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- Table of Contents
ATOX1 / Copper transport protein ATOX1 · Western blot design guide
Design a Western Blot for ATOX1
Source-linked ATOX1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ATOX1 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
ATOX1 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 | ~7.4 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ATOX1 Western Blot Protocol Options
The A03421 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A03421) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A03421) |
| Primary antibody | A03421 · 1:1000 (catalog A03421) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A03421) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A03421) |
Section 2
What Is the Expected ATOX1 Western Blot Band Size?
ATOX1 is predicted at 7.4 kDa; homodimerization and documented modifications could affect patterns, but their effects on migration are not demonstrated here.
| Single band near 7.4 kDa | Consistent with the predicted ATOX1 monomer; confirm identity with controls |
| Band near twice 7.4 kDa | Could reflect a homodimer that survives sample preparation |
| Bands near 7.4 kDa and twice that size | Could reflect monomer and surviving homodimer |
| Slightly shifted band near 7.4 kDa | May reflect ATOX1 modification, but a visible shift is unproven |
💡Expected ATOX1 appearanceATOX1 has a predicted monomer mass of 7.4 kDa; no empirical band position is supplied, so confirm any candidate band with appropriate identity controls.
| Predicted monomer mass | Places the unmodified ATOX1 monomer near 7.4 kDa |
| Homodimer formation | Could yield a band near twice the monomer size if the dimer survives preparation |
| Phosphoserine 47 | May alter migration, but no visible size shift is established |
| N6-acetyllysine 60 | May alter migration, but no visible size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The small ATOX1 protein may be lost during transfer | Check transfer and membrane retention with a suitable control |
| Band higher than expected | ATOX1 homodimer may survive sample preparation | Compare denaturing conditions and verify band identity |
| Band lower than expected | A lower band is not explained by the supplied ATOX1 features | Verify identity with an independent antibody or ATOX1 depletion |
| Multiple bands | Monomer and surviving homodimer are possible, but identities are unconfirmed | Compare sample preparation conditions and use ATOX1 depletion |
| Weak or no signal | A 7.4 kDa protein may transfer through the membrane | Check membrane retention and optimize transfer for small proteins |
Experiment evidence
Western blot images showing ATOX1 band
A03421 · ATOX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ATOX1 Western blot
🧪For positive controls for ATOX1 in Western blot, you can use no HPA-supported tissue or cell line because expression data were not supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA data are unavailable, so tissue-based positive and negative controls cannot be selected.
HPA tissue expression evidence for ATOX1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced ATOX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for ATOX1, answered from its protein features.
How should ATOX1 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 ATOX1 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Which ATOX1 modifications should I consider when interpreting a band?
PTM · UniProt lists phosphoserine at position 47 and N6-acetyllysine at position 60; these are UniProt coordinates and may differ from antibody or paper numbering. Their presence does not show that either modification produces a resolvable Western-blot band shift.
Does this guide establish induction of ATOX1?
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 ATOX1 Western blot?
Transfer · ATOX1 is small, at a predicted 7.4 kDa. Choose a transfer setup suitable for retaining low-molecular-weight proteins and verify retention with an appropriate low-range marker. The supplied features do not specify a membrane or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03421 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 I quantify an ATOX1 Western-blot signal?
Quantitation · Quantify the band assigned to ATOX1 near its predicted 7.4 kDa mass, using consistent band boundaries and signals within the assay's linear range. The supplied features give no observed band position or condition-dependent abundance change, so confirm band identity before comparing samples.
Why might ATOX1 migrate differently from its predicted mass?
Interpretation · ATOX1 is 68 amino acids with a predicted mass of 7.4 kDa. UniProt lists phosphoserine at position 47 and N6-acetyllysine at position 60, using UniProt numbering. These features alone do not establish a visible shift or explain any apparent mass difference; no observed band size was supplied.
UniProt describes ATOX1 as a homodimer. A band near twice the predicted monomer mass, about 14.8 kDa, could prompt a dimer check, but the homodimer annotation does not establish that it survives Western-blot sample preparation or explains an observed band.
UniProt describes ATOX1 as a homodimer. A band near twice the predicted monomer mass, about 14.8 kDa, could prompt a dimer check, but the homodimer annotation does not establish that it survives Western-blot sample preparation or explains an observed band.
Boster reagents
ATOX1 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-Copper transport protein ATOX1 ATOX1 Antibody
Cat # A03421
The catalog reports one anti-ATOX1 antibody for Western blotting: A03421, with reported human reactivity and a WB image. Its caption describes cell-line extracts tested at 1:1000 dilution, with 25 µg protein per lane; it does not identify the cell lines.
Which to pick: A03421 is the only listed option. Its WB image and reported human reactivity make it the relevant choice for human samples; the supplied caption does not establish performance in other species.
Source: BosterBio ATOX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00244.
- Human Protein Atlas. ATOX1 tissue expression.
- PMC12265579 — target-verified WB comparison
- PMC13527614 — target-verified WB comparison
- PMC12375821 — target-verified WB comparison