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- Table of Contents
MYL6 / Myosin light polypeptide 6 · Western blot design guide
Design a Western Blot for MYL6
Source-linked MYL6 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYL6 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
MYL6 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 | ~16.9 kDa | |
| Observed band | ~16 kDa | |
| Gel | 5–20% (catalog A09646-1) | |
| Positive control | Skeletal muscle (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 C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked MYL6 Western Blot Protocol Options
The A09646-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SiHa, human Hela, human THP-1 (catalog A09646-1) |
| Gel % | 5–20% (catalog A09646-1) |
| Load | 30 ug; reducing conditions (catalog A09646-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A09646-1) |
| Membrane | nitrocellulose membrane (catalog A09646-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A09646-1) |
| Primary antibody | A09646-1 · 0.5 μg/mL (catalog A09646-1) |
| Primary incubation | overnight at 4°C (catalog A09646-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A09646-1) |
| Secondary incubation | 1.5 hour at RT (catalog A09646-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A09646-1) |
| Detection | ECL (catalog A09646-1) |
Section 2
What Is the Expected MYL6 Western Blot Band Size?
MYL6 is predicted at 16.9 kDa and observed at ~16 kDa; the cause of the small difference is not established.
| Band at ~16 kDa | Empirical MYL6 band observed in reducing whole-cell lysates |
| Band near 16.9 kDa | Consistent with the UniProt-predicted MYL6 mass; confirm identity with controls |
| Several bands near the monomer region | The Non-muscle and Smooth muscle isoforms are possible contributors, but distinct migration is unverified |
| One band despite two named isoforms | The isoforms need not resolve as separate bands |
💡Expected MYL6 appearanceMYL6 has a predicted mass of 16.9 kDa and an empirical band at ~16 kDa in reducing whole-cell lysates; use ordinary band-identity controls to verify the signal.
| UniProt-predicted MYL6 mass | 16.9 kDa predicted; the empirical band is ~16 kDa |
| Alternative splicing | Produces named isoforms, but their migration difference is not established |
| Non-muscle isoform | Its individual mass and band position are not supplied |
| Smooth muscle isoform | Its individual mass and band position are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MYL6 signal may be below detection in the tested sample | Check loading and detection against a validated positive whole-cell lysate |
| Band higher than expected | The band's identity or migration is uncertain; no glycosylation or disulfide is listed | Compare with a positive lysate and verify antibody specificity |
| Band lower than expected | A different isoform or an unrelated band is possible; isoform masses are unknown | Check band identity with an independent antibody or MYL6 depletion |
| Multiple bands | Named isoforms are possible contributors, but their separation is unverified | Compare samples and verify each band with MYL6 depletion |
| Weak or no signal | Insufficient MYL6 or assay sensitivity | Check protein loading, transfer and a validated positive lysate |
| Fragments below expected size | Sample degradation or unrelated antibody binding | Use fresh protease-protected lysate and verify band identity |
Experiment evidence
Western blot images showing MYL6 band
A09646-1 · MYL6
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYL6 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYL6 in Western blot, you can use skeletal muscle lysate, which HPA rates as highly positive.
Positive control: Skeletal muscle (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 adipose tissue as not detected, so a negative tissue control is feasible.
HPA tissue expression evidence for MYL6
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 |
|---|---|---|---|---|
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYL6 Western Blot Tips
Deeper troubleshooting and optimisation questions for MYL6, answered from its protein features.
How should MYL6 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.
Can the two MYL6 isoforms be distinguished by band position?
Isoforms · UniProt lists Non-muscle and Smooth muscle isoforms. In the Smooth muscle isoform, residues 143–151 change from AFVRHILSG to ELVRMVLNG. This is a nine-residue substitution, so separate bands are not established by the sequence feature alone. Check whether the antibody recognizes both sequences.
If the antibody recognizes both isoforms and they overlap near 16 kDa, the band may represent their combined signal. The listed isoform difference is at residues 143–151; check antibody recognition of that region before treating the signal as specific to one isoform.
If the antibody recognizes both isoforms and they overlap near 16 kDa, the band may represent their combined signal. The listed isoform difference is at residues 143–151; check antibody recognition of that region before treating the signal as specific to one isoform.
Which MYL6 modifications should I consider when interpreting bands?
PTM · The supplied UniProt coordinates list N-acetylcysteine at position 2, phosphoserine at 57, and the annotation ‘N6-acetyllysine Phosphoserine’ at 81. Keep those coordinates and the position-81 wording when comparing antibody information; numbering conventions may differ. These annotations alone do not establish a visible band shift.
Does this guide establish induction of MYL6?
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 MYL6 Western blot?
Transfer · MYL6 has a predicted mass of 16.9 kDa and an observed band near 16 kDa. Check that your chosen transfer conditions retain a band in that range on the membrane. The supplied features do not specify a preferred transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09646-1 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 MYL6 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.
Why is MYL6 observed near 16 kDa instead of 16.9 kDa?
Interpretation · The observed band is approximately 16 kDa, close to the predicted 16.9 kDa. The supplied features do not establish what causes the difference. Use the observed position as an approximate guide, not an exact mass check.
Start with the expected band near 16 kDa. MYL6 has two listed isoforms and belongs to a myosin hexamer containing two heavy and four light chains, but those features alone do not identify an extra band. Verify antibody recognition and compare the unexpected band with the expected MYL6 signal before assigning it to an isoform, modification, or complex.
Start with the expected band near 16 kDa. MYL6 has two listed isoforms and belongs to a myosin hexamer containing two heavy and four light chains, but those features alone do not identify an extra band. Verify antibody recognition and compare the unexpected band with the expected MYL6 signal before assigning it to an isoform, modification, or complex.
Boster reagents
MYL6 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-MYL6 Antibody Picoband®
Cat # A09646-1
The catalog reports one anti-MYL6 antibody for Western blotting. Its product image shows an approximately 16 kDa band in human SiHa, HeLa, and THP-1 whole cell lysates using 0.5 μg/mL primary antibody. No independent validation is supplied.
Which to pick: A09646-1 is the only listed option. It reports human reactivity and includes a Western blot image from three human cell lysates; those samples are the documented test contexts.
Source: BosterBio MYL6 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P60660.
- Human Protein Atlas. MYL6 tissue expression.
- PMC8869929 — target-verified WB comparison
- PMC11430384 — target-verified WB comparison
- PMC8635237 — target-verified WB comparison