This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
VAMP2 / Vesicle-associated membrane protein 2 · Western blot design guide
Design a Western Blot for VAMP2
Real validated VAMP2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VAMP2 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
VAMP2 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 | ~12.7 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
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 | 1 isoform(s) |
Section 1
Real Curated VAMP2 Western Blot Protocols
The M02331-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse brain lysate (catalog M02331-1) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02331-1; 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 VAMP2 Western Blot Band Size?
VAMP2 is predicted at 12.7 kDa; membrane localization and residue 2 acetylation are documented, but neither establishes a different migration position.
| Band near 12.7 kDa | consistent with the predicted VAMP2 size; confirm identity with antibody controls |
| Weak or absent band in soluble lysate | VAMP2 is a vesicle membrane protein and may be underrepresented in this fraction |
| Stronger band in a vesicle membrane fraction | consistent with VAMP2 localization to synaptic vesicle membranes |
| No distinct acetylation-related doublet | N-acetylserine at residue 2 does not establish a resolvable shift |
💡Expected VAMP2 appearanceUniProt predicts VAMP2 at 12.7 kDa, but no empirical band size is supplied; confirm a candidate band with antibody specificity controls and appropriate membrane-containing samples.
| UniProt predicted mass | sets a 12.7 kDa reference, not a measured migration position |
| 116-amino-acid sequence | underlies the predicted mass; no separate size shift is established |
| N-acetylserine at residue 2 | is documented, but no resolvable band shift is established |
| Single-pass type IV membrane localization | affects recovery from samples; no apparent size shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | VAMP2 may be underrepresented if vesicle membranes are poorly recovered | check a membrane-containing fraction and sample preparation |
| Band higher than expected | the supplied features do not establish an upward migration shift | check antibody specificity and compare with a VAMP2-positive sample |
| Band lower than expected | a lower band has no established feature-based assignment | check antibody specificity and sample integrity |
| Multiple bands | the single listed isoform does not establish distinct VAMP2 bands | compare positive and negative controls to identify the specific band |
| Weak or no signal | low recovery of vesicle membrane material may reduce VAMP2 signal | assess membrane recovery and use a VAMP2-positive control |
Experiment evidence
Western blot images showing VAMP2 band
M02331-1 · VAMP2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VAMP2 Western blot
🧪For positive controls for VAMP2 in Western blot, you can use an HPA-IHC candidate positive sample once one is identified.
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 provides no tissue data, so positive and negative tissue controls cannot be selected from this evidence.
HPA tissue expression evidence for VAMP2
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 VAMP2 Western Blot Tips
Deeper troubleshooting and optimisation questions for VAMP2, answered from its protein features.
How should VAMP2 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 VAMP2 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Does a listed VAMP2 modification predict a band shift?
PTM · UniProt lists N-acetylserine at position 2, using UniProt numbering. It does not establish a visible band shift. The phosphoprotein keyword supplies no phosphorylation site or condition here.
Does this guide establish induction of VAMP2?
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 VAMP2 Western blot?
Transfer · For this 12.7 kDa protein, consider a 0.2 µm membrane and check that the protein is retained after transfer. VAMP2 is a single-pass membrane protein, so also verify that sample preparation extracts it effectively.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02331-1 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 VAMP2 bands be quantified?
Quantitation · Measure bands within the assay’s linear range and normalize against a consistent loading measure. Because VAMP2 is associated with synaptic vesicle membranes, keep membrane extraction conditions consistent across samples.
Should VAMP2 run exactly at its predicted 12.7 kDa?
Interpretation · 12.7 kDa is the predicted mass; no observed band position is supplied. The listed features alone cannot establish a visible shift or explain a difference from the predicted mass. Compare the band with a suitable low-mass marker.
VAMP2 is part of a SNARE complex with SNAP25 and STX1A, but that annotation alone cannot identify an extra band as a complex. Check band specificity and sample preparation before assigning an identity. No empirical band position is supplied.
VAMP2 is part of a SNARE complex with SNAP25 and STX1A, but that annotation alone cannot identify an extra band as a complex. Check band specificity and sample preparation before assigning an identity. No empirical band position is supplied.
Boster reagents
VAMP2 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-VAMP2 Rabbit Monoclonal Antibody
Cat # M02331-1
The catalog reports one anti-VAMP2 rabbit monoclonal antibody for Western blotting. Its reported reactivity is human, mouse, and rat; the supplied WB image shows VAMP2 expression in mouse brain lysate. Other sample contexts are not shown.
Which to pick: M02331-1 is the only listed option. Choose it when its reported reactivity fits your sample; the supplied WB image documents mouse brain lysate, so assess other samples with appropriate controls.
Source: BosterBio VAMP2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P63027.
- Human Protein Atlas. VAMP2 tissue expression.
- PMC3151102 — target-verified WB comparison
- PMC5880409 — target-verified WB comparison
- PMC4156046 — target-verified WB comparison