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- Table of Contents
VGLL2 / Transcription cofactor vestigial-like protein 2 · Western blot design guide
Design a Western Blot for VGLL2
Source-linked VGLL2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VGLL2 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
VGLL2 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 | ~33.4 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 | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked VGLL2 Western Blot Protocol Options
The A11724 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | The Heart of Rat(40ug), The Heart of Mouse(40ug), Hela (40ug) (catalog A11724) |
| 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 | A11724; 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 VGLL2 Western Blot Band Size?
VGLL2 is predicted at 33.4 kDa; its isoforms could affect migration, but no empirical band size or distinct isoform bands are established.
| Band near 33.4 kDa | Consistent with predicted VGLL2 size; confirm identity with controls |
| Two bands at different positions | Could reflect isoforms 1 and 2; their migration difference is unestablished |
| Weak band in whole-cell lysate | Nuclear VGLL2 may be diluted in the sample |
| Band enriched in nuclear fraction | Consistent with VGLL2 nuclear localization |
💡Expected VGLL2 appearanceVGLL2 has a predicted mass of 33.4 kDa, but no empirical band size or resolvable migration pattern for its two isoforms is supplied; confirm band identity with appropriate controls.
| Predicted VGLL2 mass | Provides a 33.4 kDa reference, not a validated apparent band size |
| Alternative splicing | Could alter apparent size, but no migration difference is established |
| Isoform 1 | Its size relative to isoform 2 is unspecified |
| Isoform 2 | Its size relative to isoform 1 is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear VGLL2 may be poorly represented | Check a nuclear fraction and verify fraction recovery |
| Band higher than expected | Band identity or migration difference is unestablished | Confirm with VGLL2 knockdown or an independent antibody |
| Band lower than expected | Band identity or migration difference is unestablished | Confirm with VGLL2 knockdown or an independent antibody |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are unproven | Compare isoform-specific expression and VGLL2 knockdown |
| Weak or no signal | Nuclear VGLL2 may be diluted in whole-cell lysate | Enrich the nuclear fraction and check sample loading |
Experiment evidence
Western blot images showing VGLL2 band
A11724 · VGLL2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VGLL2 Western blot
🧪For positive controls for VGLL2 in Western blot, you can use no HPA-supported tissue or cell line from the supplied evidence.
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: VGLL2 is nuclear, but the supplied HPA record has no expression data to guide control selection.
HPA tissue expression evidence for VGLL2
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 VGLL2 Western Blot Tips
Deeper troubleshooting and optimisation questions for VGLL2, answered from its protein features.
How should VGLL2 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 VGLL2 isoforms produce different bands?
Isoforms · Yes. UniProt lists two isoforms; isoform 2 lacks residues 131–304 in canonical UniProt numbering. It may therefore run below isoform 1, but these features do not establish where either band appears on a blot.
Could modification explain a shifted VGLL2 band?
PTM · The supplied UniProt features list no modified residues or glycosylation sites. They provide no specific modification to test as an explanation. Even a listed modification would not, by itself, establish a visible band shift.
Does this guide establish induction of VGLL2?
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 VGLL2 Western blot?
Transfer · The canonical predicted mass is 33.4 kDa, while isoform 2 lacks residues 131–304. Choose transfer conditions that retain proteins in this size range, and check transfer of both bands if testing for both isoforms. The supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A11724 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.
What should guide VGLL2 quantitation?
Quantitation · Decide whether to quantify isoform 1, isoform 2, or both before selecting bands. Isoform 2 lacks canonical UniProt residues 131–304, so verify that the antibody recognizes both if combining their signals. VGLL2 is annotated as nuclear, which can guide sample preparation.
What size band is expected for VGLL2?
Interpretation · The canonical VGLL2 sequence is 317 amino acids with a predicted mass of 33.4 kDa. No observed band position is supplied, so use 33.4 kDa as a reference rather than an expected apparent mass.
Check the antibody's recognized sequence against the isoform difference. An epitope within canonical UniProt residues 131–304 is absent from isoform 2; an antibody targeting that region would not detect isoform 2.
Compare bands with the 33.4 kDa canonical prediction and consider the shorter isoform 2, which lacks canonical UniProt residues 131–304. Check whether the antibody can recognize that isoform. Without an observed reference band, band position alone cannot identify either isoform.
Check the antibody's recognized sequence against the isoform difference. An epitope within canonical UniProt residues 131–304 is absent from isoform 2; an antibody targeting that region would not detect isoform 2.
Compare bands with the 33.4 kDa canonical prediction and consider the shorter isoform 2, which lacks canonical UniProt residues 131–304. Check whether the antibody can recognize that isoform. Without an observed reference band, band position alone cannot identify either isoform.
Boster reagents
VGLL2 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-VGLL2 Antibody
Cat # A11724
the supplier A11724 is a polyclonal anti-VGLL2 antibody listed for human, mouse, and rat. Its WB image shows rat and mouse heart lysates and HeLa whole cell lysate, each at 40 µg, using a 1:500 antibody dilution. Evidence shown is limited to those samples and conditions.
Which to pick: A11724 is the only listed VGLL2 antibody. Its WB image provides a starting reference for rat and mouse heart tissue and HeLa cells at 40 µg per lane and 1:500 dilution; confirm performance for your own samples and conditions.
Source: BosterBio VGLL2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8N8G2.
- Human Protein Atlas. VGLL2 tissue expression.
- PMC12061476 — target-verified WB comparison
- PMC11092657 — target-verified WB comparison
- PMC7566471 — target-verified WB comparison