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- Table of Contents
MSX1 / Homeobox protein MSX-1 · Western blot design guide
Design a Western Blot for MSX1
Real validated MSX1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MSX1 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
MSX1 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 | ~31.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cervix (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 | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MSX1 Western Blot Protocols
The M01701 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | M01701; 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 MSX1 Western Blot Band Size?
MSX1 is predicted at 31.5 kDa; no empirical band is supplied, and the listed features do not demonstrate altered migration.
| Single band near 31.5 kDa | Consistent with predicted MSX1 size; confirm identity with controls |
| Band enriched in nuclear extract | Consistent with MSX1 nuclear localization |
| Weak band in cytoplasmic extract | Consistent with MSX1 nuclear localization |
| Several bands at different sizes | Band identities need confirmation; no distinct isoforms are listed |
💡Expected MSX1 appearanceUniProt predicts MSX1 at 31.5 kDa, but no empirical band size is supplied; confirm a candidate band with appropriate identity controls.
| UniProt predicted mass | Sets a reference size of 31.5 kDa |
| Predicted molecular weight in daltons | Equals 31,496 Da, approximately 31.5 kDa |
| Predicted polypeptide size | Places a candidate full-length band near 31.5 kDa |
| UniProt mass reference | Provides a size estimate rather than a measured migration position |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear MSX1 may be poorly represented in the preparation | Check a nuclear extract and a positive control |
| Band higher than expected | Band identity or migration is unestablished | Compare with an MSX1 depletion control |
| Band lower than expected | Band identity or integrity is unestablished | Check sample handling and use an MSX1 depletion control |
| Multiple bands | Only one MSX1 isoform is listed | Identify the specific band with an MSX1 depletion control |
| Weak or no signal | The preparation may contain little nuclear MSX1 | Check nuclear enrichment and include a positive control |
| Fragments below expected size | Sample degradation is possible | Prepare fresh lysate with protease inhibitors and confirm band identity |
Experiment evidence
Western blot images showing MSX1 band
M01701 · MSX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MSX1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MSX1 in Western blot, you can use cervix tissue, where HPA reports high expression.
Positive control: Cervix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: MSX1 is nuclear, so nuclear enrichment may help if whole-tissue lysate gives a weak signal.
HPA tissue expression evidence for MSX1
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 |
|---|---|---|---|---|
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | ciliated epithelial 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 → |
| 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 MSX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MSX1, answered from its protein features.
Where should the main MSX1 band appear?
Band shift · The 303-residue canonical MSX1 sequence has a predicted mass of 31.5 kDa. Use this as a reference, not a guaranteed apparent position; no observed band size is supplied.
Could an MSX1 isoform explain an extra band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning a second band to an annotated MSX1 isoform.
Could modification change the apparent MSX1 band?
PTM · The record includes Ubl conjugation and isopeptide bond keywords, but lists no modified residue or coordinate. These features alone do not establish a visible band shift or explain a difference from 31.5 kDa.
Does this guide establish induction of MSX1?
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 MSX1?
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 M01701 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 MSX1 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Which sample fraction is most relevant for MSX1 detection?
Interpretation · MSX1 is annotated as nuclear. Check the nuclear fraction when assessing detection; its interaction with PIAS1 is reported to be required for localization to the nuclear periphery by similarity.
Use comparable nuclear sample preparations when comparing MSX1 signal. Its nuclear localization, including PIAS1-associated localization to the nuclear periphery, makes differences in fraction recovery relevant to interpretation.
Compare them with the 31.5 kDa predicted mass, but do not assign their identity from size alone. The supplied record has no observed band size or annotated alternative sequence, and its Ubl conjugation keyword does not establish a visible shifted band.
Use comparable nuclear sample preparations when comparing MSX1 signal. Its nuclear localization, including PIAS1-associated localization to the nuclear periphery, makes differences in fraction recovery relevant to interpretation.
Compare them with the 31.5 kDa predicted mass, but do not assign their identity from size alone. The supplied record has no observed band size or annotated alternative sequence, and its Ubl conjugation keyword does not establish a visible shifted band.
Boster reagents
MSX1 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-Msx-1 Monoclonal Antibody
Cat # M01701
M01701 is a human-reactive anti-MSX1 monoclonal antibody with a Western blot image using NTERA-2 cell lysate. The supplied caption describes SDS-PAGE but provides incomplete experimental conditions; this image documents one tested sample context.
Which to pick: M01701 is the only listed antibody. Its NTERA-2 lysate Western blot image provides a starting point for human MSX1 experiments; check performance in your own sample and conditions.
Source: BosterBio MSX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P28360.
- Human Protein Atlas. MSX1 tissue expression.
- PMC6091477 — target-verified WB comparison
- PMC9960665 — target-verified WB comparison
- PMC11754605 — target-verified WB comparison