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MT-ND1 / NADH-ubiquinone oxidoreductase chain 1 · Western blot design guide
Design a Western Blot for MT-ND1
Real validated MT-ND1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MT-ND1 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-ND1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~35.7 kDa | |
| Observed band | ~36 kDa | |
| Gel | 12% (catalog A02292-1) | |
| Positive control | Heart muscle (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 | — | |
| Caveat | Membrane extraction controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MT-ND1 Western Blot Protocols
The A02292-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, rat heart, mouse brain (catalog A02292-1) |
| Gel % | 12% (catalog A02292-1) |
| Load | 30 ug; reducing conditions (catalog A02292-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02292-1) |
| Membrane | nitrocellulose membrane (catalog A02292-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02292-1) |
| Primary antibody | A02292-1 · 1:1000 (catalog A02292-1) |
| Primary incubation | overnight at 4°C (catalog A02292-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02292-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02292-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02292-1) |
| Detection | ECL (catalog A02292-1) |
Section 2
What Is the Expected MT-ND1 Western Blot Band Size?
MT-ND1 is predicted at 35.7 kDa and observed at ~36 kDa; the cause of the small difference is not established.
| Band at ~36 kDa | Matches the observed MT-ND1 band near its 35.7 kDa predicted mass |
| Faint band at ~36 kDa | May reflect limited recovery of this inner membrane protein |
| No band near ~36 kDa | May reflect inadequate recovery or detection of MT-ND1 |
| Additional band away from ~36 kDa | Its identity is not established by the listed MT-ND1 features |
💡Expected MT-ND1 appearanceMT-ND1 has a predicted mass of 35.7 kDa and an observed band at ~36 kDa; confirm unexpected bands with ordinary band-identity controls.
| UniProt predicted molecular mass | Sets the full-length reference at 35.7 kDa, close to the observed ~36 kDa band |
| Predicted mass in daltons | 35,661 Da corresponds to the same approximately 35.7 kDa reference |
| Predicted mass as a higher-band reference | Bands substantially above 35.7 kDa need separate identity confirmation |
| Predicted mass as a lower-band reference | Bands substantially below 35.7 kDa need separate identity confirmation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MT-ND1 is an inner membrane protein that may be poorly recovered | Check mitochondrial recovery and membrane solubilization |
| Weak or no signal | Limited recovery or detection of MT-ND1 | Check mitochondrial enrichment and antibody performance |
| Band higher than expected | The listed features do not establish a higher-mass MT-ND1 form | Check solubilization and confirm identity with an independent antibody or genetic depletion |
| Band lower than expected | No listed cleavage feature explains a smaller MT-ND1 band | Check sample integrity and confirm identity with genetic depletion |
| Multiple bands | The listed features do not establish multiple MT-ND1 forms | Confirm band specificity with an independent antibody or genetic depletion |
Experiment evidence
Western blot images showing MT-ND1 band
A02292-1 · MT-ND1
M02292 · MT-ND1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MT-ND1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MT-ND1 in Western blot, you can use heart muscle lysate.
Positive control: Heart muscle (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: As an inner mitochondrial membrane protein, MT-ND1 may require effective membrane protein extraction for a clear signal.
HPA tissue expression evidence for MT-ND1
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 |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | 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 → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MT-ND1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MT-ND1, answered from its protein features.
How should MT-ND1 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-ND1 isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning multiple bands to isoforms.
Do the listed modifications explain an MT-ND1 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not support assigning a shifted band to either modification. They also do not establish that any feature causes a visible shift.
Does this guide establish induction of MT-ND1?
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-ND1 Western blot?
Transfer · The supplied features identify MT-ND1 as a multi-pass inner-membrane protein but do not specify a transfer method. Check recovery and detection of the approximately 36 kDa band when evaluating transfer conditions.
How should blocking be optimized for MT-ND1?
Blocking · MT-ND1 is a multi-pass protein of the mitochondrial inner membrane. Compare blocking conditions using the same sample preparation and assess signal near 36 kDa alongside background. The supplied features do not identify a preferred blocker.
How should MT-ND1 bands be quantified?
Quantitation · Quantify the approximately 36 kDa band under consistent sample preparation, loading, transfer, and detection conditions. Because MT-ND1 is an inner-membrane protein and a Complex I core subunit, keep membrane extraction conditions consistent across samples.
Why is the MT-ND1 band near 36 kDa?
Interpretation · The observed band at approximately 36 kDa is close to the predicted 35.7 kDa for the 318-residue protein. The supplied features do not establish a cause for any small difference between apparent and predicted mass.
The record lists one isoform, no alternative sequence, and no signal peptide or propeptide. Those features do not explain additional bands. Compare any unexpected band with the approximately 36 kDa observed band before assigning it to MT-ND1.
The record lists one isoform, no alternative sequence, and no signal peptide or propeptide. Those features do not explain additional bands. Compare any unexpected band with the approximately 36 kDa observed band before assigning it to MT-ND1.
Boster reagents
MT-ND1 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-ND1/MT-ND1 Antibody
Cat # A02292-1
Real WB data
Anti-MT-ND1 Rabbit Monoclonal Antibody
Cat # M02292
Two the supplier anti-MT-ND1 antibodies have WB images. A02292-1 shows an approximately 36 kDa band in rat and mouse brain and heart lysates. M02292 shows WB analysis of human fetal muscle lysate. These images document the reported samples, not broader validation.
Which to pick: For human fetal muscle, M02292 has the matching WB image and lists human, mouse, and rat reactivity. For mouse brain or heart, A02292-1 has matching images. Its caption also shows rat tissues, though its listed reactivity is human and mouse.
Source: BosterBio MT-ND1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P03886.
- Human Protein Atlas. MT-ND1 tissue expression.
- PMC6861238 — target-verified WB comparison
- PMC5877625 — target-verified WB comparison
- PMC8322717 — target-verified WB comparison