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NDUFA10 / NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 10, mitochondrial · Western blot design guide
Design a Western Blot for NDUFA10
Source-linked NDUFA10 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NDUFA10 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
NDUFA10 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 | ~40.8 kDa | |
| Observed band | ~41 kDa | |
| Gel | 10% (catalog A10517-1) | |
| Positive control | Adrenal gland (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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked NDUFA10 Western Blot Protocol Options
The A10517-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human RT4, human HepG2, human MCF-7, human PC-3 (catalog A10517-1) |
| Gel % | 10% (catalog A10517-1) |
| Load | 30 ug; reducing conditions (catalog A10517-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A10517-1) |
| Membrane | nitrocellulose membrane (catalog A10517-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A10517-1) |
| Primary antibody | A10517-1 · 0.5 μg/mL (catalog A10517-1) |
| Primary incubation | overnight at 4°C (catalog A10517-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A10517-1) |
| Secondary incubation | 1.5 hour at RT (catalog A10517-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A10517-1) |
| Detection | ECL (catalog A10517-1) |
Section 2
What Is the Expected NDUFA10 Western Blot Band Size?
NDUFA10 is predicted at 40.8 kDa and observed at ~41 kDa; isoforms and modifications are documented, but their effects on migration are not established.
| Band at ~41 kDa | Matches the observed NDUFA10 band and its 40.8 kDa predicted mass; confirm identity with controls. |
| ~41 kDa band in a mitochondrial fraction | Consistent with NDUFA10 localization to the mitochondrial matrix. |
| Several bands at different sizes | Isoforms 1 and 2 are possible contributors, but distinct migration is unverified. |
| Band shifted from ~41 kDa | Ser250 phosphorylation or Lys285 succinylation is possible; a visible shift is unproven. |
💡Expected NDUFA10 appearanceNDUFA10 has a predicted mass of 40.8 kDa and an observed band at ~41 kDa in reducing whole-cell blots; confirm band identity with ordinary controls.
| Predicted NDUFA10 mass | 40.8 kDa provides the sequence-based reference; the observed band is ~41 kDa. |
| Isoforms 1 and 2 | May differ in size, but their masses and migration are not supplied. |
| Ser250 phosphorylation by PINK1 | Could alter mobility; no visible size shift is established. |
| Lys285 succinylation | Could alter mobility; no visible size shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or no signal | NDUFA10 is localized to the mitochondrial matrix. | Check mitochondrial enrichment and sample loading. |
| Band higher than expected | A modification or unrelated band is possible; no upward shift is established. | Compare with the ~41 kDa reference and confirm identity with an independent antibody. |
| Band lower than expected | An isoform or degradation is possible; neither has a supplied band size. | Check sample integrity and confirm identity with an independent antibody. |
| Multiple bands | Isoforms 1 and 2 or unrelated antibody binding may contribute. | Compare mitochondrial enrichment and use an independent antibody. |
| Fragments below expected size | Protein degradation is possible; no fragment mass is supplied. | Review sample handling and compare freshly prepared lysate. |
Experiment evidence
Western blot images showing NDUFA10 band
A10517-1 · NDUFA10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NDUFA10 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NDUFA10 in Western blot, you can use adrenal gland lysate, rated Medium by HPA.
Positive control: Adrenal gland (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: HPA rates the positive tissues only Medium, so a strong positive signal may be difficult to obtain.
HPA tissue expression evidence for NDUFA10
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | 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 → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NDUFA10 Western Blot Tips
Deeper troubleshooting and optimisation questions for NDUFA10, answered from its protein features.
How should NDUFA10 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 NDUFA10 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 has a replacement at residue 183, lacks residues 214–223, and has a different sequence at residues 334–355. These changes could affect apparent size or antibody recognition, but the features do not establish where either isoform runs.
How should Ser250 phosphorylation be assessed?
PTM · UniProt lists phosphoserine at position 250 by PINK1. Use that UniProt coordinate when selecting a site specific antibody, and verify its numbering against the antibody documentation. The feature does not establish that phosphorylation produces a resolvable band shift.
UniProt lists N6-succinyllysine at position 285. Check whether an antibody targets or spans that region when interpreting changes in signal. Position 285 uses the supplied UniProt numbering; the feature does not demonstrate a separate band.
UniProt lists N6-succinyllysine at position 285. Check whether an antibody targets or spans that region when interpreting changes in signal. Position 285 uses the supplied UniProt numbering; the feature does not demonstrate a separate band.
Does this guide establish induction of NDUFA10?
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.
How should transfer be checked for NDUFA10?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A10517-1 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 NDUFA10 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 observed NDUFA10 band near 41 kDa?
Interpretation · The observed band near 41 kDa agrees closely with the 40.8 kDa predicted mass. The listed modifications alone do not establish a visible shift or explain any small difference between predicted and apparent mass.
For total NDUFA10 quantitation, choose an antibody whose recognized sequence is shared by both listed isoforms, then compare signals within the assay's linear range. Isoform 2 changes residues 183, 214–223, and 334–355, so an antibody targeting those regions may measure the isoforms differently.
Check whether an additional band is consistent with the listed isoform 2 sequence changes and whether the antibody recognizes both isoforms. Ser250 phosphorylation and Lys285 succinylation are also listed, but their presence alone cannot identify an unexpected band or prove a visible mass shift.
For total NDUFA10 quantitation, choose an antibody whose recognized sequence is shared by both listed isoforms, then compare signals within the assay's linear range. Isoform 2 changes residues 183, 214–223, and 334–355, so an antibody targeting those regions may measure the isoforms differently.
Check whether an additional band is consistent with the listed isoform 2 sequence changes and whether the antibody recognizes both isoforms. Ser250 phosphorylation and Lys285 succinylation are also listed, but their presence alone cannot identify an unexpected band or prove a visible mass shift.
Boster reagents
NDUFA10 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-NDUFA10 Antibody Picoband®
Cat # A10517-1
The catalog reports A10517-1, a rabbit polyclonal anti-NDUFA10 antibody with human and rat reactivity. Its Western blot image shows an approximately 41 kDa band in human RT4, HepG2, MCF-7, and PC-3 whole cell lysates. No rat blot is supplied.
Which to pick: A10517-1 is the only listed option. Its Western blot image uses 30 µg of human cell lysate per lane and 0.5 µg/mL primary antibody. Rat reactivity is listed, but the supplied image shows human samples only.
Source: BosterBio NDUFA10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95299.
- Human Protein Atlas. NDUFA10 tissue expression.
- PMC8311465 — target-verified WB comparison
- PMC9226000 — target-verified WB comparison
- PMC6511432 — target-verified WB comparison