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- Table of Contents
SCN8A / Sodium channel protein type 8 subunit alpha · Western blot design guide
Design a Western Blot for SCN8A
Source-linked SCN8A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SCN8A 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
SCN8A Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~225.3 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked SCN8A Western Blot Protocol Options
The A02015 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF-7, Raw264.7, H9C2 (catalog A02015) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02015; 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 SCN8A Western Blot Band Size?
SCN8A is predicted at 225.3 kDa; glycosylation, disulfide linkage, and isoforms could alter its band pattern, but no empirical migration is supplied.
| Band near 225.3 kDa | consistent with the predicted full-length SCN8A mass; confirm identity with controls |
| Band above 225.3 kDa | N-linked glycosylation could affect migration |
| Band near twice the monomer size under non-reducing conditions | consistent with an inter-chain disulfide at Cys904 |
| Broad band or smear | variable N-linked glycosylation is possible but unconfirmed |
| Several bands at different positions | isoforms 1 through 5 are possible sources, but distinct migration is unestablished |
💡Expected SCN8A appearanceSCN8A has a predicted mass of 225.3 kDa, but no empirical band size is supplied; assess bands near or above that size with reduction, glycosidase, and antibody-specificity controls.
| Predicted SCN8A mass | 225.3 kDa is the sequence-based reference, not a measured band |
| N-linked glycosylation at Asn215, Asn289, Asn295, and Asn308 | could increase apparent size if these sites are occupied |
| High-mannose N-linked glycosylation at Asn326 | could affect migration if occupied |
| N-linked glycosylation at Asn1358, Asn1372, and Asn1383 | could increase apparent size if these sites are occupied |
| Inter-chain disulfide at Cys904 | could retain a species near twice the monomer size without complete reduction |
| Splice isoforms 1, 2, 3, 4, and 5 | may differ in size, but their masses and separation are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SCN8A is a multi-pass membrane protein that may be poorly recovered | check membrane enrichment and solubilization, then confirm antibody specificity |
| Band higher than expected | incomplete reduction may retain a disulfide-linked species, or N-linked glycans may affect migration | compare reducing and non-reducing lanes and test glycosidase sensitivity |
| Band lower than expected | a splice isoform is possible, but its mass is unknown | check antibody epitope coverage and verify band identity with an independent antibody or SCN8A depletion |
| Broad smear instead of sharp band | variation in N-linked glycosylation is possible | compare treated and untreated samples with a deglycosylation control |
| Multiple bands | splice isoforms are possible, though distinct bands are not established | compare antibody epitopes and confirm candidate bands by SCN8A depletion |
Experiment evidence
Western blot images showing SCN8A band
A02015 · SCN8A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SCN8A Western blot
🧪For positive controls for SCN8A in Western blot, you can use no HPA-supported positive sample from the supplied evidence.
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 identifies no tissue controls, and SCN8A is a multi-pass membrane protein.
HPA tissue expression evidence for SCN8A
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 SCN8A Western Blot Tips
Deeper troubleshooting and optimisation questions for SCN8A, answered from its protein features.
How should SCN8A 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.
Which SCN8A isoforms could produce different bands?
Isoforms · Five isoforms are listed. Isoform 4 lacks reference-sequence residues 1284–1980, and isoform 5 lacks 1275–1315; these deletions could affect band size. Isoform 3 changes residue 666 from E to EVKIDKAATDDS, while isoform 2 has substitutions at 207 and 212. These are UniProt reference-sequence coordinates; band positions are not supplied.
Which SCN8A phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserines at reference-sequence positions 518 and 520, plus phosphoserine 1497 modified by PKC. If comparing phosphorylation-sensitive signals, account for these annotated sites. Their presence alone does not predict a visible mobility shift; antibody or paper numbering may use a different sequence convention.
UniProt lists N-linked sites at reference-sequence residues 215, 289, 295, 308, 326, 1358, 1372 and 1383. Residue 326 is annotated as high mannose. Keep these coordinates explicit when interpreting a glycosylation-related band change; the annotations alone do not establish its size or cause.
UniProt lists N-linked sites at reference-sequence residues 215, 289, 295, 308, 326, 1358, 1372 and 1383. Residue 326 is annotated as high mannose. Keep these coordinates explicit when interpreting a glycosylation-related band change; the annotations alone do not establish its size or cause.
Does this guide establish induction of SCN8A?
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 SCN8A Western blot?
Transfer · SCN8A is a 225.3 kDa multi-pass membrane protein. Start with wet transfer suited to high-mass proteins, then check the membrane and post-transfer gel around 225 kDa. Adjust transfer conditions if substantial protein remains in the gel.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02015 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 SCN8A bands be quantified across samples?
Quantitation · Quantify the same resolved band or band set consistently across samples and use signals within the assay's linear range. SCN8A has five annotated isoforms, so state which bands are included. Its membrane localization also makes consistent sample preparation important for comparisons.
Should SCN8A run exactly at its predicted 225.3 kDa?
Interpretation · No observed band size is supplied. SCN8A has eight annotated N-linked glycosylation sites and three phosphoserines, but these features alone do not establish a visible shift or explain a measured mass difference. Compare the observed band with 225.3 kDa while keeping apparent and predicted mass distinct.
Consider the annotated isoforms, especially the large C-terminal deletion in isoform 4 and the 41-residue deletion in isoform 5. SCN8A also has eight N-linked glycosylation sites and three phosphoserines. These features suggest possibilities, but cannot identify an unexpected band without additional evidence.
Consider the annotated isoforms, especially the large C-terminal deletion in isoform 4 and the 41-residue deletion in isoform 5. SCN8A also has eight N-linked glycosylation sites and three phosphoserines. These features suggest possibilities, but cannot identify an unexpected band without additional evidence.
Boster reagents
SCN8A 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-Na+ CP type VIII Alpha SCN8A Antibody
Cat # A02015
The catalog reports one anti-SCN8A polyclonal antibody, A02015, with reported Human, Mouse, and Rat reactivity. Its WB image shows testing at 1:500 in MCF-7, Raw264.7, and H9C2 lysates; these examples do not establish performance in every sample.
Which to pick: A02015 is the only listed option and has a WB image. Use the reported reactivity and the 1:500 blot in MCF-7, Raw264.7, and H9C2 lysates to judge how closely its tested conditions match yours.
Source: BosterBio SCN8A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UQD0.
- Human Protein Atlas. SCN8A tissue expression.
- PMC4077284 — target-verified WB comparison
- PMC11304301 — target-verified WB comparison
- PMC5760706 — target-verified WB comparison