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- Table of Contents
SETX / Helicase senataxin · Western blot design guide
Design a Western Blot for SETX
Source-linked SETX Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SETX 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
SETX 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 | ~302.9 kDa | |
| Observed band | ~303 kDa | |
| Gel | 8% (catalog A02233-1) | |
| 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 | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SETX Western Blot Protocol Options
The A02233-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human MCF-7 (catalog A02233-1) |
| Gel % | 8% (catalog A02233-1) |
| Load | 30 ug; reducing conditions (catalog A02233-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02233-1) |
| Membrane | nitrocellulose membrane (catalog A02233-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02233-1) |
| Primary antibody | A02233-1 · 0.5 μg/mL (catalog A02233-1) |
| Primary incubation | overnight at 4°C (catalog A02233-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02233-1) |
| Secondary incubation | 1.5 hour at RT (catalog A02233-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02233-1) |
| Detection | ECL (catalog A02233-1) |
Section 2
What Is the Expected SETX Western Blot Band Size?
SETX is predicted at 302.9 kDa and observed near 303 kDa; the small difference has no established cause in the supplied evidence.
| Band near 303 kDa | matches the empirical SETX band and its 302.9 kDa predicted mass |
| Higher band near twice the monomer size | could reflect a retained SETX homodimer if supported by band-identity controls |
| Additional bands near the main band | could reflect isoforms 1, 3, or 4 if their migration differs |
| Weak band in a cytoplasmic fraction | SETX also localizes to the nucleus and chromosomes |
💡Expected SETX appearanceSETX has a predicted mass of 302.9 kDa and an observed band near 303 kDa in reducing whole-cell blots; confirm band identity with appropriate controls.
| Predicted SETX mass | 302.9 kDa, consistent with the observed band near 303 kDa |
| SETX homodimer | could yield a higher band if the dimer remains intact during electrophoresis |
| Isoforms 1, 3, and 4 | may differ in size, but their masses and migration differences are not supplied |
| Phosphoserine sites including Ser615 and Ser642 | may alter apparent migration; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SETX may be poorly recovered from nuclear or chromosome-associated material | check extraction and compare with a HeLa whole-cell positive control |
| Band higher than expected | a SETX homodimer may remain intact | check reduction and denaturation, then confirm band identity |
| Band lower than expected | a shorter isoform or protein breakdown is possible | compare antibody epitopes and use an independent SETX antibody |
| Multiple bands | isoforms 1, 3, and 4 or nonspecific binding are possible | compare isoform expression and confirm bands with an independent antibody |
| Weak or no signal | limited recovery of nuclear SETX is possible | check nuclear extraction, loading, and a HeLa positive control |
| Fragments below expected size | SETX may have broken down during sample preparation | prepare fresh lysate with protease inhibitors and confirm fragments with a second antibody |
Experiment evidence
Western blot images showing SETX band
A02233-1 · SETX
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SETX Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SETX in Western blot, you can use appendix tissue, which HPA scores as Medium.
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: SETX is nuclear, so nuclear enrichment may improve detection in tissue lysate.
HPA tissue expression evidence for SETX
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 | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Hippocampus | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SETX Western Blot Tips
Deeper troubleshooting and optimisation questions for SETX, answered from its protein features.
How should SETX 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 SETX isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 3, and 4. Isoform 3 lacks residues 2367–2399, while isoform 4 replaces M at position 2429 with MQLLPRSFCVHVNHSPFFSPEPKYLHWALK. Check whether the antibody epitope overlaps these regions. The sequence differences alone do not establish that separate bands will be visible.
Which SETX phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at 615, 642, 878, 911, 947, 956, 1017, 1019, 1330, 1366, 1489, 1621, 1623, and 1663, plus phosphothreonine at 2474. These are UniProt coordinates; antibody or paper numbering may differ. Site annotations alone cannot attribute a visible band shift to phosphorylation.
Does this guide establish induction of SETX?
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 SETX Western blot?
Transfer · SETX is approximately 303 kDa. Use a transfer setup validated for proteins of that size, and check the post-transfer gel and membrane for recovery before interpreting a weak signal. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02233-1 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 SETX bands be quantified across samples?
Quantitation · Quantify the same band and cellular fraction across samples. SETX is annotated in the nucleus and cytoplasm, so changes in fraction recovery could affect the measured signal. If multiple bands appear, establish which represent SETX before combining their intensities.
Why does SETX appear near 303 kDa?
Interpretation · SETX has a predicted mass of 302.9 kDa, consistent with the observed band near 303 kDa. Its listed features do not establish a modification-related shift.
Compare them with the approximately 303 kDa band and consider the listed isoform sequence differences and phosphorylation sites as possibilities to investigate. Confirm band identity with an independent SETX antibody or another specificity check; the supplied features alone cannot identify an unexpected band.
Compare them with the approximately 303 kDa band and consider the listed isoform sequence differences and phosphorylation sites as possibilities to investigate. Confirm band identity with an independent SETX antibody or another specificity check; the supplied features alone cannot identify an unexpected band.
Boster reagents
SETX 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-SETX Antibody Picoband®
Cat # A02233-1
A02233-1 is a rabbit polyclonal anti-SETX antibody listed for human samples. Its Western blot image shows an approximately 303 kDa band in HeLa and MCF-7 whole cell lysates. No independent validation evidence is supplied.
Which to pick: A02233-1 is the only listed option. Its Western blot image documents testing in human HeLa and MCF-7 lysates at 0.5 μg/mL; suitability for other species or sample types is not established by the supplied evidence.
Source: BosterBio SETX gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q7Z333.
- Human Protein Atlas. SETX tissue expression.
- PMC10580588 — target-verified WB comparison
- PMC11463457 — target-verified WB comparison
- PMC9536298 — target-verified WB comparison