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- Table of Contents
SNAP29 / Synaptosomal-associated protein 29 · Western blot design guide
Design a Western Blot for SNAP29
Real validated SNAP29 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SNAP29 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
SNAP29 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 | ~29 kDa | |
| Observed band | ~29 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SNAP29 Western Blot Protocols
The M05318 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (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 | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M05318 · 1:50 (catalog M05318) |
| 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 SNAP29 Western Blot Band Size?
SNAP29 is predicted at 29 kDa and observed at ~29 kDa; phosphorylation is documented, but a migration effect is not established.
| Band at ~29 kDa | Matches the predicted mass and the band observed in reducing whole-cell blots |
| Band slightly above ~29 kDa | May reflect phosphorylation, though a mobility shift has not been demonstrated |
| Close doublet near ~29 kDa | Could reflect different phosphorylation states; distinct bands are not established |
| Weak band in a membrane-depleted fraction | Some SNAP29 may partition with peripheral membrane compartments |
💡Expected SNAP29 appearanceSNAP29 has a predicted mass of 29 kDa and an observed band at ~29 kDa in reducing whole-cell blots; confirm band identity with an independent antibody or SNAP29 depletion.
| Predicted SNAP29 mass | Places the expected band near 29 kDa, consistent with the observed ~29 kDa band |
| Phosphoserine at Ser77 | Could alter mobility; no visible shift is established |
| Phosphoserine at Ser78 | Could alter mobility; no visible shift is established |
| Phosphothreonine at Thr130 | Could alter mobility; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SNAP29 may be lost during preparation of its cytoplasmic or peripheral membrane pools | Check extraction and fractionation, then test a validated positive lysate |
| Band higher than expected | Phosphorylation could affect mobility, but the identity of a higher band is unproven | Compare phosphatase-treated samples and confirm with SNAP29 depletion |
| Band lower than expected | No signal peptide or propeptide supports a smaller mature product | Check sample integrity and confirm the band with an independent antibody |
| Multiple bands | Phosphorylation states are possible, but distinct SNAP29 bands are unproven | Compare phosphatase treatment and SNAP29-depleted lysate |
| Weak or no signal | Incomplete recovery of cytoplasmic or peripheral membrane SNAP29 may reduce signal | Check fraction recovery and run a validated positive lysate |
Experiment evidence
Western blot images showing SNAP29 band
M05318 · SNAP29
M05318 · SNAP29
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SNAP29 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SNAP29 in Western blot, you can use appendix lysate, which has medium HPA expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA reports only medium signal in appendix, so the positive control may require careful detection.
HPA tissue expression evidence for SNAP29
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 | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | adipocytes | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | molecular layer - neuropil | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SNAP29 Western Blot Tips
Deeper troubleshooting and optimisation questions for SNAP29, answered from its protein features.
How should SNAP29 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 SNAP29 isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. These features do not support assigning additional bands to SNAP29 isoforms; check their specificity before interpreting them.
Which phosphorylation sites are annotated for SNAP29?
PTM · UniProt lists phosphoserine at positions 77, 78, 114, 163, 182, 185, 204 and 210, and phosphothreonine at 130 and 137. Use these UniProt coordinates when selecting site-specific reagents; numbering in papers or antibody descriptions may differ.
Measure total SNAP29 alongside any site-specific phospho signal, then compare the phospho signal relative to total SNAP29. The ten annotated sites identify possible measurements but do not establish that every site is phosphorylated in your sample.
Measure total SNAP29 alongside any site-specific phospho signal, then compare the phospho signal relative to total SNAP29. The ten annotated sites identify possible measurements but do not establish that every site is phosphorylated in your sample.
Does this guide establish induction of SNAP29?
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.
How should transfer be checked for SNAP29?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M05318 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 SNAP29 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Should SNAP29 migrate at its predicted 29 kDa?
Interpretation · The supplied observed band is approximately 29 kDa, matching the predicted mass. Annotated phosphorylation alone does not establish a visible shift or explain a different apparent mass.
SNAP29 is annotated in the cytoplasm and as a peripheral protein on Golgi and autophagosome membranes. If fractionating samples, examine both cytosolic and membrane-associated fractions so a change in distribution is not mistaken for a change in total abundance.
SNAP29 forms a SNARE complex with VAMP8 and STX17, but that annotation does not establish that an upper Western blot band is the complex. Check band specificity and whether the sample preparation preserves complexes before making that assignment.
SNAP29 is annotated in the cytoplasm and as a peripheral protein on Golgi and autophagosome membranes. If fractionating samples, examine both cytosolic and membrane-associated fractions so a change in distribution is not mistaken for a change in total abundance.
SNAP29 forms a SNARE complex with VAMP8 and STX17, but that annotation does not establish that an upper Western blot band is the complex. Check band specificity and whether the sample preparation preserves complexes before making that assignment.
Boster reagents
SNAP29 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-SNAP29 Rabbit Monoclonal Antibody
Cat # M05318
M05318 is a rabbit monoclonal anti-SNAP29 antibody listed for human, mouse, and rat. It has a WB image; the supplied caption describes immunoprecipitation at 1:50 followed by WB at 1:3,000. Evidence for standalone WB performance is limited.
Which to pick: M05318 is the only listed option. Its stated reactivity covers human, mouse, and rat; check whether the pictured IP and WB workflow matches your planned experiment.
Source: BosterBio SNAP29 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95721.
- Human Protein Atlas. SNAP29 tissue expression.
- PMC8157373 — target-verified WB comparison