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- Table of Contents
MYO7A / Unconventional myosin-VIIa · Western blot design guide
Design a Western Blot for MYO7A
Source-linked MYO7A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYO7A 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
MYO7A 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 | ~254.4 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Adrenal gland (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 | 8 isoform(s) |
Section 1
Source-Linked MYO7A Western Blot Protocol Options
The M03915 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Y79 cell lysate (catalog M03915) |
| 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 | M03915; 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 MYO7A Western Blot Band Size?
MYO7A is predicted at 254.4 kDa; isoforms and possible homodimerization could affect migration, but no empirical band or demonstrated shift is supplied.
| Band near 254.4 kDa | Consistent with full-length MYO7A; identity requires confirmation |
| Band near twice the monomer size | Could reflect a homodimer if the proposed association survives electrophoresis |
| Several bands at different positions | Could reflect MYO7A isoforms; distinct migration is unconfirmed |
| Weak or absent lysate band | Cytoskeleton-associated MYO7A may be poorly extracted |
💡Expected MYO7A appearanceMYO7A has a predicted mass of 254.4 kDa, but no empirical band size is supplied; confirm any band near that size with antibody specificity and loss-of-target controls.
| Predicted MYO7A mass | Full-length sequence predicts 254.4 kDa |
| Possible coiled-coil homodimer | Could migrate near twice the monomer size if the association survives electrophoresis |
| Isoforms 1, 2, 3, and 4 | May differ in size; individual masses and migration are unspecified |
| Isoforms 5, 6, 7, and 8 | May differ in size; individual masses and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeleton-associated MYO7A may be poorly extracted | Check insoluble fractions and a validated positive-control lysate |
| Band higher than expected | The proposed homodimer may persist during electrophoresis | Compare denaturing conditions and confirm identity by target depletion |
| Band lower than expected | A shorter isoform is possible, but its mass is unspecified | Check epitope coverage and confirm the band by target depletion |
| Multiple bands | Eight annotated isoforms could contribute, but distinct bands are unproven | Compare isoform information and use target depletion to identify specific bands |
| Weak or no signal | MYO7A may be under-extracted from the cytoskeleton | Check extraction efficiency and a validated positive-control lysate |
| Fragments below expected size | MYO7A may have degraded during sample preparation | Prepare fresh lysate with protease inhibitors and confirm band identity |
Experiment evidence
Western blot images showing MYO7A band
M03915 · MYO7A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYO7A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYO7A in Western blot, you can use adrenal gland lysate, which HPA scores High.
Positive control: Adrenal gland (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: MYO7A is intracellular, but confirm that HPA's not-detected adipose tissue lacks a Western-blot signal.
HPA tissue expression evidence for MYO7A
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | 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 → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYO7A Western Blot Tips
Deeper troubleshooting and optimisation questions for MYO7A, answered from its protein features.
How should MYO7A 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 MYO7A isoforms could produce smaller bands?
Isoforms · UniProt lists eight isoforms. Isoforms 3 and 4 lack canonical residues 1201–2215, a substantial deletion. Isoform 5 lacks residues 284–360 and 519–564; several others have shorter deletions. These sequence differences make distinct band sizes plausible, but do not establish where any isoform migrates.
Check the epitope against the listed alternative sequences. An epitope within canonical residues 1201–2215 will be absent from isoforms 3 and 4; residues 284–360 or 519–564 will be absent from isoform 5. Choose a region retained by the isoforms you intend to measure.
Check the epitope against the listed alternative sequences. An epitope within canonical residues 1201–2215 will be absent from isoforms 3 and 4; residues 284–360 or 519–564 will be absent from isoform 5. Choose a region retained by the isoforms you intend to measure.
Can MYO7A phosphorylation explain a shifted band?
PTM · UniProt lists phosphoserine 1569 and phosphothreonine 1571, using canonical sequence coordinates. Both sites fall within the region absent from isoforms 3 and 4. Their presence in the feature list does not establish a visible shift. If testing phosphorylation, compare matched samples with and without phosphatase treatment.
Does this guide establish induction of MYO7A?
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 MYO7A Western blot?
Transfer · MYO7A has a predicted canonical mass of 254.4 kDa. Choose transfer conditions suitable for a protein of that size, then check the membrane and remaining gel to see whether it transferred. The supplied features do not specify a validated transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03915 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 can I quantify MYO7A across samples?
Quantitation · Use an antibody epitope retained by the isoforms you want to compare, and quantify the same band or defined set of bands in each sample. UniProt lists eight isoforms with sequence differences, so an antibody that misses an isoform would measure only part of the MYO7A signal.
Should MYO7A migrate at its predicted 254.4 kDa?
Interpretation · The canonical sequence has a predicted mass of 254.4 kDa. No observed band position is supplied, so compare the blot with a molecular weight marker without assuming an exact apparent mass. The listed features alone cannot explain a difference between predicted and observed migration.
Compare their sizes and antibody epitopes with the listed isoform deletions, especially residues 1201–2215 missing from isoforms 3 and 4. The canonical predicted mass is 254.4 kDa, but no observed band positions are supplied. A band’s position alone cannot identify its isoform or establish that phosphorylation caused it.
Compare their sizes and antibody epitopes with the listed isoform deletions, especially residues 1201–2215 missing from isoforms 3 and 4. The canonical predicted mass is 254.4 kDa, but no observed band positions are supplied. A band’s position alone cannot identify its isoform or establish that phosphorylation caused it.
Boster reagents
MYO7A 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-MYO7A Monoclonal Antibody
Cat # M03915
The catalog reports M03915, an anti-MYO7A monoclonal antibody with reported Human, Mouse, and Rat reactivity. Its Western blot image shows Myosin VIIa expression in Y79 cell lysate; the supplied evidence documents this sample context only.
Which to pick: M03915 is the only listed option. It has a Western blot image from Y79 cell lysate and reported Human, Mouse, and Rat reactivity. Check suitability for your specific sample and conditions.
Source: BosterBio MYO7A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13402.
- Human Protein Atlas. MYO7A tissue expression.
- PMC5385277 — target-verified WB comparison
- PMC3927955 — target-verified WB comparison
- PMC9996380 — target-verified WB comparison