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- Table of Contents
SNCA / Alpha-synuclein · Western blot design guide
Design a Western Blot for SNCA
Real validated SNCA Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SNCA 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
SNCA 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 | ~14.5 kDa | |
| Observed band | ~18 kDa | |
| Gel | 13% (catalog A00215-3) | |
| Positive control | Bone marrow (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated SNCA Western Blot Protocols
The A00215-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain, mouse brain (catalog A00215-3) |
| Gel % | 13% (catalog A00215-3) |
| Load | 30 ug; reducing conditions (catalog A00215-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00215-3) |
| Membrane | nitrocellulose membrane (catalog A00215-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00215-3) |
| Primary antibody | A00215-3 · 0.5 μg/mL (catalog A00215-3) |
| Primary incubation | overnight at 4°C (catalog A00215-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00215-3) |
| Secondary incubation | 1.5 hour at RT (catalog A00215-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00215-3) |
| Detection | ECL (catalog A00215-3) |
Section 2
What Is the Expected SNCA Western Blot Band Size?
SNCA is predicted at 14.5 kDa and observed at ~18 kDa in reducing brain lysates; the cause of the difference is not established.
| Band at ~18 kDa | Empirical SNCA band in reducing brain lysates; confirm identity with a positive control. |
| Band near 14.5 kDa | Near the UniProt predicted mass; band identity still needs confirmation. |
| Several bands at different positions | Could include splice isoforms 1, 2-4, and 2-5; distinct migration is unverified. |
| Close doublet | Could reflect different phosphorylation states at Ser87, Tyr125, or Ser129; verify experimentally. |
💡Expected SNCA appearanceUniProt predicts 14.5 kDa, while antibody QC shows a band at ~18 kDa in reducing brain lysates; the cause of the difference is unestablished, so confirm identity with appropriate controls.
| UniProt predicted mass | 14.5 kDa is predicted; the empirical band is ~18 kDa, with no established cause for the difference. |
| Splice isoform 1 | Its individual apparent size is not supplied. |
| Splice isoform 2-4 | Splicing may change its size; its individual migration is not supplied. |
| Splice isoform 2-5 | Splicing may change its size; its individual migration is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SNCA signal may be below detection in the sampled lysate. | Check antibody performance against a brain lysate positive control. |
| Band higher than expected | The ~18 kDa empirical band exceeds the 14.5 kDa prediction for an unestablished reason. | Compare with a validated SNCA positive control. |
| Band lower than expected | A splice isoform is possible, but its migration is unknown. | Confirm band identity with an independent SNCA antibody. |
| Multiple bands | SNCA has three named splice isoforms and documented phosphorylation sites; their band patterns are unestablished. | Use an independent antibody and, for a suspected phospho band, a phosphatase control. |
| Weak or no signal | SNCA abundance or antibody detection may be insufficient in the sample. | Run brain lysate as a positive control and check sample loading. |
Experiment evidence
Western blot images showing SNCA band
A00215-3 · SNCA
P00215-1 · SNCA
M00215-1 · SNCA
M00215 · SNCA
M00215-2 · SNCA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SNCA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SNCA in Western blot, you can use bone marrow lysate.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA lists adipose tissue as not detected, but validate it as a negative by Western blot.
HPA tissue expression evidence for SNCA
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuropil | High | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | High | Protein (IHC) | HPA → |
| Cerebellum | processes in granular layer | Medium | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SNCA Western Blot Tips
Deeper troubleshooting and optimisation questions for SNCA, answered from its protein features.
How should SNCA 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.
How could SNCA isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms: 1, 2-4 and 2-5. Isoform 2-5 lacks residues 41–54; isoform 2-4 lacks residues 103–130. Antibodies against either deleted region may miss the corresponding isoform. Check the antibody epitope before assigning separate bands to isoforms.
Isoform 2-4 lacks UniProt residues 103–130, which include phosphotyrosine 125 and phosphoserine 129. A signal from an antibody to either site therefore should not be assigned to isoform 2-4. Check isoform expression and antibody specificity when comparing phospho and total SNCA signals.
Isoform 2-4 lacks UniProt residues 103–130, which include phosphotyrosine 125 and phosphoserine 129. A signal from an antibody to either site therefore should not be assigned to isoform 2-4. Check isoform expression and antibody specificity when comparing phospho and total SNCA signals.
Which SNCA modifications matter when choosing phospho-specific antibodies?
PTM · UniProt lists phosphoserine at residue 87, phosphotyrosine at residue 125 by FYN, and phosphoserine at residue 129 by BARK1, PLK2, CK2, CK1 and GRK5. It also lists N-acetylmethionine at residue 1. These are UniProt coordinates; check the antibody’s numbering convention and target site before interpreting a signal.
Does this guide establish induction of SNCA?
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 SNCA Western blot?
Transfer · Plan transfer around the 14.5 kDa predicted protein and the supplied ~18 kDa apparent band. Check that this low-mass region is retained on the membrane, and inspect the post-transfer gel if recovery is uncertain. The supplied features do not establish one optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00215-3 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 SNCA bands be quantified across sample fractions?
Quantitation · UniProt places SNCA in the cytoplasm, membrane and nucleus and lists soluble monomers alongside homotetramers. Keep fraction preparation and detection conditions consistent across samples. Quantify equivalent bands within the same fraction and document which form or antibody signal is measured; a phospho-specific signal should not automatically stand for total SNCA.
Why might SNCA appear near 18 kDa instead of 14.5 kDa?
Interpretation · The supplied apparent band is ~18 kDa, while the predicted mass is 14.5 kDa. UniProt lists acetylation, phosphorylation and alternative isoforms, but those features alone do not explain the difference or establish a visible shift. Compare the band with an appropriate molecular weight marker and confirm its identity independently.
Consider the listed isoforms, modified residues and monomer–homotetramer population when investigating extra bands. Check band identity with an appropriate control and the antibody’s epitope. Those features are possibilities, not proof that any particular extra band is an isoform, phosphorylated protein or tetramer.
Consider the listed isoforms, modified residues and monomer–homotetramer population when investigating extra bands. Check band identity with an appropriate control and the antibody’s epitope. Those features are possibilities, not proof that any particular extra band is an isoform, phosphorylated protein or tetramer.
Boster reagents
SNCA 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-Alpha Synuclein/SNCA Antibody Picoband®
Cat # A00215-3
Real WB data
Anti-Phospho-alpha Synuclein (S129) SNCA Rabbit Monoclonal Antibody
Cat # P00215-1
Real WB data
Anti-Alpha Synuclein SNCA Rabbit Monoclonal Antibody
Cat # M00215-1
Real WB data
Anti-alpha + beta Synuclein SNCA Rabbit Monoclonal Antibody
Cat # M00215
Real WB data
Anti-SNCA Antibody (C-term)
Cat # M00215-2
Real WB data
Anti-Phospho-alpha Synuclein (S129) SNCA Rabbit Monoclonal Antibody
Cat # P00215-2
Five listed antibodies have WB images: SNCA in rat and mouse brain, human cell lines, or human brain tissue; phospho-SNCA (S129) in 293T lysates; and alpha/beta synuclein in rodent brain. These captions document specific samples and conditions, not broader validation.
Which to pick: For rat or mouse brain, consider A00215-3 for SNCA or M00215 for alpha/beta synuclein. For human samples, M00215-1 shows cell lysates and M00215-2 shows brain tissue. Choose P00215-1 when measuring S129 phosphorylation; its image uses 293T lysates.
Source: BosterBio SNCA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P37840.
- Human Protein Atlas. SNCA tissue expression.
- PMC12398189 — target-verified WB comparison
- PMC3918138 — target-verified WB comparison
- PMC12758206 — target-verified WB comparison