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- Table of Contents
STIL / SCL-interrupting locus protein · Western blot design guide
Design a Western Blot for STIL
Source-linked STIL Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-STIL 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
STIL 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 | ~143 kDa | |
| Observed band | ~170 kDa | |
| Gel | 5–20% (catalog A04956-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked STIL Western Blot Protocol Options
The A04956-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat testis, rat C6, rat PC-12, mouse NIH/3T3, human Hela (catalog A04956-1) |
| Gel % | 5–20% (catalog A04956-1) |
| Load | 30ug; reducing conditions (catalog A04956-1) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog A04956-1) |
| Membrane | Nitrocellulose membrane (catalog A04956-1) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog A04956-1) |
| Primary antibody | A04956-1 · 0.5 μg/mL (catalog A04956-1) |
| Primary incubation | overnight at 4°C (catalog A04956-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04956-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04956-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04956-1) |
| Detection | ECL (catalog A04956-1) |
Section 2
What Is the Expected STIL Western Blot Band Size?
STIL is predicted at 143 kDa, while a reducing Western blot reports approximately 170 kDa; the cause of the difference is not established.
| Band near 170 kDa | Empirical STIL band under reducing conditions; confirm identity with antibody controls |
| Band near 143 kDa | Near the predicted mass, but its identity requires confirmation |
| Band near twice the monomer size | Could represent the STIL homodimer if reduction is incomplete |
| Several bands at different positions | Isoforms 1 and 2 could contribute, but distinct migration is unproven |
💡Expected STIL appearanceSTIL has a predicted mass of 143 kDa, while an antibody QC blot reports a band near 170 kDa under reducing conditions; the cause of the difference is unestablished, so confirm identity with antibody controls.
| Predicted STIL mass | 143 kDa is the sequence-based reference, not the measured band position |
| STIL homodimer | Could yield a band near twice the monomer size if retained during electrophoresis |
| Isoform 1 | Its apparent size relative to isoform 2 is not supplied |
| Isoform 2 | Its apparent size relative to isoform 1 is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The reported STIL band is near 170 kDa despite a 143 kDa predicted mass; the cause is unestablished | Compare with the QC blot and confirm identity using antibody controls |
| Band lower than expected | A lower band could be a different isoform, degradation product, or nonspecific signal | Check sample integrity and confirm band identity with antibody controls |
| Multiple bands | STIL has isoforms 1 and 2, though their migration is not established | Confirm each band with antibody controls |
| Weak or no signal | STIL may be difficult to detect in the tested lysate | Check lysate quality, loading, and a positive control |
| No band in lysate | The tested lysate may lack detectable STIL | Use a positive control and check antibody performance |
Experiment evidence
Western blot images showing STIL band
A04956-1 · STIL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for STIL Western blot
🧪For positive controls for STIL in Western blot, you can use a verified STIL-positive cell lysate, although no HPA positive tissue or cell line is 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 such as Ponceau alongside the samples.
⚠️Feasibility: HPA supplies no tissue data, so tissue-based controls cannot be selected from this evidence.
HPA tissue expression evidence for STIL
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 STIL Western Blot Tips
Deeper troubleshooting and optimisation questions for STIL, answered from its protein features.
How should STIL 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 STIL isoforms explain an unexpected band?
Isoforms · UniProt lists isoforms 1 and 2. At canonical position 872, isoform 2 has NS in place of N. This one-residue insertion is unlikely to account for the 143 versus approximately 170 kDa difference. Check whether the antibody recognizes both isoforms before assigning separate bands.
Which STIL modifications matter when interpreting band mobility?
PTM · UniProt annotates N-acetylmethionine at position 1 and phosphoserines at positions 395, 753, 779, and 1135. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish a visible mobility shift. Compare phosphorylation-sensitive and control samples if testing a shift.
Does this guide establish induction of STIL?
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 STIL Western blot?
Transfer · Check transfer efficiency around the observed approximately 170 kDa band, using a suitable molecular-weight marker or total-protein check. The supplied features do not identify a specific optimal transfer method. Verify transfer before interpreting a weak or missing high-mass band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04956-1 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 STIL Western blot signals be quantified?
Quantitation · Quantify the same validated STIL band across matched samples, using a consistent loading reference. STIL is annotated in the cytosol and at centrioles, centrosomes, and the cell cortex, so keep the sampled fraction consistent. Do not combine distinct bands as STIL without confirming their identity.
Why does STIL appear near 170 kDa instead of 143 kDa?
Interpretation · The supplied predicted mass is 143 kDa, while the observed band is about 170 kDa. STIL has annotated phosphorylation and acetylation sites, but those features alone do not explain the difference. Use the observed position as a reference and confirm band identity independently.
The supplied interaction annotation says PIN1 binding depends on STIL mitotic phosphorylation. Record and match cell-cycle conditions when comparing bands. This annotation supports a phosphorylation-dependent interaction, but does not establish a change in STIL abundance or band position.
Do not assign smaller bands to signal-peptide or propeptide cleavage: neither feature is annotated. The listed isoform change is only a one-residue insertion at canonical position 872. Confirm band identity before attributing smaller bands to STIL processing, an isoform, or degradation.
The supplied interaction annotation says PIN1 binding depends on STIL mitotic phosphorylation. Record and match cell-cycle conditions when comparing bands. This annotation supports a phosphorylation-dependent interaction, but does not establish a change in STIL abundance or band position.
Do not assign smaller bands to signal-peptide or propeptide cleavage: neither feature is annotated. The listed isoform change is only a one-residue insertion at canonical position 872. Confirm band identity before attributing smaller bands to STIL processing, an isoform, or degradation.
Boster reagents
STIL 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-STIL/SIL Antibody Picoband®
Cat # A04956-1
The catalog reports one anti-STIL/SIL antibody, A04956-1, with stated human, mouse, and rat reactivity. Its Western blot image shows an approximately 170 kDa band in rat testis, rat C6 and PC-12, mouse NIH/3T3, and human HeLa lysates.
Which to pick: A04956-1 is the only listed option and has a Western blot image. Its caption reports 30 µg lysate per lane and 0.5 µg/mL primary antibody; use the shown samples and conditions to judge its fit for your experiment.
Source: BosterBio STIL gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15468.
- Human Protein Atlas. STIL tissue expression.
- PMC13096552 — target-verified WB comparison
- PMC8416176 — target-verified WB comparison
- PMC5830536 — target-verified WB comparison