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- Table of Contents
MYH1 / Myosin-1 · Western blot design guide
Design a Western Blot for MYH1
Real validated MYH1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYH1 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
MYH1 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 | ~223.1 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Heart muscle (IHC candidate; verify WB) +1 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MYH1 Western Blot Protocols
The A09420 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A09420) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A09420) |
| Primary antibody | A09420 · 1:410 (catalog A09420) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A09420) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A09420) |
Section 2
What Is the Expected MYH1 Western Blot Band Size?
MYH1 is predicted at 223.1 kDa; its assembly and documented modifications could affect samples, but altered Western-blot migration has not been demonstrated here.
| Band near 223.1 kDa | Consistent with full-length MYH1; confirm identity with controls. |
| Higher molecular weight band | Could reflect incomplete dissociation of a myosin complex; identity requires confirmation. |
| Weak or absent band in a myofibril-poor lysate | MYH1 is localized to myofibrillar thick filaments. |
| Multiple bands near the expected position | MYH1 has documented modification sites, but distinct migrating forms are not established. |
💡Expected MYH1 appearanceFull-length MYH1 has a predicted mass of 223.1 kDa; no empirical Western-blot band size is supplied, so confirm any candidate band with appropriate identity controls.
| Predicted full-length mass | Sets a reference of 223.1 kDa, not a measured migration position. |
| Phosphorylation sites including Thr64 and Thr69 | Modification is documented, but a visible size shift is not established. |
| Trimethyllysine at residue 130 | Modification is documented, but a visible size shift is not established. |
| Pros-methylhistidine at residue 757 | Modification is documented, but a visible size shift is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The sample may contain little myofibrillar MYH1. | Check muscle content and analyze a myofibril-enriched fraction. |
| Band higher than expected | A myosin complex may have dissociated incompletely. | Check sample denaturation and compare with a well-denatured preparation. |
| Band lower than expected | The band may be a fragment or an unrelated protein. | Check sample integrity and confirm identity with an independent antibody. |
| Multiple bands | Fragments or antibody cross-reactivity may contribute; distinct MYH1 isoform bands are not documented here. | Compare antibody epitopes and use a positive-control muscle sample. |
| Weak or no signal | Poor recovery of MYH1 from thick filaments may reduce signal. | Check extraction and loading with a muscle positive control. |
| Fragments below expected size | MYH1 may have degraded during sample preparation. | Process samples promptly with protease inhibitors and compare fresh lysate. |
Experiment evidence
Western blot images showing MYH1 band
A09420 · MYH1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYH1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYH1 in Western blot, you can use heart muscle lysate.
Positive control: Heart 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 MYH1.
⚠️Feasibility: As a myofibrillar protein, MYH1 is best detected in muscle-rich lysate.
HPA tissue expression evidence for MYH1
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 |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | 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 → |
| 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYH1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MYH1, answered from its protein features.
Where should the MYH1 band appear?
Band shift · The predicted mass is 223.1 kDa; use that as a reference when identifying the full-length heavy chain. No observed band position is supplied. The listed modifications alone do not establish a visible shift or explain any difference between apparent and predicted mass.
Does UniProt support multiple MYH1 isoform bands?
Isoforms · This record lists one isoform and no alternative sequence. It therefore provides no sequence-based explanation for multiple MYH1 bands. Confirm band identity before labeling additional bands as isoforms.
Could phosphorylation affect MYH1 band interpretation?
PTM · UniProt lists phosphosites including Thr64, Thr69, Ser1092, Ser1096, Ser1730, and Ser1739. These are UniProt coordinates; antibody or paper numbering may differ. A changed band pattern cannot be assigned to phosphorylation from these features alone. Compare matched samples and use a site-specific assay if phosphorylation is the question.
Does this guide establish induction of MYH1?
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 MYH1 Western blot?
Transfer · MYH1 is a 223.1 kDa heavy chain, so check that your chosen transfer conditions move proteins near this mass onto the membrane. Verify transfer in that region with a total-protein stain and adjust the conditions if the high-mass region transfers poorly.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09420 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 MYH1 signal be quantified?
Quantitation · MYH1 is located in myofibrillar thick filaments. Compare samples prepared with a consistent myofibril extraction procedure, quantify the full-length band, and normalize to an appropriate loading measure from the same samples. The feature record provides no empirical band intensity or expected change between conditions.
Are methylated MYH1 residues relevant to antibody selection?
Interpretation · UniProt lists trimethyllysine at position 130 and Pros-methylhistidine at position 757, using UniProt coordinates. If an antibody targets either region, check whether its validation covers the relevant modified residue. These sites do not, by themselves, predict a resolvable band shift.
Start with the predicted 223.1 kDa mass and confirm which band represents intact MYH1. The record describes a myofibrillar heavy chain in a complex with light chains and lists phosphorylation and methylation sites. Those features alone cannot identify an unexpected band or prove a modification-dependent shift.
Start with the predicted 223.1 kDa mass and confirm which band represents intact MYH1. The record describes a myofibrillar heavy chain in a complex with light chains and lists phosphorylation and methylation sites. Those features alone cannot identify an unexpected band or prove a modification-dependent shift.
Boster reagents
MYH1 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-Myosin-1 MYH1 Antibody
Cat # A09420
A09420 is an anti-MYH1 antibody listed as reactive with human, mouse, and rat. Its Western blot image uses extracts from various cell lines at 1:410 dilution and 25 µg per lane; the caption does not identify the cell lines or species.
Which to pick: A09420 is the only listed option and has a Western blot image. Check that its listed reactivity covers your species, and use the reported 1:410 dilution as a starting point.
Source: BosterBio MYH1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P12882.
- Human Protein Atlas. MYH1 tissue expression.
- PMC6814601 — target-verified WB comparison
- PMC11425116 — target-verified WB comparison
- PMC13242725 — target-verified WB comparison