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- Table of Contents
VAMP8 / Vesicle-associated membrane protein 8 · Western blot design guide
Design a Western Blot for VAMP8
Real validated VAMP8 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-VAMP8 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
VAMP8 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 | ~11.4 kDa | |
| Observed band | ~11 kDa | |
| Gel | 5–20% (catalog M02338) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated VAMP8 Western Blot Protocols
The M02338 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human THP-1, human RT4, human MCF-7 (catalog M02338) |
| Gel % | 5–20% (catalog M02338) |
| Load | 30 ug; reducing conditions (catalog M02338) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M02338) |
| Membrane | nitrocellulose membrane (catalog M02338) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M02338) |
| Primary antibody | M02338 · 1:500 (catalog M02338) |
| Primary incubation | overnight at 4°C (catalog M02338) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M02338) |
| Secondary incubation | 1.5 hour at RT (catalog M02338) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M02338) |
| Detection | ECL (catalog M02338) |
Section 2
What Is the Expected VAMP8 Western Blot Band Size?
VAMP8 is predicted at 11.4 kDa and observed at ~11 kDa; the cause of the small difference is not established.
| Band at ~11 kDa | matches the empirical VAMP8 band |
| Faint ~11 kDa band in whole-cell lysate | VAMP8 is a membrane protein and may be poorly recovered |
| No ~11 kDa band in whole-cell lysate | membrane-associated VAMP8 may be underrepresented in the sample |
| Close doublet near ~11 kDa | may reflect different phosphorylation states, but distinct migration is unproven |
💡Expected VAMP8 appearanceUniProt predicts 11.4 kDa, and antibody QC shows a band at ~11 kDa; confirm its identity with an appropriate positive sample and a VAMP8 depletion control.
| UniProt predicted mass | places the unmodified protein near 11.4 kDa, consistent with the observed ~11 kDa band |
| N-acetylmethionine at residue 1 | is documented, with no demonstrated visible size shift |
| Phosphoserine at residues 5, 18 and 55 | may alter mobility, but no distinct band shift is established |
| Phosphothreonine at residues 28, 48 and 54 | may alter mobility, but no distinct band shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated VAMP8 may be poorly recovered. | Check membrane extraction and test a membrane-enriched sample. |
| Band higher than expected | The band's identity or mobility is uncertain; phosphorylation sites alone do not establish the shift. | Compare with the ~11 kDa band and test VAMP8-depleted lysate. |
| Band lower than expected | The band may be a degradation product or unrelated signal. | Use fresh, protease-inhibited lysate and test VAMP8 depletion. |
| Multiple bands | Phosphorylation states are possible, but distinct migration is unproven. | Compare phosphatase-treated and untreated samples, then confirm bands by VAMP8 depletion. |
| Weak or no signal | Recovery of membrane-associated VAMP8 may be low. | Check extraction and loading with a positive sample. |
| Fragments below expected size | Protein degradation is possible. | Prepare fresh lysate with protease inhibitors and confirm fragment identity by VAMP8 depletion. |
Experiment evidence
Western blot images showing VAMP8 band
M02338 · VAMP8
M02338 · VAMP8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for VAMP8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for VAMP8 in Western blot, you can use appendix tissue, which HPA scores High.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: A not-detected tissue is available, but VAMP8’s membrane location may make efficient extraction important.
HPA tissue expression evidence for VAMP8
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 |
|---|---|---|---|---|
| Appendix | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced VAMP8 Western Blot Tips
Deeper troubleshooting and optimisation questions for VAMP8, answered from its protein features.
How should VAMP8 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 VAMP8 isoforms explain multiple bands?
Isoforms · The supplied UniProt features list one isoform and no alternative sequence. They provide no isoform-based explanation for multiple bands.
Which VAMP8 modifications matter when interpreting a band?
PTM · Using the supplied UniProt coordinates, VAMP8 has N-acetylmethionine at position 1; phosphoserine at 5, 18 and 55; and phosphothreonine at 28, 48 and 54. Check the antibody's numbering convention before comparing sites. These features alone do not establish a visible band shift.
Does this guide establish induction of VAMP8?
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 VAMP8 Western blot?
Transfer · VAMP8 is a small, 100-residue, single-pass membrane protein with a predicted mass of 11.4 kDa. Optimize transfer for a small membrane protein, then check the membrane and post-transfer gel to assess retention and transfer. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02338 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 VAMP8 bands be quantified?
Quantitation · Quantify the ~11 kDa band consistently across samples, using exposures within the linear detection range and an appropriate loading reference. Because VAMP8 is listed at endosome and lysosome membranes, keep the sampled fraction consistent when comparing abundance.
Should VAMP8 migrate at its predicted mass?
Interpretation · VAMP8 is predicted at 11.4 kDa, close to the observed ~11 kDa band. Its listed modifications do not establish a visible shift or explain a mass difference by themselves.
First compare them with the expected ~11 kDa band. VAMP8 participates in SNARE complexes, but the supplied features do not establish that a particular extra band is a complex, modification or isoform. Check whether the band follows VAMP8 across samples and validate its identity independently.
First compare them with the expected ~11 kDa band. VAMP8 participates in SNARE complexes, but the supplied features do not establish that a particular extra band is a complex, modification or isoform. Check whether the band follows VAMP8 across samples and validate its identity independently.
Boster reagents
VAMP8 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-VAMP8/Endobrevin Rabbit Monoclonal Antibody
Cat # M02338
The catalog reports one anti-VAMP8 antibody for Western blot: rabbit monoclonal M02338. Its WB image shows an approximately 11 kDa band, matching the expected size, in the lysates named in the caption. The supplied evidence is a product image; no orthogonal validation is provided.
Which to pick: M02338 is the only listed option. It reports human, mouse, and rat reactivity and has a WB image using human cell, rat cell, and mouse cell and lung lysates. Match your sample to those tested contexts and the reported conditions.
Source: BosterBio VAMP8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9BV40.
- Human Protein Atlas. VAMP8 tissue expression.
- PMC12505561 — target-verified WB comparison
- PMC9605769 — target-verified WB comparison
- PMC11680461 — target-verified WB comparison