UniProt molecular weight | Corresponds to 18.6 kDa for the listed sequence |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Inner-membrane MT-ND6 may be poorly recovered | Check mitochondrial enrichment and membrane-protein extraction |
| Weak or no signal | Limited recovery or transfer of this multi-pass membrane protein | Check extraction, transfer and a positive-control lysate |
| Band higher than expected | Identity and cause are not established by the listed features | Compare with the ~19 kDa reference band and verify antibody specificity |
| Band lower than expected | Identity and cause are not established by the listed features | Verify antibody specificity and check sample integrity |
| Multiple bands | The listed record supplies one isoform and does not explain extra bands | Use a positive control and an independent antibody to assess band identity |
Experiment evidence
Western blot images showing MT-ND6 band
A03777-2 · MT-ND6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MT-ND6 Western blot
🧪For positive controls for MT-ND6 in Western blot, you can use an HPA-validated tissue or cell sample once one is identified; none is supplied here.
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 data to establish a positive sample or a clean negative tissue.
HPA tissue expression evidence for MT-ND6
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-ND6 Western Blot Tips
Deeper troubleshooting and optimisation questions for MT-ND6, answered from its protein features.
How should MT-ND6 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 MT-ND6 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands; verify additional bands before assigning them to MT-ND6.
Could a listed modification explain an MT-ND6 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features do not support assigning a shifted band to a specific modification, and feature annotations alone would not establish a visible shift.
Does this guide establish induction of MT-ND6?
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-ND6 Western blot?
Transfer · MT-ND6 is a 174-residue, multi-pass inner-mitochondrial-membrane protein with a predicted mass of 18.6 kDa. Choose transfer conditions that retain proteins near 19 kDa, then check the membrane signal and post-transfer gel to assess recovery.
How should blocking be optimized for MT-ND6?
Blocking · Because MT-ND6 is a multi-pass membrane protein, compare blocking conditions using the same sample and antibody. Choose the condition that preserves the band near 19 kDa while reducing background.
How should MT-ND6 bands be quantified?
Quantitation · Quantify the consistently identified band near 19 kDa using exposures within the detection system’s linear range. Keep sample preparation and transfer conditions consistent across samples; MT-ND6 is an inner-mitochondrial-membrane protein, so compare equivalent mitochondrial material where possible.
Should MT-ND6 run at its predicted 18.6 kDa?
Interpretation · The reported band near 19 kDa is close to the predicted 18.6 kDa. Apparent migration can vary, so compare samples against a molecular-weight marker and a consistent reference sample.
MT-ND6 is a core subunit of the 45-subunit Complex I, but this annotation does not establish the identity of a higher-mass band. Check whether the band is reproducible and antibody-dependent before attributing it to an MT-ND6-containing complex.
MT-ND6 is a core subunit of the 45-subunit Complex I, but this annotation does not establish the identity of a higher-mass band. Check whether the band is reproducible and antibody-dependent before attributing it to an MT-ND6-containing complex.
Boster reagents
MT-ND6 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-ND6/MT-ND6 Antibody Picoband®
Cat # A03777-2
The catalog reports one anti-MT-ND6 antibody for Western blotting. Its image shows an approximately 19 kDa band in human HeLa, HepG2, and RT4 whole cell lysates. The supplied evidence does not establish performance in other specimens or independent target validation.
Which to pick: A03777-2 is the only listed option. It reports human reactivity and includes a Western blot image from HeLa, HepG2, and RT4 lysates, with a band near the expected 19 kDa size. For other sample types, validation is not shown.
Source: BosterBio MT-ND6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P03923.
- Human Protein Atlas. MT-ND6 tissue expression.
- PMC7719573 — target-verified WB comparison
- PMC13331803 — target-verified WB comparison
- PMC10399063 — target-verified WB comparison