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- Table of Contents
SKI / Ski oncogene · Western blot design guide
Design a Western Blot for SKI
Real validated SKI Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SKI 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
SKI Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~80 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Bone marrow (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SKI Western Blot Protocols
The A01062 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog A01062) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01062 · (A) 1 and (B) 2 μg/mL (catalog A01062) |
| 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 SKI Western Blot Band Size?
SKI has a predicted mass of 80 kDa; listed phosphorylation sites could affect migration, but no empirical band size or visible shift is established.
| Band near 80 kDa | consistent with the predicted size of SKI; confirm its identity with antibody controls |
| Stronger near-80 kDa band in nuclear fraction | consistent with SKI's nuclear location |
| Faint band in cytosolic fraction | consistent with SKI's nuclear location |
| Doublet near 80 kDa | phosphorylation could contribute, but distinct migrating forms are not established |
💡Expected SKI appearanceUniProt predicts an approximately 80 kDa SKI band, but no empirical band size is supplied; confirm any candidate band with ordinary band-identity controls.
| UniProt predicted mass | places the expected SKI band near 80 kDa |
| Phosphoserine 383 | could affect migration; no visible shift is established |
| Phosphoserine 432 | could affect migration; no visible shift is established |
| Phosphoserine 480 | could affect migration; no visible shift is established |
| Phosphoserine 720 | could affect migration; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear SKI may be poorly represented in the preparation | check nuclear fraction recovery and include a positive-control lysate |
| Band higher than expected | phosphorylation could affect migration, but the cause is unproven | compare phosphatase-treated and untreated samples and verify band identity |
| Band lower than expected | degradation or nonspecific binding is possible; no cleavage feature is listed | prepare fresh lysate and verify the band with an independent SKI antibody |
| Multiple bands | differential phosphorylation or nonspecific binding is possible | compare phosphatase-treated samples and verify bands with an independent SKI antibody |
| Weak or no signal | nuclear SKI may be underrepresented in the loaded sample | assess nuclear fraction recovery and use a positive-control lysate |
Experiment evidence
Western blot images showing SKI band
A01062 · SKI
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SKI Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SKI in Western blot, you can use bone marrow, which HPA scores High.
Positive control: Bone marrow (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: SKI is nuclear, so nuclear extracts may give a stronger signal than whole-tissue lysates.
HPA tissue expression evidence for SKI
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Testis | spermatogonia 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 → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SKI Western Blot Tips
Deeper troubleshooting and optimisation questions for SKI, answered from its protein features.
How should SKI 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 SKI isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. These features do not support assigning multiple bands to SKI isoforms; verify the identity of each band before making that assignment.
Which phosphorylation sites are listed for SKI?
PTM · UniProt lists phosphoserines at positions 383, 432, 480 and 720. Use these UniProt coordinates when selecting or reporting site-specific measurements; paper or antibody numbering may use a different convention.
SKI has four listed phosphoserines, but their presence alone does not demonstrate a visible band shift or explain a difference from 80 kDa. Confirm band identity and phosphorylation experimentally before assigning a shifted band to this modification.
SKI has four listed phosphoserines, but their presence alone does not demonstrate a visible band shift or explain a difference from 80 kDa. Confirm band identity and phosphorylation experimentally before assigning a shifted band to this modification.
Does this guide establish induction of SKI?
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.
How should transfer be checked for SKI?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01062 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 SKI be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Should SKI migrate at its predicted 80 kDa?
Interpretation · The predicted mass is 80 kDa, but no observed band position is supplied. Use 80 kDa as a starting point for identifying SKI; the listed features do not establish an apparent mass or a visible shift.
SKI is listed in the nucleus. Include nuclear material in the sample, and keep the sample fraction consistent when comparing lanes.
Compare SKI signal from the same type of sample fraction across lanes and state how the signal was normalized. Its nuclear localization makes changes in nuclear content or fraction choice relevant to interpretation.
Start with the predicted 80 kDa mass, then verify band identity. The record supplies no observed band position or alternative isoform sequence. Its phosphoserines and Ubl conjugation keyword identify possible modifications, but neither establishes the cause of an unexpected band.
SKI is listed in the nucleus. Include nuclear material in the sample, and keep the sample fraction consistent when comparing lanes.
Compare SKI signal from the same type of sample fraction across lanes and state how the signal was normalized. Its nuclear localization makes changes in nuclear content or fraction choice relevant to interpretation.
Start with the predicted 80 kDa mass, then verify band identity. The record supplies no observed band position or alternative isoform sequence. Its phosphoserines and Ubl conjugation keyword identify possible modifications, but neither establishes the cause of an unexpected band.
Boster reagents
SKI 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-Ski oncogene Ski Antibody
Cat # A01062
the supplier A01062 is an anti-SKI antibody listed for human reactivity. Its Western blot image shows HeLa cell lysate tested at 1 and 2 μg/mL. The supplied evidence does not establish performance in other specimens.
Which to pick: A01062 is the only listed option for SKI. Choose it when the human reactivity listing and the HeLa lysate Western blot example fit your experiment; the supplied evidence does not compare alternatives.
Source: BosterBio SKI gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P12755.
- Human Protein Atlas. SKI tissue expression.
- PMC6910224 — target-verified WB comparison
- PMC3919796 — target-verified WB comparison
- PMC11967118 — target-verified WB comparison