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SOX10 · Western blot design guide
Design a Western Blot for SOX10
Real validated SOX10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SOX10 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Last reviewed: May 2026 · Scientific review: Boster Bio technical team
SOX10 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 | ~49.9 kDa | |
| Observed band | 60-65 kDa | |
| Gel | 10–12% | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-induced mobility shift | |
| Regulation | Kras signaling dn | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SOX10 Western Blot Protocols
Literature-validated Western blot parameters for SOX10 — gel percentage, transfer, blocking, antibody incubation and detection, extracted from published methods.
| Sample / lysate | human SH-SY5Y , Lane 2: human U251 , Lane 3: human U2OS , Lane 4: human U87 , Lane 5: rat brain , Lane 6: rat C6 , Lane 7: mouse brain , Lane 8: mouse Neuro-2a . After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SOX10 antigen affinity purified polyclonal antibody (Catalog # A00758-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SOX10 at approximately 60 kDa. The expected band size for SOX10 is at 50 kDa |
| Gel % | 10–12% |
| Load | 30 ug |
| Transfer | nitrocellulose membrane, 150 mA, 50–90 min |
| Membrane | nitrocellulose |
| Blocking | 5% non-fat milk / TBS, 1.5 h RT |
| Primary antibody | 0.5 µg/mL |
| Primary incubation | overnight at 4 °C |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 |
| Wash | TBS-0.1% Tween, 3 × 5 min |
| Detection | ECL |
| Exposure / imaging | Tanon 5200 |
| Observed band | 60 kDa |
Section 2
What Is the Expected SOX10 Western Blot Band Size?
SOX10 has a 49.9 kDa calculated backbone but runs at 60-65 kDa on Western blot, an anomalous migration shift typical of basic DNA-binding transcription factors.
| Single band around 60-65 kDa | the native SOX10 signal, running well above the 49.9 kDa calculated mass due to anomalous SDS-PAGE migration typical of this basic DNA-binding transcription factor rather than a modification |
| Faint band near 50 kDa | position matching the unmodified calculated mass, sometimes seen when the protein migrates closer to its true mass rather than showing the usual anomalous shift |
| Two closely spaced bands | reflects expression of both annotated splice isoforms (1 and 2) rather than degradation or nonspecific binding |
| Signal detected across nuclear and cytoplasmic lysate fractions | consistent with SOX10 shuttling between cytoplasm, nucleus, and the mitochondrial outer membrane rather than being confined to one compartment |
| No shift in band position between reducing and non-reducing sample buffer | expected because SOX10 has no annotated disulfide bonds or disulfide-linked dimer formation |
| Clean single band with no high-molecular-weight smear | expected because SOX10 has no annotated glycosylation sites that would create heterogeneous glycoform smearing |
💡Expected SOX10 appearanceExpect a single dominant SOX10 band at 60-65 kDa, well above the 49.9 kDa calculated mass, reflecting anomalous SDS-PAGE migration typical of this basic DNA-binding transcription factor rather than a modification-driven shift.
| Predicted mass from UniProt sequence | 466 aa backbone calculates to 49.9 kDa, the theoretical baseline before any migration effects |
| DNA-binding transcription factor domain | the basic, charged DNA-binding/transcriptional regulator domain binds SDS atypically, causing anomalous slower migration and an apparent size well above the calculated mass |
| Serine-24 phosphorylation | adds only a small amount of mass and contributes at most marginally to any upward shift, not the full gap seen on blot |
| Alternative splicing, isoforms 1 and 2 | the isoform expressed in a given sample can shift which band position or relative intensity predominates |
| Absence of glycosylation and disulfide bonding | rules out glycosylation-driven smearing and disulfide-linked dimerization as causes of the observed band position |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | anomalous SDS-PAGE migration of the basic DNA-binding domain, not degradation or error | compare against the established 60-65 kDa reference band and confirm with a known-positive lysate before troubleshooting further |
| Multiple bands | co-expression of the two annotated splice isoforms | check which isoform is expressed in the tissue or cell line and whether the antibody epitope is common to both |
| Band lower than expected | proteolytic degradation during lysate preparation, since no cleavage event is annotated to naturally produce a smaller form | add protease inhibitors, keep lysates cold throughout, and use fresh rather than freeze-thawed sample |
| Weak or no signal | SOX10 expression is lineage-restricted to neural crest derivatives such as melanocytes, Schwann cells, and oligodendrocytes | use a characterized positive-control lysate from a relevant lineage and increase total protein loaded |
| Broad smear instead of sharp band | aggregation or incomplete denaturation of this nuclear DNA-binding protein rather than glycosylation, since none is annotated | ensure complete lysis and denaturation with SDS sample buffer and adequate heating, and avoid overloading the gel |
Experiment evidence
Western blot images showing SOX10 band
A00758-1 · SOX10
A00758-3 · SOX10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SOX10 Western blot
🧪For positive controls for SOX10 in Western blot, you can use caudate tissue lysate, which shows high SOX10 expression in HPA data.
Positive control: Caudate (brain)
Negative control: Adipose tissue
Loading controls: Run GAPDH and β-actin as loading controls alongside a total-protein stain (e.g., stain-free gel, Ponceau S, or REVERT) to confirm equal loading and transfer.
⚠️Feasibility: SOX10 is a nuclear/cytoplasmic transcription factor with tissue-restricted expression, so a clean negative control (e.g., adipose tissue) is readily available, making good positive/negative control selection feasible.
HPA tissue expression evidence for SOX10
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Hippocampus | glial cells | High | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - nucleus | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SOX10 Western Blot Tips
Deeper troubleshooting and optimisation questions for SOX10, answered from its protein features.
Why does SOX10 run heavier than its predicted mass?
SOX10 has a calculated mass of 49.9 kDa but typically appears at 60-65 kDa on blots. This shift is common for DNA-binding transcription factors with acidic/proline-rich regions that reduce SDS binding and slow SDS-PAGE migration; the single reported phosphosite may also contribute modestly to apparent mass. Treat 60-65 kDa as the expected band, not the calculated 49.9 kDa.
Does SOX10 have isoforms visible on Western blot?
UniProt lists two SOX10 isoforms (1 and 2) generated by alternative splicing. If your antibody's epitope lies in a region absent from isoform 2, only isoform 1 will be detected; epitopes in shared exons will detect both, potentially as two close bands near 60-65 kDa. Check the immunogen sequence against both isoform sequences before interpreting band number.
Is SOX10 phosphorylation relevant to blot interpretation?
SOX10 carries one annotated modified residue consistent with regulation as a transcriptional activator. Phosphorylation can cause a slight upward mobility shift or doublet near the main 60-65 kDa band. Phosphatase treatment of lysate can help confirm whether a doublet reflects phosphorylation state rather than isoform expression or protein degradation.
How should blocking be optimised for SOX10 detection?
SOX10 is a low-abundance nuclear transcription factor, so use 5% non-fat milk or BSA blocking to minimize background while preserving signal; avoid over-blocking, which can mask the specific 60-65 kDa band. Since SOX10 is not glycosylated, milk-based blocking is safe and will not introduce lectin-type cross-reactivity artifacts.
What transfer method to use for SOX10 Western blot?
SOX10 runs at 60-65 kDa, a mid-range size well suited to standard wet or semi-dry transfer. Use wet transfer with 20% methanol transfer buffer for efficient transfer of this soluble nuclear/cytoplasmic protein; standard 60-90 minute transfer times at 100V are sufficient given its moderate molecular weight and monomeric state.
What loading control suits SOX10 quantitation?
Because SOX10 localizes to cytoplasm, nucleus, and the mitochondrial outer membrane, choose a loading control matching your extraction method: histone H3 or Lamin B1 for nuclear fractions, GAPDH for whole-cell lysate. Mismatched controls can misrepresent SOX10 quantitation given its multi-compartment distribution and non-secreted, monomeric nature.
What explains extra bands beyond the SOX10 band?
Additional bands can arise from the second SOX10 isoform, incomplete denaturation of this DNA-binding protein, or partial degradation during nuclear extraction. Since SOX10 has no disulfide bonds or glycosylation, unexpected higher-MW bands are unlikely to reflect aggregation via these mechanisms; verify with reducing conditions and fresh lysate preparation instead.
Boster reagents
Best SOX10 Western Blot Antibodies
BosterBio's SOX10 antibodies are among the best-performing WB antibodies on the market — well cited, thoroughly validated, and orthogonally cross-validated against negative tissues and complementary methods.
Real WB data
Anti-SOX10 Antibody Picoband®
Cat # A00758-1
Real WB data
Anti-SOX10 Antibody Picoband®
Cat # A00758-3
Our recommended anti-SOX10 antibodies are top-performing, extensively cited reagents rigorously validated by Western blot and cross-checked against negative-control tissues and orthogonal detection methods, giving confidence in specific, reproducible SOX10 detection for your experiments.
Which to pick: Both A00758-1 and A00758-3 have a real Western blot validation image confirming SOX10 detection, so either is a solid choice; no other distinguishing details are listed for these two SKUs, so pick either, or run both in parallel if you want extra cross-checking confidence.
Source: BosterBio SOX10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P56693.
- Human Protein Atlas. SOX10 tissue expression.