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- Table of Contents
SQLE / Squalene monooxygenase · Western blot design guide
Design a Western Blot for SQLE
Real validated SQLE Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SQLE 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
SQLE 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 | ~63.9 kDa | |
| Observed band | ~64 kDa | |
| Gel | 10% (catalog A06763-1) | |
| Positive control | Soft tissue (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
Real Curated SQLE Western Blot Protocols
The A06763-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human HepG2, human 293T (catalog A06763-1) |
| Gel % | 10% (catalog A06763-1) |
| Load | 30 ug; reducing conditions (catalog A06763-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06763-1) |
| Membrane | nitrocellulose membrane (catalog A06763-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06763-1) |
| Primary antibody | A06763-1 · 1:1000 (catalog A06763-1) |
| Primary incubation | overnight at 4°C (catalog A06763-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06763-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06763-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06763-1) |
| Detection | ECL (catalog A06763-1) |
Section 2
What Is the Expected SQLE Western Blot Band Size?
SQLE is predicted at 63.9 kDa and observed at approximately 64 kDa; the cause of their small difference is not established.
| Band near 64 kDa | Matches the observed SQLE band and its 63.9 kDa predicted mass |
| Single sharp band near 64 kDa | Consistent with the reported SQLE band |
| Band near 64 kDa in a microsomal fraction | Consistent with SQLE localization to microsomal membranes |
| Little or no band after membrane depletion | Consistent with loss of membrane-associated SQLE |
💡Expected SQLE appearanceSQLE has a predicted mass of 63.9 kDa and an observed band at approximately 64 kDa; confirm its identity with an appropriate control.
| UniProt predicted mass of 63.9 kDa | Places full-length SQLE near 64 kDa |
| Full-length SQLE predicted mass | Provides the reference size for comparing bands |
| Predicted SQLE monomer mass | Is consistent with the observed band near 64 kDa |
| UniProt mass estimate for SQLE | Does not establish a separate higher- or lower-mass species |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated SQLE may be lost during sample preparation | Check membrane recovery and use a positive lysate control |
| Band higher than expected | The supplied features do not establish a higher-mass SQLE species | Compare with the 64 kDa band and confirm identity by SQLE knockdown |
| Band lower than expected | The supplied features do not establish a smaller SQLE species | Check sample integrity and confirm identity by SQLE knockdown |
| Multiple bands | Additional bands are not explained by the listed SQLE features | Use SQLE knockdown to identify the specific band |
| Weak or no signal | Membrane-associated SQLE may be poorly recovered | Check membrane recovery and include a positive lysate control |
Experiment evidence
Western blot images showing SQLE band
A06763-1 · SQLE
A06763 · SQLE
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SQLE Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SQLE in Western blot, you can use soft tissue, which HPA rates highly positive.
Positive control: Soft tissue (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 the samples.
⚠️Feasibility: SQLE is membrane-associated, so a membrane-enriched lysate may improve detection.
HPA tissue expression evidence for SQLE
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 |
|---|---|---|---|---|
| Soft tissue | fibroblasts | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Colon | endothelial 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 → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SQLE Western Blot Tips
Deeper troubleshooting and optimisation questions for SQLE, answered from its protein features.
How should SQLE 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 SQLE isoforms explain additional bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Do not assign additional bands to SQLE isoforms from this evidence alone; check their specificity experimentally.
Could SQLE modification cause an unexpected band shift?
PTM · UniProt lists ubiquitin-like conjugation as a keyword, but supplies no modified residue or glycosylation site. That annotation alone cannot establish a visible shift or identify a shifted band. Validate any additional band before assigning it to modified SQLE.
Does this guide establish induction of SQLE?
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 SQLE Western blot?
Transfer · SQLE is associated with ER and microsome membranes. Verify that the approximately 64 kDa protein transfers efficiently by checking both the membrane and remaining gel, then adjust transfer conditions if needed. The supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06763-1 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 SQLE abundance be quantified across samples?
Quantitation · Quantify an unsaturated signal around 64 kDa within the assay’s linear range. Compare samples prepared with consistent recovery of membrane-associated protein, since SQLE is located at ER and microsome membranes.
Should SQLE migrate at its predicted molecular mass?
Interpretation · SQLE has a predicted mass of 63.9 kDa, close to the supplied observed band of approximately 64 kDa. Use that region as a starting point for identification; the agreement alone does not establish band identity.
The supplied record lists one isoform, no glycosylation sites, and no modified residues. These features do not identify the source of other bands. Check band specificity, for example through SQLE depletion or a second antibody, before interpreting them as SQLE.
The supplied record lists one isoform, no glycosylation sites, and no modified residues. These features do not identify the source of other bands. Check band specificity, for example through SQLE depletion or a second antibody, before interpreting them as SQLE.
Boster reagents
SQLE 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-SQLE Antibody
Cat # A06763-1
Real WB data
Anti-Squalene monooxygenase SQLE Antibody
Cat # A06763
Both listed anti-SQLE antibodies have Western blot images and report human, mouse, and rat reactivity. The supplied blots use human cell lysates; no mouse or rat blot examples are shown. A06763-1 reports a band near the expected 64 kDa size.
Which to pick: Choose A06763-1 for a blot example across A549, HepG2, 293T, and MCF-7 lysates at 1:1000. A06763 has a HepG2 example at 1:1940. Both show HepG2; neither supplied image demonstrates mouse or rat samples.
Source: BosterBio SQLE gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14534.
- Human Protein Atlas. SQLE tissue expression.
- PMC12657232 — target-verified WB comparison
- PMC10404895 — target-verified WB comparison
- PMC9254471 — target-verified WB comparison