This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
NDUFS2 / NADH dehydrogenase [ubiquinone] iron-sulfur protein 2, mitochondrial · Western blot design guide
Design a Western Blot for NDUFS2
Real validated NDUFS2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NDUFS2 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
NDUFS2 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 | ~52.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Acetylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated NDUFS2 Western Blot Protocols
The A05618-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysates from A431, Hela cell line, mouse brain and rat brain tissue lysate (from left to right), (catalog A05618-1) |
| 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 | A05618-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-rabbit IgG, 1:10000 (catalog A05618-1) |
| 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 NDUFS2 Western Blot Band Size?
NDUFS2 has a predicted mass of 52.5 kDa; annotated isoforms could affect migration, but no distinct bands or empirical migration are demonstrated.
| Band near 52.5 kDa | consistent with the predicted NDUFS2 mass; confirm band identity |
| Several bands at different positions | isoforms 1 and 2 are possible contributors; their migration is unknown |
| Band away from 52.5 kDa | an alternative isoform is possible, but its size is not supplied |
| Little or no band in whole-cell lysate | NDUFS2 detection may depend on recovery of mitochondrial inner membrane protein |
💡Expected NDUFS2 appearanceUniProt predicts 52.5 kDa for NDUFS2; no empirical band size is supplied, and any band near that position requires ordinary identity controls.
| UniProt predicted mass | provides a 52.5 kDa reference, not a measured band position |
| Alternative splicing | could change apparent size, but no migration difference is supplied |
| Isoform 1 | its individual mass and migration are not supplied |
| Isoform 2 | its individual mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | mitochondrial inner membrane protein may be poorly recovered | check mitochondrial enrichment and lysate preparation |
| Band lower than expected | isoform identity or sample degradation is unresolved | compare samples and verify identity with a second antibody |
| Multiple bands | isoforms 1 and 2 are annotated, but their band positions are unknown | check specificity with NDUFS2 knockdown or knockout |
| Weak or no signal | insufficient recovery of mitochondrial protein is possible | check loading and mitochondrial recovery |
| Fragments below expected size | sample degradation is possible; fragment masses are not supplied | repeat with fresh, protease-protected lysate and verify specificity |
Experiment evidence
Western blot images showing NDUFS2 band
A05618-1 · NDUFS2
A05618-3 · NDUFS2
M05618-1 · NDUFS2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NDUFS2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NDUFS2 in Western blot, you can use appendix tissue, which HPA rates High.
Positive control: Appendix (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, NDUFS2 requires effective mitochondrial protein extraction.
HPA tissue expression evidence for NDUFS2
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NDUFS2 Western Blot Tips
Deeper troubleshooting and optimisation questions for NDUFS2, answered from its protein features.
How should NDUFS2 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 NDUFS2 isoforms produce separate bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, canonical residues 454–463 (QDIVFGEVDR) are replaced by RPIV, shortening the C-terminus by six residues. Separate bands are possible but are not established by this sequence difference alone; check whether the antibody epitope includes this region.
Do NDUFS2 modifications explain a shifted band?
PTM · UniProt lists N6-acetyllysine at position 62 and symmetric dimethylarginine at position 118, using canonical-sequence numbering. Their presence does not establish a visible band shift. Confirm a modification-specific assignment with an appropriate independent assay.
Does this guide establish induction of NDUFS2?
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 NDUFS2 Western blot?
Transfer · NDUFS2 is a 52.5 kDa peripheral protein on the matrix side of the mitochondrial inner membrane. The supplied features do not specify a transfer method. Check transfer near 52.5 kDa with a molecular-weight marker and a membrane stain, then adjust transfer conditions if signal remains in the gel.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05618-1 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 NDUFS2 bands be quantified?
Quantitation · NDUFS2 is a core Complex I subunit in the mitochondrial inner membrane. Compare like-for-like sample fractions and use a loading reference appropriate to the fraction. Quantify exposures within the detection system’s linear range.
Where should NDUFS2 migrate relative to its predicted mass?
Interpretation · The predicted mass is 52.5 kDa; no observed band position is supplied. Compare the signal with a molecular-weight marker, but do not assign any difference between apparent and predicted mass to a specific feature without further evidence.
Check whether an additional band could reflect isoform 2’s C-terminal replacement at canonical positions 454–463, and whether the antibody recognizes that region. The listed acetylation and methylation sites alone do not establish shifted bands. No observed band positions are supplied, so band identity requires experimental confirmation.
Check whether an additional band could reflect isoform 2’s C-terminal replacement at canonical positions 454–463, and whether the antibody recognizes that region. The listed acetylation and methylation sites alone do not establish shifted bands. No observed band positions are supplied, so band identity requires experimental confirmation.
Boster reagents
NDUFS2 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-NDUFS2 Antibody (Center)
Cat # A05618-1
Real WB data
Anti-NDUFS2 Antibody Picoband®
Cat # A05618-3
Real WB data
Anti-NDUFS2 Rabbit Monoclonal Antibody
Cat # M05618-1
All three listed anti-NDUFS2 antibodies have Western blot images. A05618-1 was tested on human cell lines and mouse and rat brain lysates; A05618-3 and M05618-1 were tested on named human, mouse, and rat samples. The latter two report bands below the expected 53 kDa.
Which to pick: Match the antibody to your sample: A05618-1 includes brain lysates; A05618-3 includes brain and lung lysates across the named species; M05618-1 includes lung lysates and additional cell lines. All three have WB images. Check the reported band position when choosing A05618-3 or M05618-1.
Source: BosterBio NDUFS2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75306.
- Human Protein Atlas. NDUFS2 tissue expression.
- PMC11715544 — target-verified WB comparison
- PMC9455994 — target-verified WB comparison
- PMC13118776 — target-verified WB comparison