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- Table of Contents
PAX7 / Paired box protein Pax-7 · Western blot design guide
Design a Western Blot for PAX7
Real validated PAX7 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PAX7 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
PAX7 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 | ~55.1 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 | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated PAX7 Western Blot Protocols
The M00845-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa lysate (catalog M00845-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M00845-1; 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 PAX7 Western Blot Band Size?
PAX7 is predicted at 55.1 kDa; isoforms and acetylation are documented, but their effects on band migration are not demonstrated.
| Band near 55.1 kDa | Consistent with the predicted PAX7 mass; confirm identity with antibody controls |
| Several bands | Could reflect PAX7 isoforms 1, 2, and 3 if their migration differs; identity requires confirmation |
| Little or no band in a cytosolic fraction | Consistent with PAX7 localization to the nucleus |
| No clear shift between samples | Acetylation at Lys105 or Lys139 does not establish a resolvable size change |
💡Expected PAX7 appearancePAX7 has a predicted mass of 55.1 kDa, but no empirical band size is supplied; use band identity controls because isoform migration and effects of acetylation are not established.
| UniProt predicted mass | Places the reference band near 55.1 kDa |
| PAX7 isoform 1 | May differ in size from other isoforms; its mass and migration are unspecified |
| PAX7 isoform 2 | May differ in size from other isoforms; its mass and migration are unspecified |
| PAX7 isoform 3 | May differ in size from other isoforms; its mass and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear PAX7 may be poorly recovered in the lysate | Check nuclear extraction and include a positive control |
| Band higher than expected | The supplied features do not establish a higher migrating PAX7 species | Compare with a positive control and verify antibody specificity |
| Band lower than expected | An isoform could differ in size, but isoform masses are unavailable | Check isoform expression and verify band identity |
| Multiple bands | PAX7 has three named isoforms, but their band positions are unknown | Check isoform expression and use a PAX7 depletion control |
| Weak or no signal | Nuclear localization may limit PAX7 recovery | Test a nuclear fraction alongside a positive control |
Experiment evidence
Western blot images showing PAX7 band
M00845-1 · PAX7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PAX7 Western blot
🧪For positive controls for PAX7 in Western blot, you can use no HPA-supported tissue or cell sample 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 provides no expression data, so suitable tissue controls cannot be confirmed.
HPA tissue expression evidence for PAX7
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 PAX7 Western Blot Tips
Deeper troubleshooting and optimisation questions for PAX7, answered from its protein features.
How should PAX7 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 PAX7 isoforms produce different bands?
Isoforms · Three isoforms are listed. Isoform 2 lacks residues 151–152, while isoforms 1 and 2 replace the canonical sequence at 468–505 with a different sequence. These changes can alter molecular mass, but the features do not establish how far the bands separate on a blot.
How should PAX7 acetylation be considered when interpreting bands?
PTM · UniProt lists N6-acetyllysine at positions 105 and 139 in its sequence numbering. Record that numbering when comparing antibody or paper annotations. These modifications alone do not demonstrate a resolvable band shift.
Does this guide establish induction of PAX7?
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 PAX7?
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 M00845-1 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 PAX7 bands be quantified across samples?
Quantitation · Use a consistently defined PAX7 band or set of bands across samples. Because three isoforms are listed, state whether the measurement covers one band or sums multiple bands; do not assume every band represents the same isoform.
Should PAX7 migrate at its predicted 55.1 kDa?
Interpretation · 55.1 kDa is the predicted mass; no observed band position is supplied. Compare candidate bands with this estimate, but do not assign a band solely by its apparent mass. The listed acetylation sites and isoforms do not establish a visible shift.
Check the antibody epitope against the isoform sequences. Isoforms 1 and 2 have a replacement at canonical positions 468–505, and isoform 2 also lacks residues 151–152. An epitope overlapping either changed region may affect which isoforms the antibody detects.
PAX7 is annotated as nuclear. Include the nuclear fraction when preparing samples, and keep the fraction or preparation method consistent across lanes used for comparison.
Compare their positions with the 55.1 kDa predicted mass and consider the listed isoform sequence changes. The supplied features do not provide an observed band position or establish the identity of an extra band. Check whether the antibody epitope is shared across the isoforms before assigning it.
Check the antibody epitope against the isoform sequences. Isoforms 1 and 2 have a replacement at canonical positions 468–505, and isoform 2 also lacks residues 151–152. An epitope overlapping either changed region may affect which isoforms the antibody detects.
PAX7 is annotated as nuclear. Include the nuclear fraction when preparing samples, and keep the fraction or preparation method consistent across lanes used for comparison.
Compare their positions with the 55.1 kDa predicted mass and consider the listed isoform sequence changes. The supplied features do not provide an observed band position or establish the identity of an extra band. Check whether the antibody epitope is shared across the isoforms before assigning it.
Boster reagents
PAX7 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-PAX7 Rabbit Monoclonal Antibody
Cat # M00845-1
The catalog reports one anti-PAX7 antibody for Western blot: M00845-1, a rabbit monoclonal with reported human, mouse, and rat reactivity. Its WB image shows PAX7 expression in HeLa lysate; the supplied evidence does not show mouse or rat samples.
Which to pick: M00845-1 is the only listed option. Its WB image uses HeLa lysate, making that the documented test context. Human, mouse, and rat reactivity is reported, but the supplied image supports only the HeLa example.
Source: BosterBio PAX7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P23759.
- Human Protein Atlas. PAX7 tissue expression.
- PMC12079090 — target-verified WB comparison
- PMC9149870 — target-verified WB comparison
- PMC5646146 — target-verified WB comparison