This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
VANGL2 / Vang-like protein 2 · Western blot design guide
Design a Western Blot for VANGL2
Real validated VANGL2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VANGL2 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
VANGL2 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 | ~59.7 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 | 1 isoform(s) |
Section 1
Real Curated VANGL2 Western Blot Protocols
The A04309 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04309) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04309) |
| Primary antibody | A04309 · 1:1000 (catalog A04309) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04309) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04309) |
Section 2
What Is the Expected VANGL2 Western Blot Band Size?
VANGL2 has a predicted monomer mass of 59.7 kDa; dimerization may affect bands, but no empirical migration or size shift is established.
| Band near 59.7 kDa | consistent with the predicted VANGL2 monomer; confirm its identity with controls |
| Band near 119 kDa | could reflect a VANGL2 homodimer if it remains intact during electrophoresis |
| Additional higher band of uncertain size | could reflect a VANGL2–VANGL1 heterodimer if the complex persists |
| Faint or absent band in lysate | membrane VANGL2 may be poorly recovered during extraction |
💡Expected VANGL2 appearanceThe predicted VANGL2 monomer is 59.7 kDa; no empirical band size is supplied, and band identity should be checked with ordinary controls.
| Predicted monomer mass | 59.7 kDa for the full-length protein; actual migration is unverified |
| VANGL2 homodimer | could appear near twice the monomer mass if it survives electrophoresis |
| VANGL1 heterodimer | could produce a higher band if intact; its apparent mass is unknown |
| Multi-pass membrane topology | may affect apparent migration, but no size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane VANGL2 may be poorly extracted | check membrane fraction recovery and use a positive control |
| Band higher than expected | a VANGL2 homodimer or VANGL1 heterodimer may persist | compare reducing and denaturing conditions and verify band identity |
| Band lower than expected | band identity or sample integrity is uncertain; no cleavage feature is supplied | check sample integrity and confirm with an independent antibody |
| Multiple bands | monomer and persistent dimers are possible, but band identities are unproven | compare denaturing conditions and use VANGL2-specific controls |
| Weak or no signal | insufficient recovery of this multi-pass membrane protein is possible | check solubilization and include a positive control |
Experiment evidence
Western blot images showing VANGL2 band
A04309 · VANGL2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VANGL2 Western blot
🧪For positive controls for VANGL2 in Western blot, you can use no HPA-supported tissue or cell sample because none was 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: VANGL2 is a multi-pass membrane protein, and the supplied HPA evidence identifies no validated positive or negative tissue.
HPA tissue expression evidence for VANGL2
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 VANGL2 Western Blot Tips
Deeper troubleshooting and optimisation questions for VANGL2, answered from its protein features.
How should VANGL2 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 VANGL2 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. The supplied features do not support assigning additional bands to annotated isoforms.
Can annotated modifications explain a shifted VANGL2 band?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore do not identify a modification that explains a shift; they also cannot establish why an observed band differs from 59.7 kDa.
Does this guide establish induction of VANGL2?
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 VANGL2 Western blot?
Transfer · VANGL2 is a 521-residue, multi-pass membrane protein with a predicted mass of 59.7 kDa. Check transfer efficiency at that size and optimize transfer conditions for recovery of membrane proteins; the supplied features do not specify a single validated method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04309 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 I quantify VANGL2 across samples?
Quantitation · Because VANGL2 is a cell-membrane protein, compare samples prepared with the same extraction method and normalize within the same sample fraction. Differences in membrane recovery could otherwise affect the measured signal.
What molecular weight should I expect for VANGL2?
Interpretation · The predicted mass is 59.7 kDa. No empirical apparent band position is supplied, so use 59.7 kDa as a reference rather than an exact migration target.
VANGL2 is located at the cell membrane and has multiple membrane-spanning regions. Consider a membrane-enriched preparation and verify that the extraction method recovers VANGL2 before comparing samples.
VANGL2 is annotated as a homodimer and as a heterodimer with VANGL1. This makes association a possibility to investigate, but the feature does not establish that a higher Western-blot band is a dimer. Compare sample preparation conditions before assigning its identity.
VANGL2 is located at the cell membrane and has multiple membrane-spanning regions. Consider a membrane-enriched preparation and verify that the extraction method recovers VANGL2 before comparing samples.
VANGL2 is annotated as a homodimer and as a heterodimer with VANGL1. This makes association a possibility to investigate, but the feature does not establish that a higher Western-blot band is a dimer. Compare sample preparation conditions before assigning its identity.
Boster reagents
VANGL2 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-Vang-like protein 2 VANGL2 Antibody
Cat # A04309
The catalog reports A04309, an anti-VANGL2 antibody with reported Human and Mouse reactivity. Its Western blot image shows extracts from various cell lines tested at 1:1000 dilution; the caption does not identify those cell lines.
Which to pick: A04309 is the only listed option. It has a Western blot image using 25 µg of cell-line extract per lane and a 1:1000 primary dilution. Check whether its reported Human or Mouse reactivity fits your sample.
Source: BosterBio VANGL2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9ULK5.
- Human Protein Atlas. VANGL2 tissue expression.
- PMC6295236 — target-verified WB comparison
- PMC11790156 — target-verified WB comparison
- PMC3282788 — target-verified WB comparison