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- Table of Contents
MYOG / Myogenin · Western blot design guide
Design a Western Blot for MYOG
Real validated MYOG Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MYOG 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
MYOG 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 | ~25 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MYOG Western Blot Protocols
The M03238-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | C2C12 cell lysate (catalog M03238-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03238-1; 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 MYOG Western Blot Band Size?
Myogenin is predicted at 25 kDa; phosphorylation and homodimerization could affect appearance, but their effects on migration are unproven.
| Band near 25 kDa | Consistent with the predicted size of myogenin; confirm identity with controls |
| Closely spaced bands near 25 kDa | Could reflect phosphorylation at Ser77, Ser79, or Thr87; distinct migration is unproven |
| Higher band near 50 kDa | Could represent a retained homodimer, but its stability during SDS-PAGE is unestablished |
| Weak band in whole-cell lysate | Nuclear myogenin may be more concentrated in a nuclear extract |
💡Expected MYOG appearanceUniProt predicts myogenin at 25 kDa; no empirical band size or demonstrated migration shift is supplied, so confirm any candidate band with ordinary identity controls.
| Predicted molecular mass | Places the unmodified monomer near 25 kDa |
| Phosphoserine 77 by CaMK2G | May alter migration slightly; no shift is demonstrated |
| Phosphoserine 79 by CaMK2G | May alter migration slightly; no shift is demonstrated |
| Phosphothreonine 87 by CaMK2G | May alter migration slightly; no shift is demonstrated |
| Homodimer formation | Could produce a higher band if the dimer survives sample preparation; this is unproven |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear myogenin may be diluted in whole-cell lysate | Check nuclear extraction and use a positive-control lysate |
| Band higher than expected | Phosphorylation or a retained homodimer is possible, but neither shift is demonstrated | Compare phosphatase-treated and untreated samples and verify band identity |
| Band lower than expected | No listed cleavage feature explains a smaller product | Check sample integrity and verify band identity with a specificity control |
| Multiple bands | Phosphorylated states are possible, but distinct bands are unproven | Compare phosphatase-treated samples and use a specificity control |
| Weak or no signal | Nuclear localization can reduce the target concentration in whole-cell lysate | Test a nuclear extract and a positive-control lysate |
Experiment evidence
Western blot images showing MYOG band
M03238-1 · MYOG
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MYOG Western blot
🧪For positive controls for MYOG in Western blot, you can use no HPA-validated tissue or cell line from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA tissue data are unavailable, so a tissue-based positive or negative control cannot be identified.
HPA tissue expression evidence for MYOG
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced MYOG Western Blot Tips
Deeper troubleshooting and optimisation questions for MYOG, answered from its protein features.
How should MYOG 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 MYOG isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to MYOG isoforms.
Which MYOG phosphorylation sites should I consider?
PTM · The listed sites are Ser77, Ser79, and Thr87, each phosphorylated by CaMK2G. These are UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish a detectable band shift.
Does this guide establish induction of MYOG?
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 MYOG Western blot?
Transfer · MYOG is predicted at 25 kDa. Validate that your transfer setup retains and transfers proteins near that size, using a nearby molecular weight marker. The supplied features do not specify a membrane or transfer conditions.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03238-1 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 I quantify a MYOG band?
Quantitation · MYOG is annotated as nuclear. Compare consistently prepared samples and quantify the band identified near its predicted 25 kDa position. The supplied features give no observed band size, so validate band identity before using its intensity to compare samples.
Should MYOG run at exactly 25 kDa?
Interpretation · MYOG has a predicted mass of 25 kDa, but no observed band size is supplied. Use 25 kDa as a reference, not an exact expected position. The listed phosphorylation sites alone cannot establish a visible shift or explain a mass difference.
MYOG is annotated for myogenesis, differentiation, and transcription regulation. It is recruited to late myogenic gene promoter sequences. Compare samples at defined differentiation stages, but do not assume the record establishes when MYOG abundance rises.
Check band identity before assigning an unexpected band to MYOG. The record lists phosphorylation and interactions with MYOG and E12, but neither feature alone establishes a second Western blot band. No observed band pattern is supplied.
MYOG is annotated for myogenesis, differentiation, and transcription regulation. It is recruited to late myogenic gene promoter sequences. Compare samples at defined differentiation stages, but do not assume the record establishes when MYOG abundance rises.
Check band identity before assigning an unexpected band to MYOG. The record lists phosphorylation and interactions with MYOG and E12, but neither feature alone establishes a second Western blot band. No observed band pattern is supplied.
Boster reagents
MYOG 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-Myogenin Monoclonal Antibody
Cat # M03238-1
The catalog reports one anti-MYOG antibody, M03238-1, for Western blotting. It is catalogued as reactive with human, mouse, and rat, and its WB image shows Myogenin expression in C2C12 cell lysate. The supplied evidence does not document other tested specimens.
Which to pick: M03238-1 is the only listed option and has a WB image using C2C12 cell lysate. Check its reported human, mouse, and rat reactivity against your sample; the image documents the C2C12 context specifically.
Source: BosterBio MYOG gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P15173.
- Human Protein Atlas. MYOG tissue expression.
- PMC7815108 — target-verified WB comparison
- PMC11468980 — target-verified WB comparison
- PMC12176917 — target-verified WB comparison