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- Table of Contents
SOAT1 / Sterol O-acyltransferase 1 · Western blot design guide
Design a Western Blot for SOAT1
Source-linked SOAT1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SOAT1 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
SOAT1 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 | ~64.7 kDa | |
| Observed band | ~75 kDa | |
| Gel | 5–20% (catalog A05096-1) | |
| Positive control | Adrenal gland (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked SOAT1 Western Blot Protocol Options
The A05096-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HUH-7, human THP-1, human HepG2, rat liver, mouse liver (catalog A05096-1) |
| Gel % | 5–20% (catalog A05096-1) |
| Load | 30 ug; reducing conditions (catalog A05096-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05096-1) |
| Membrane | nitrocellulose membrane (catalog A05096-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05096-1) |
| Primary antibody | A05096-1 · 0.5 μg/mL (catalog A05096-1) |
| Primary incubation | overnight at 4°C (catalog A05096-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05096-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05096-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05096-1) |
| Detection | ECL (catalog A05096-1) |
Section 2
What Is the Expected SOAT1 Western Blot Band Size?
SOAT1 is predicted at 64.7 kDa and observed near 75 kDa in reducing blots; the cause of the difference is not established.
| Band near 75 kDa | Empirical SOAT1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 65 kDa | Near the 64.7 kDa predicted mass; identity requires confirmation |
| Higher-mass band | Could reflect retained SOAT1 homooligomers if denaturation is incomplete |
| Multiple bands | SOAT1 has isoforms 1, 2 and 3, but distinct band positions are unestablished |
| Weak band in soluble lysate | SOAT1 is a multi-pass endoplasmic reticulum membrane protein that may be poorly recovered |
💡Expected SOAT1 appearanceSOAT1 has a predicted mass of 64.7 kDa, while reducing whole-cell blots show an empirical band near 75 kDa; its identity should be checked with ordinary controls, and the migration difference is unexplained.
| Predicted SOAT1 mass | 64.7 kDa is the sequence-based reference, not its demonstrated migration |
| Splice isoform 1 | Its distinct apparent size is not supplied |
| Splice isoform 2 | Its size relative to isoform 1 is not supplied |
| Splice isoform 3 | Its size relative to the other isoforms is not supplied |
| Homodimer formation | Retained assemblies could produce a higher-mass band if denaturation is incomplete |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Poor recovery of multi-pass SOAT1 from endoplasmic reticulum membranes | Check membrane solubilization and a membrane-enriched fraction |
| Band higher than expected | The reported 75 kDa band exceeds the 64.7 kDa prediction for an undetermined reason | Compare with a positive-control lysate and verify specificity by SOAT1 depletion |
| Band lower than expected | An unverified band may be a fragment or another protein | Check SOAT1 depletion and compare antibodies against different epitopes |
| Broad smear instead of sharp band | Incomplete solubilization of membrane-associated SOAT1 is possible | Optimize membrane extraction and sample denaturation |
| Multiple bands | Isoforms or incompletely denatured assemblies are possible, but their migrations are unestablished | Check reduction and denaturation, then verify bands by SOAT1 depletion |
| Weak or no signal | Membrane-associated SOAT1 may be underrepresented in the prepared sample | Check membrane protein recovery and use a positive-control lysate |
Experiment evidence
Western blot images showing SOAT1 band
A05096-1 · SOAT1
A05096 · SOAT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SOAT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SOAT1 in Western blot, you can use adrenal gland tissue, which HPA rates as highly expressed.
Positive control: Adrenal gland (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: As an ER multi-pass membrane protein, SOAT1 may require membrane-enriched lysate for a strong signal.
HPA tissue expression evidence for SOAT1
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 |
|---|---|---|---|---|
| Adrenal gland | cells in zona fasciculata | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | High | Protein (IHC) | HPA → |
| Skin | sebaceous glands | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SOAT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SOAT1, answered from its protein features.
How should SOAT1 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.
How could SOAT1 isoforms affect band interpretation?
Isoforms · Three isoforms are listed. Relative to canonical isoform 1, isoform 3 lacks residues 1–65; isoform 2 replaces residues 1–59 with methionine. These sequence differences could affect band positions and antibody recognition. Check whether the antibody targets the shared sequence before assigning a band to an isoform.
Which SOAT1 modifications matter when interpreting bands?
PTM · The listed UniProt modifications are N-acetylmethionine at position 1 and phosphoserine at position 8, using canonical sequence coordinates. Position 8 is absent from isoform 3 and lies within the sequence replaced in isoform 2. Neither modification alone establishes a visible shift. State the coordinate convention when comparing antibody or paper numbering.
Does this guide establish induction of SOAT1?
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 SOAT1 Western blot?
Transfer · SOAT1 is a 550-residue, multi-pass ER membrane protein with a predicted mass of 64.7 kDa. The supplied features do not specify a transfer method. Evaluate transfer of the approximately 75 kDa band by checking the membrane and post-transfer gel, then adjust transfer conditions if protein remains in the gel.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05096-1 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 SOAT1 bands be quantified across samples?
Quantitation · SOAT1 is an ER membrane protein, so use consistent membrane-protein extraction and sample preparation across samples. Quantify the same resolved band in each lane and confirm that the antibody detects the intended isoform or isoforms; three are listed. The supplied features do not identify which isoforms contribute to the observed approximately 75 kDa band.
Why does SOAT1 appear near 75 kDa instead of 64.7 kDa?
Interpretation · The reported band is about 75 kDa, while the predicted mass is 64.7 kDa. SOAT1 is a multi-pass ER membrane protein, but the supplied features do not establish why it migrates at 75 kDa. Do not attribute the difference to its listed modifications without further evidence.
SOAT1 forms a homotetramer composed of two homodimers and has one listed disulfide bond. Higher bands could warrant checking whether sample preparation preserves associated or disulfide-linked species. These features alone do not identify an unexpected band; compare reducing and nonreducing preparations before assigning it.
SOAT1 forms a homotetramer composed of two homodimers and has one listed disulfide bond. Higher bands could warrant checking whether sample preparation preserves associated or disulfide-linked species. These features alone do not identify an unexpected band; compare reducing and nonreducing preparations before assigning it.
Boster reagents
SOAT1 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-SOAT1 Antibody Picoband®
Cat # A05096-1
Real WB data
Anti-Sterol O-acyltransferase 1 SOAT1 Antibody
Cat # A05096
Both the supplier anti-SOAT1 antibodies list human, mouse, and rat reactivity and have WB images. A05096-1 documents human cell and rat and mouse liver lysates, with a band near 75 kDa versus 65 kDa expected. A05096 has only a brief WB description.
Which to pick: Choose A05096-1 if you want documented WB samples and conditions. A05096 lists the same species reactivity but provides no sample or protocol details in the supplied description.
Source: BosterBio SOAT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P35610.
- Human Protein Atlas. SOAT1 tissue expression.
- PMC11082151 — target-verified WB comparison
- PMC11234219 — target-verified WB comparison
- PMC7478739 — target-verified WB comparison