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- Table of Contents
SCO2 / Cytochrome c oxidase assembly factor SCO2 · Western blot design guide
Design a Western Blot for SCO2
Source-linked SCO2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SCO2 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
SCO2 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 | ~29.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SCO2 Western Blot Protocol Options
The A02749 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HL60 cell lysate lysate (catalog A02749) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02749 · (A) 1 and (B) 2 μg/mL (catalog A02749) |
| 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 SCO2 Western Blot Band Size?
SCO2 has a predicted monomer mass of 29.8 kDa; homodimer formation may affect migration, but no empirical band size or demonstrated shift is supplied.
| Band near 29.8 kDa | Consistent with the predicted SCO2 monomer; confirm identity with antibody controls. |
| Band near 60 kDa | Could reflect a homodimer if the complex survives sample preparation. |
| Faint or absent band in whole-cell lysate | SCO2 is an inner mitochondrial membrane protein and may be poorly recovered. |
| Stronger band after mitochondrial membrane enrichment | Consistent with SCO2 localization to the mitochondrial inner membrane. |
💡Expected SCO2 appearanceUniProt predicts a 29.8 kDa SCO2 monomer, but no empirical band size is supplied; confirm any band with antibody and sample controls, especially if a homodimer persists.
| Predicted monomer mass | Places the sequence-based reference near 29.8 kDa. |
| Homodimer formation | Could produce a band near twice the monomer mass if the dimer persists during electrophoresis. |
| Intrachain disulfide between residues 133 and 137 | May affect migration under different reduction conditions; no size shift is established. |
| Mitochondrial inner membrane localization | Can affect protein recovery, but does not establish a different molecular size. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Inner mitochondrial membrane SCO2 may be poorly recovered. | Check membrane extraction and test a mitochondrial membrane-enriched fraction. |
| Band higher than expected | A SCO2 homodimer may persist during sample preparation. | Compare reducing and non-reducing preparations and verify band identity. |
| Band lower than expected | The band identity or protein integrity is uncertain. | Check antibody specificity and compare with a fresh sample. |
| Multiple bands | Monomer and persistent homodimer are possible, but identity is unconfirmed. | Compare reduction conditions and use an independent SCO2 antibody. |
| Weak or no signal | Recovery or detection of inner membrane SCO2 may be limited. | Check membrane enrichment, protein loading, and antibody performance. |
| Fragments below expected size | Protein degradation is possible; no fragment sizes are supplied. | Prepare fresh lysate with protease inhibitors and verify band identity. |
Experiment evidence
Western blot images showing SCO2 band
A02749 · SCO2
A02749-1 · SCO2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SCO2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SCO2 in Western blot, you can use adrenal gland tissue, which HPA scores High.
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 such as Ponceau alongside the samples.
⚠️Feasibility: SCO2 is an inner-membrane protein, so mitochondrial-enriched lysate may improve detection.
HPA tissue expression evidence for SCO2
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 | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SCO2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SCO2, answered from its protein features.
How should SCO2 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 multiple SCO2 bands represent isoforms?
Isoforms · Only one isoform is listed, with no alternative sequence. Do not assign additional bands to SCO2 isoforms based on this record.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of SCO2?
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 SCO2 Western blot?
Transfer · SCO2 is a 29.8 kDa, single-pass inner mitochondrial membrane protein. The supplied features do not specify a transfer method. Check transfer efficiency for the band of interest when optimizing the blot.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02749 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 SCO2 bands be quantified across samples?
Quantitation · SCO2 resides in the mitochondrial inner membrane. Use comparable membrane preparations and solubilization conditions across samples, and quantify the same band species consistently. Its listed homodimer and protein interactions make band identity important when comparing signals.
Does SCO2's predicted mass explain a shifted Western blot band?
Interpretation · The predicted mass is 29.8 kDa, but no observed band size is supplied. SCO2 is a single-pass inner mitochondrial membrane protein, and its features include a transit peptide keyword without cleavage coordinates. These features alone cannot establish a band shift or its cause.
One disulfide bond is listed, but its residues are not supplied. Compare reducing and nonreducing samples if band patterns differ; a disulfide bond alone does not predict a visible shift.
SCO2 is listed as a homodimer and associates with COA6 and a larger assembly complex. A higher band could be consistent with associated proteins, but size alone cannot identify it. Compare sample preparation conditions before assigning the band to SCO2.
One disulfide bond is listed, but its residues are not supplied. Compare reducing and nonreducing samples if band patterns differ; a disulfide bond alone does not predict a visible shift.
SCO2 is listed as a homodimer and associates with COA6 and a larger assembly complex. A higher band could be consistent with associated proteins, but size alone cannot identify it. Compare sample preparation conditions before assigning the band to SCO2.
Boster reagents
SCO2 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-SCO2 Antibody
Cat # A02749
Real WB data
Anti-SCO2 Antibody Picoband®
Cat # A02749-1
Two human-reactive anti-SCO2 antibodies have WB images: A02749 in HL60 lysate at 1 and 2 μg/mL, and A02749-1 in 293T, HepG2, HeLa, and PC-3 lysates at 0.5 μg/mL. Evidence is limited to these reported WB contexts.
Which to pick: Choose A02749 for an HL60 WB example, or A02749-1 for its four human cell lysate examples and detailed conditions. Both have WB images; the captions do not establish performance beyond the reported samples.
Source: BosterBio SCO2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43819.
- Human Protein Atlas. SCO2 tissue expression.
- PMC3504012 — target-verified WB comparison
- PMC3445839 — target-verified WB comparison
- PMC7726716 — target-verified WB comparison