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- Table of Contents
COX8A / Cytochrome c oxidase subunit 8A, mitochondrial · Western blot design guide
Design a Western Blot for COX8A
Source-linked COX8A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-COX8A 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
COX8A 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 | ~7.6 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Colon (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 | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked COX8A Western Blot Protocol Options
The A12782 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A549, sp2/0, Rat brain (catalog A12782) |
| Gel % | 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 | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A12782; use the WB datasheet starting dilution (standard starting point) |
| 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 COX8A Western Blot Band Size?
COX8A is predicted at 7.6 kDa; no empirical band is supplied, and its annotated features do not demonstrate altered migration.
| Band near 7.6 kDa | consistent with the predicted COX8A mass; confirm identity with antibody controls |
| Little or no band in whole-cell lysate | COX8A may be poorly recovered from the mitochondrial inner membrane |
| Stronger band in a mitochondrial membrane fraction | consistent with COX8A localization |
| Additional band away from 7.6 kDa | identity is uncertain without further controls |
💡Expected COX8A appearanceUniProt predicts COX8A at 7.6 kDa; no empirical band size is supplied, so confirm a band near that size with antibody specificity controls.
| UniProt predicted mass | places the expected band near 7.6 kDa |
| Predicted molecular weight of 7579 Da | corresponds to approximately 7.6 kDa |
| Predicted COX8A mass | provides a reference for assessing band position |
| Predicted protein size | does not establish actual gel migration |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | COX8A resides in the mitochondrial inner membrane | check membrane recovery and compare with an enriched fraction |
| Weak or no signal | recovery of this small membrane protein may be limited | check extraction, transfer and antibody performance |
| Band higher than expected | the supplied features do not establish a cause | check denaturation and confirm identity with an independent antibody or knockdown control |
| Band lower than expected | the supplied features do not establish a smaller product | check for degradation and confirm identity with an independent antibody or knockdown control |
| Multiple bands | the single annotated isoform does not establish multiple COX8A bands | compare bands with an independent antibody or knockdown control |
Experiment evidence
Western blot images showing COX8A band
A12782 · COX8A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for COX8A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for COX8A in Western blot, you can use colon tissue lysate.
Positive control: Colon (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.
⚠️Feasibility: COX8A is an inner mitochondrial membrane protein, so a mitochondria-enriched lysate may improve detection.
HPA tissue expression evidence for COX8A
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 |
|---|---|---|---|---|
| Colon | peripheral nerve/ganglion | High | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
| Skin | keratinocytes | High | Protein (IHC) | HPA → |
| Tonsil | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Appendix | glandular 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced COX8A Western Blot Tips
Deeper troubleshooting and optimisation questions for COX8A, answered from its protein features.
How should COX8A 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 COX8A isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands.
Are COX8A modifications expected to shift its Western blot band?
PTM · No modified residues or glycosylation sites are mapped in the supplied features. A Ubl conjugation keyword appears, but it gives no site or demonstrated band shift. Do not assign a shifted band to a specific modification from this record.
Does this guide establish induction of COX8A?
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 COX8A Western blot?
Transfer · COX8A is small (predicted 7.6 kDa) and a single-pass inner-membrane protein. Choose and check a transfer setup that retains small proteins; verify transfer and retention experimentally. The supplied features do not establish a specific membrane, transfer time, or voltage.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A12782 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 COX8A bands be quantified?
Quantitation · Quantify a band only after confirming it is specific and within the assay’s linear range. COX8A is a component of mitochondrial complex IV, so account for mitochondrial content when comparing samples. The supplied features provide no observed band position to identify automatically.
Why might COX8A migrate differently from its predicted 7.6 kDa?
Interpretation · The 7.6 kDa value is predicted for the 69-residue sequence. UniProt lists a transit peptide keyword but supplies no cleavage coordinates, and no observed band is available. Do not assign a specific mature mass or explain a band shift from these features alone.
Compare unexpected bands with the predicted 7.6 kDa sequence mass and check antibody specificity experimentally. COX8A belongs to a multisubunit complex and has a transit peptide keyword, but these features alone cannot identify another band or establish a processing-related shift.
Compare unexpected bands with the predicted 7.6 kDa sequence mass and check antibody specificity experimentally. COX8A belongs to a multisubunit complex and has a transit peptide keyword, but these features alone cannot identify another band or establish a processing-related shift.
Boster reagents
COX8A 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-COX8A Antibody
Cat # A12782
The catalog reports one anti-COX8A antibody for Western blotting, A12782. Its WB image shows A549 and sp2/0 whole-cell lysates and rat brain tissue lysate at 1:500 dilution. The supplied evidence does not establish broader validation.
Which to pick: A12782 is the only listed option. It reports human, mouse, and rat reactivity and has a WB image with A549, sp2/0, and rat brain samples. Choose it if those reported contexts fit your experiment; other samples need validation.
Source: BosterBio COX8A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P10176.
- Human Protein Atlas. COX8A tissue expression.
- PMC9877034 — target-verified WB comparison
- PMC13343725 — target-verified WB comparison