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- Table of Contents
DGAT2 / Diacylglycerol O-acyltransferase 2 · Western blot design guide
Design a Western Blot for DGAT2
Real validated DGAT2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-DGAT2 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
DGAT2 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 | ~43.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Membrane extraction controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated DGAT2 Western Blot Protocols
The A03174 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | C6 (40ug), AML-12 (40ug), A375 (40ug) (catalog A03174) |
| Gel % | 12–15% (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 | A03174; use the WB datasheet starting dilution (standard starting point) |
| 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 DGAT2 Western Blot Band Size?
DGAT2 is predicted at 43.8 kDa; isoforms and multimeric complexes could affect bands, but their migration has not been demonstrated here.
| Band near 43.8 kDa | Consistent with the predicted DGAT2 mass; confirm identity with antibody controls |
| Bands at different sizes | Could reflect isoforms 1 and 2, whose migration has not been established |
| Higher-mass band | Could reflect retained complexes containing several DGAT2 subunits |
| Weak band in a soluble lysate fraction | Could reflect DGAT2 association with endoplasmic reticulum membranes and lipid droplets |
💡Expected DGAT2 appearanceDGAT2 has a predicted mass of 43.8 kDa; no empirical band size is supplied, and any band assignment needs ordinary identity controls.
| UniProt predicted mass | Places the reference monomer near 43.8 kDa |
| Splice isoform 1 | May migrate differently from isoform 2; its mass is not supplied |
| Splice isoform 2 | May migrate differently from isoform 1; its mass is not supplied |
| Multimeric DGAT2 complexes | Could yield a higher apparent mass if complexes persist during electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated DGAT2 may be poorly recovered | Check membrane extraction and include a positive control lysate |
| Band higher than expected | DGAT2 complexes may persist, though their migration is unverified | Check denaturation and confirm identity with an independent antibody or DGAT2 depletion |
| Band lower than expected | An alternative isoform is possible, but its mass is unknown | Check isoform expression and confirm band identity with DGAT2 depletion |
| Multiple bands | Alternative isoforms or retained DGAT2 complexes are possible | Compare denaturing conditions and confirm bands with DGAT2 depletion |
| Weak or no signal | Membrane-associated DGAT2 may be underrepresented in the prepared fraction | Assess membrane protein recovery and use a positive control lysate |
| Fragments below expected size | DGAT2 may have degraded during sample preparation | Repeat with fresh lysate and protease inhibitors; verify fragment identity |
Experiment evidence
Western blot images showing DGAT2 band
A03174 · DGAT2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for DGAT2 Western blot
🧪For positive controls for DGAT2 in Western blot, you can use no HPA-supported tissue or cell sample because no HPA expression data were 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 alongside the samples.
⚠️Feasibility: HPA data do not identify a positive or negative tissue, so verify controls experimentally.
HPA tissue expression evidence for DGAT2
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 DGAT2 Western Blot Tips
Deeper troubleshooting and optimisation questions for DGAT2, answered from its protein features.
How should DGAT2 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 DGAT2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 41–83 in UniProt numbering, so its polypeptide is shorter. This feature does not establish whether the isoforms resolve as separate bands; check which isoform the antibody recognizes.
Can glycosylation explain an unexpected DGAT2 band?
PTM · The supplied UniProt features list no glycosylation sites or modified residues. They provide no basis to attribute an unexpected band to either modification; verify band identity and consider the two listed isoforms.
Does this guide establish induction of DGAT2?
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 DGAT2 Western blot?
Transfer · DGAT2 is a multi-pass endoplasmic reticulum membrane protein. Choose and check transfer conditions for membrane-protein recovery, then inspect the post-transfer gel and membrane to see whether DGAT2-sized material transferred. The supplied features do not establish a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03174 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 DGAT2 bands be quantified across samples?
Quantitation · Use comparable preparations that retain endoplasmic reticulum membrane and lipid-droplet material, and normalize against an appropriate loading measure for the fraction analyzed. If two bands appear, establish which represents DGAT2 or its isoforms before combining their intensities.
Should DGAT2 migrate at its predicted 43.8 kDa?
Interpretation · 43.8 kDa is the predicted mass, but no observed band position is supplied. DGAT2 is a multi-pass membrane protein, so confirm band identity independently rather than assigning it by apparent mass alone.
DGAT2 is found at the endoplasmic reticulum membrane and lipid droplets, with perinuclear cytoplasmic localization also reported. Check whether your preparation retains these compartments before interpreting a weak or absent band.
Isoform 2 lacks UniProt residues 41–83, making isoform expression one possibility. DGAT2 also forms multimeric complexes by similarity. Neither feature proves the identity of a particular band; compare isoform coverage and confirm specificity before assigning bands.
DGAT2 is found at the endoplasmic reticulum membrane and lipid droplets, with perinuclear cytoplasmic localization also reported. Check whether your preparation retains these compartments before interpreting a weak or absent band.
Isoform 2 lacks UniProt residues 41–83, making isoform expression one possibility. DGAT2 also forms multimeric complexes by similarity. Neither feature proves the identity of a particular band; compare isoform coverage and confirm specificity before assigning bands.
Boster reagents
DGAT2 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-DGAT2 Antibody
Cat # A03174
The catalog reports one anti-DGAT2 antibody, A03174, with stated Human, Mouse, and Rat reactivity. Its WB image shows C6, AML-12, A375, L02, and HepG2 whole-cell lysates at 40 µg per lane using a 1:500 antibody dilution. Broader sample validation is not shown.
Which to pick: A03174 is the only listed option and has a WB image. Check its stated reactivity and pictured samples and conditions against your experiment; the image documents those five lysates only.
Source: BosterBio DGAT2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96PD7.
- Human Protein Atlas. DGAT2 tissue expression.
- PMC8539991 — target-verified WB comparison
- PMC8953714 — target-verified WB comparison
- PMC12487672 — target-verified WB comparison