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- Table of Contents
MYL7 / Myosin regulatory light chain 2, atrial isoform · Western blot design guide
Design a Western Blot for MYL7
Real validated MYL7 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYL7 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
MYL7 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 | ~19.4 kDa | |
| Observed band | Approximately 19 kDa | |
| Gel | 12% (catalog A11956-1) | |
| Positive control | Heart muscle (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MYL7 Western Blot Protocols
The A11956-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat heart, mouse heart (catalog A11956-1) |
| Gel % | 12% (catalog A11956-1) |
| Load | 30 ug; reducing conditions (catalog A11956-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A11956-1) |
| Membrane | nitrocellulose membrane (catalog A11956-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A11956-1) |
| Primary antibody | A11956-1 · 1:1000 (catalog A11956-1) |
| Primary incubation | overnight at 4°C (catalog A11956-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A11956-1) |
| Secondary incubation | 1.5 hour at RT (catalog A11956-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A11956-1) |
| Detection | ECL (catalog A11956-1) |
Section 2
What Is the Expected MYL7 Western Blot Band Size?
MYL7 is predicted at 19.4 kDa and observed at approximately 19 kDa; the small difference has no established cause in the supplied evidence.
| Band at approximately 19 kDa | Empirical MYL7 band, consistent with the 19.4 kDa predicted mass |
| Single sharp band near 19 kDa | Consistent with the one listed MYL7 isoform |
| Slightly shifted band near 19 kDa | Could reflect phosphorylation at Ser22 or Ser23; a visible shift is unproven |
| Closely spaced bands near 19 kDa | Could reflect different phosphorylation states; their separation is unproven |
💡Expected MYL7 appearanceMYL7 has a predicted mass of 19.4 kDa and an observed band at approximately 19 kDa in reducing heart lysates; confirm identity with appropriate antibody and sample controls.
| Predicted MYL7 mass | 19.4 kDa predicted; the observed band is approximately 19 kDa |
| N-acetylalanine at residue 2 | Adds a small modification; a visible migration change is not established |
| Phosphoserine at residue 22 | May affect migration; no distinct band size is established |
| Phosphoserine at residue 23 | May affect migration; no distinct band size is established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | Identity or phosphorylation-dependent migration is uncertain | Compare with heart lysate and check antibody specificity; assess phosphatase sensitivity |
| Band lower than expected | Possible sample degradation; no cleavage feature is listed | Use fresh lysate with protease inhibitors and confirm band identity |
| Multiple bands | Phosphorylation states are possible, but distinct bands are unproven | Compare phosphatase-treated samples and confirm with a second antibody |
| Weak or no signal | Insufficient detectable MYL7 or loss during preparation or transfer | Check loading and transfer using heart lysate as a positive control |
| Fragments below expected size | Possible proteolysis during sample preparation | Prepare fresh lysate with protease inhibitors and verify with a second antibody |
Experiment evidence
Western blot images showing MYL7 band
A11956-1 · MYL7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYL7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MYL7 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 the samples.
⚠️Feasibility: HPA reports high expression in heart muscle and no detection in adipose tissue, making tissue controls feasible.
HPA tissue expression evidence for MYL7
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 → |
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 MYL7 Western Blot Tips
Deeper troubleshooting and optimisation questions for MYL7, answered from its protein features.
How should MYL7 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 MYL7 isoforms explain additional bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning additional bands to MYL7 isoforms.
Which MYL7 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylalanine at position 2 and phosphoserine at positions 22 and 23. These features identify possible modified forms, but they do not establish that separate bands will be visible. The positions use the supplied UniProt sequence numbering.
Phosphoserine is listed at UniProt positions 22 and 23. A phospho-specific signal could reflect phosphorylation at the site recognized by the antibody; check its stated site and numbering against these UniProt coordinates. The features alone do not show when phosphorylation occurs or how much signal to expect.
Phosphoserine is listed at UniProt positions 22 and 23. A phospho-specific signal could reflect phosphorylation at the site recognized by the antibody; check its stated site and numbering against these UniProt coordinates. The features alone do not show when phosphorylation occurs or how much signal to expect.
Does this guide establish induction of MYL7?
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.
How should transfer be checked for MYL7?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A11956-1 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 MYL7 bands be quantified?
Quantitation · Define whether the measurement is total MYL7 or phosphorylation at UniProt Ser22 or Ser23. Quantify the corresponding signal consistently across samples; the listed phosphorylation sites do not establish that a shifted band represents all phosphorylated MYL7.
How does the observed MYL7 band compare with its predicted mass?
Interpretation · The approximately 19 kDa observed band is close to MYL7’s predicted 19.4 kDa mass. Its listed modifications do not establish a visible shift or explain any small difference between apparent and calculated mass.
The record lists a 175-residue protein with one isoform, no signal peptide or propeptide, and no glycosylation sites. It also describes myosin as a hexamer of two heavy chains and four light chains. These features alone cannot identify an unexpected band; confirm its identity before assigning it to a modification or myosin-associated species.
The record lists a 175-residue protein with one isoform, no signal peptide or propeptide, and no glycosylation sites. It also describes myosin as a hexamer of two heavy chains and four light chains. These features alone cannot identify an unexpected band; confirm its identity before assigning it to a modification or myosin-associated species.
Boster reagents
MYL7 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-MYL7 Antibody Picoband®
Cat # A11956-1
The catalog reports A11956-1, a rabbit polyclonal anti-MYL7 antibody, for Western blotting with reported human, mouse, and rat reactivity. Its WB image shows a band near the expected 19 kDa in rat and mouse heart lysates; the supplied image does not document human samples.
Which to pick: A11956-1 is the only listed option. Its WB example uses rat and mouse heart lysates under reducing conditions at a 1:1000 primary dilution. For human samples, reactivity is listed, but no human WB example is supplied.
Source: BosterBio MYL7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q01449.
- Human Protein Atlas. MYL7 tissue expression.
- PMC12890996 — target-verified WB comparison
- PMC6089907 — target-verified WB comparison
- PMC3407188 — target-verified WB comparison