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- Table of Contents
MYO6 / Unconventional myosin-VI · Western blot design guide
Design a Western Blot for MYO6
Real validated MYO6 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYO6 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
MYO6 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 | ~149.7 kDa | |
| Observed band | ~150 kDa | |
| Gel | 5–20% (catalog A02627-2) | |
| Positive control | Bronchus (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 | 6 isoform(s) |
Section 1
Real Curated MYO6 Western Blot Protocols
The A02627-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A431, mouse brain (catalog A02627-2) |
| Gel % | 5–20% (catalog A02627-2) |
| Load | 30 ug; reducing conditions (catalog A02627-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02627-2) |
| Membrane | nitrocellulose membrane (catalog A02627-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02627-2) |
| Primary antibody | A02627-2 · 0.5 μg/mL (catalog A02627-2) |
| Primary incubation | overnight at 4°C (catalog A02627-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02627-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02627-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02627-2) |
| Detection | ECL (catalog A02627-2) |
Section 2
What Is the Expected MYO6 Western Blot Band Size?
MYO6 is predicted at 149.7 kDa and observed near 150 kDa; the small difference has no established cause in the supplied evidence.
| Band near 150 kDa | Matches the empirical MYO6 band and its 149.7 kDa predicted mass; confirm identity with controls |
| Band near twice the monomer size | Could represent a retained MYO6 homodimer if it survives SDS-PAGE |
| Several bands at different positions | Could reflect MYO6 splice isoforms; distinct migration is not established |
| Close doublet near 150 kDa | Could reflect phosphorylation at documented sites; a visible shift is not established |
💡Expected MYO6 appearanceMYO6 has a predicted mass of 149.7 kDa and an empirical band near 150 kDa in reducing whole-cell and tissue lysates; verify band identity with appropriate controls.
| Predicted MYO6 mass | 149.7 kDa, consistent with the observed band near 150 kDa |
| Homodimer formation | Could yield a band near twice the monomer size if the dimer survives SDS-PAGE |
| Splice isoforms 1, 2, and 3 | May differ in apparent size; their individual migration is not supplied |
| Splice isoforms 4, 5, and 6 | May differ in apparent size; their individual migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | MYO6 homodimer may persist during electrophoresis | Compare fully reducing and nonreducing preparations |
| Band lower than expected | A splice isoform or sample degradation is possible | Check sample integrity and confirm identity with a second MYO6 antibody |
| Multiple bands | MYO6 has six splice isoforms, though distinct migration is unproven | Compare bands with a second MYO6 antibody and an appropriate positive control |
| Weak or no signal | MYO6 abundance or antibody recognition may be low in the sample | Check transfer and compare with A431 or mouse brain positive lysate |
| Fragments below expected size | MYO6 may have degraded during sample preparation | Prepare fresh lysate with protease inhibitors and confirm band identity |
| Broad smear instead of sharp band | Sample degradation or excess loaded protein is possible | Check lysate integrity and repeat with less protein |
Experiment evidence
Western blot images showing MYO6 band
A02627-2 · MYO6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYO6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYO6 in Western blot, you can use bronchus tissue lysate, which HPA rates High.
Positive control: Bronchus (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: Tissue controls are feasible because HPA rates bronchus High and adipose tissue Not detected.
HPA tissue expression evidence for MYO6
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 |
|---|---|---|---|---|
| Bronchus | ciliated cells (cell body) | High | Protein (IHC) | HPA → |
| Cerebellum | processes in white matter | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYO6 Western Blot Tips
Deeper troubleshooting and optimisation questions for MYO6, answered from its protein features.
How should MYO6 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 MYO6 isoforms produce nearby bands?
Isoforms · Six isoforms are listed. Isoforms 2 and 5 lack residues 1037–1068; isoform 1 lacks 1037–1045; isoforms 4 and 5 lack 1147–1155; and isoform 6 replaces 1147–1156 with A. These differences could affect migration, but the features do not predict resolvable bands.
No. The listed sequence changes remove residue 1155 in isoforms 4 and 5 and replace the 1147–1156 segment with A in isoform 6. Check which isoform an assay detects before interpreting a signal assigned to canonical phosphoserine 1155.
No. The listed sequence changes remove residue 1155 in isoforms 4 and 5 and replace the 1147–1156 segment with A in isoform 6. Check which isoform an assay detects before interpreting a signal assigned to canonical phosphoserine 1155.
Which phosphorylation sites matter when interpreting MYO6 bands?
PTM · The supplied UniProt coordinates list phosphoserines 267, 604, 1025 and 1155, plus phosphothreonine 405. These are canonical sequence coordinates; antibody or paper numbering may differ. Phosphorylation alone does not establish a visible band shift.
Does this guide establish induction of MYO6?
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 MYO6 Western blot?
Transfer · MYO6 is predicted at 149.7 kDa. Choose transfer conditions suitable for a ~150 kDa protein and check that protein at this size reaches and remains on the membrane. The supplied features do not establish one preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02627-2 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 MYO6 bands be quantified across samples?
Quantitation · Quantify a consistently defined ~150 kDa band or band group, and apply the same boundary across samples. Six listed isoforms and five canonical phosphorylation sites make it useful to record which bands were included; these features alone do not identify each band.
Why does MYO6 appear near 150 kDa?
Interpretation · The supplied apparent band is ~150 kDa, close to the 149.7 kDa predicted mass. The listed phosphorylation sites and splice variants do not, by themselves, establish a visible shift.
Compare them with the ~150 kDa apparent band and the listed splice changes before assigning an isoform. The features give no signal peptide, propeptide or glycosylation sites to support those explanations. An unexpected band cannot be identified from migration alone.
Compare them with the ~150 kDa apparent band and the listed splice changes before assigning an isoform. The features give no signal peptide, propeptide or glycosylation sites to support those explanations. An unexpected band cannot be identified from migration alone.
Boster reagents
MYO6 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-MYO6 Antibody Picoband®
Cat # A02627-2
The catalog reports one anti-MYO6 antibody, A02627-2, with stated human and mouse reactivity. Its Western blot image shows an approximately 150 kDa band in human A431 cell and mouse brain lysates. The supplied evidence is limited to this product example.
Which to pick: A02627-2 is the only listed option. Its Western blot image documents human A431 cells and mouse brain under the reported conditions; use those examples when judging fit for your sample.
Source: BosterBio MYO6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UM54.
- Human Protein Atlas. MYO6 tissue expression.
- PMC7196043 — target-verified WB comparison
- PMC11515482 — target-verified WB comparison
- PMC8515727 — target-verified WB comparison