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- Table of Contents
MYH8 / Myosin-8 · Western blot design guide
Design a Western Blot for MYH8
Source-linked MYH8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYH8 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
MYH8 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 | ~222.8 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Skeletal 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
Source-Linked MYH8 Western Blot Protocol Options
The A08502 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog A08502) |
| 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 | A08502 · (A) 0.5 and (B) 1 μg/ml (catalog A08502) |
| 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 MYH8 Western Blot Band Size?
MYH8 has a predicted heavy-chain mass of 222.8 kDa; its listed modifications and myosin assembly could affect appearance, but altered migration is not demonstrated.
| Band near 222.8 kDa | consistent with the predicted MYH8 heavy chain; confirm identity with controls |
| Higher molecular mass band | may reflect incomplete dissociation of the myosin complex |
| Nearby doublet | different phosphorylation states are possible, but distinct migration is unproven |
| Faint or absent band in cell lysate | MYH8 is a myofibrillar muscle protein and may be scarce in the sampled cells |
💡Expected MYH8 appearanceThe predicted MYH8 heavy chain is 222.8 kDa; no empirical band size is supplied, and band identity needs controls because its listed modifications have no demonstrated migration effect.
| Predicted MYH8 heavy-chain mass | places the unmodified sequence near 222.8 kDa |
| Phosphothreonine at residues 66 and 71 | may affect mobility, but no visible shift is established |
| Phosphotyrosine at residue 389 | may affect mobility, but no visible shift is established |
| Trimethyllysine at residue 132 | adds a small modification; no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MYH8 is localized to muscle myofibrils and may be scarce in the sample | check a skeletal-muscle positive control and retain myofibrillar material during extraction |
| Band higher than expected | the myosin complex may be incompletely dissociated | check denaturation and compare with a well-denatured control |
| Band lower than expected | band identity or protein integrity is uncertain | compare with a positive control and verify specificity with an independent MYH8 antibody |
| Multiple bands | modification states are possible, but distinct MYH8 bands are unproven | compare with a positive control and verify each band with an independent antibody |
| Weak or no signal | the sample may contain little myofibrillar MYH8 | check a skeletal-muscle positive control and confirm protein recovery |
| Fragments below expected size | MYH8 may have degraded during sample preparation | prepare fresh samples with protease inhibitors and compare with a positive control |
Experiment evidence
Western blot images showing MYH8 band
A08502 · MYH8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYH8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYH8 in Western blot, you can use skeletal muscle lysate, the highest-scoring HPA positive tissue.
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: MYH8 is myofibrillar, so skeletal muscle lysate is a suitable positive sample.
HPA tissue expression evidence for MYH8
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 → |
| Testis | Leydig 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 → |
| 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 MYH8 Western Blot Tips
Deeper troubleshooting and optimisation questions for MYH8, answered from its protein features.
How should MYH8 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 MYH8 isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. These features do not support assigning multiple bands to MYH8 isoforms; confirm additional bands with antibody specificity controls.
Which MYH8 modifications matter when interpreting bands?
PTM · UniProt lists 35 modified residues, including phosphorylation at Thr66, Thr71, Tyr389, Ser392, and many later sites. It also lists trimethyllysine at Lys132 and pros-methylhistidine at His756. These are UniProt sequence coordinates; paper or antibody numbering may differ. Their presence does not establish a visible band shift.
The supplied features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for attributing a band difference to glycosylation or cleavage of those peptides.
The supplied features list no glycosylation sites, signal peptide, or propeptide. They therefore provide no basis for attributing a band difference to glycosylation or cleavage of those peptides.
Does this guide establish induction of MYH8?
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 MYH8 Western blot?
Transfer · MYH8 is a 222.8 kDa heavy chain. Choose transfer conditions suitable for a protein of this size, then check transfer efficiency near 223 kDa with a molecular weight marker and inspect the gel for protein left behind.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A08502 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 I quantify MYH8 signal?
Quantitation · MYH8 is located in myofibril thick filaments. Keep sample preparation and loading consistent, verify that the detected band is within the assay’s linear range, and normalize to a suitable loading measure. Compare the same verified band across samples.
What size band should I expect for MYH8?
Interpretation · The predicted mass is 222.8 kDa. No observed band size is supplied, so use this as a reference, not an exact migration position. The listed modifications alone do not establish a visible shift or explain any difference from 222.8 kDa.
Start with the 222.8 kDa prediction and verify antibody specificity and transfer. MYH8 belongs to a myosin complex containing heavy and light chains, but the supplied features do not identify additional MYH8 bands or explain their apparent masses. Do not assign an unexpected band to phosphorylation solely because modified sites are listed.
Start with the 222.8 kDa prediction and verify antibody specificity and transfer. MYH8 belongs to a myosin complex containing heavy and light chains, but the supplied features do not identify additional MYH8 bands or explain their apparent masses. Do not assign an unexpected band to phosphorylation solely because modified sites are listed.
Boster reagents
MYH8 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-8 MYH8 Antibody
Cat # A08502
the supplier A08502 is an anti-MYH8 antibody listed as reactive with human, mouse, and rat. Its Western blot image shows HeLa cell lysate tested at 0.5 and 1 μg/ml. The supplied evidence does not show Western blot results in mouse or rat samples.
Which to pick: A08502 is the only listed MYH8 antibody and has a Western blot image using HeLa cell lysate. For mouse or rat samples, the catalog lists reactivity, but no Western blot example is supplied.
Source: BosterBio MYH8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P13535.
- Human Protein Atlas. MYH8 tissue expression.
- PMC9738036 — target-verified WB comparison
- PMC6284384 — target-verified WB comparison
- PMC6845233 — target-verified WB comparison