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- Table of Contents
MSX2 / Homeobox protein MSX-2 · Western blot design guide
Design a Western Blot for MSX2
Real validated MSX2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MSX2 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
MSX2 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 | ~28.9 kDa | |
| Observed band | ~37 kDa | |
| Gel | 5–20% (catalog A01783-2) | |
| Positive control | Breast (IHC candidate; verify WB) +4 more | |
| Negative control | Appendix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Band above predicted | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MSX2 Western Blot Protocols
The A01783-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human RT4, human MCF-7, human T-47D (catalog A01783-2) |
| Gel % | 5–20% (catalog A01783-2) |
| Load | 30 ug; reducing conditions (catalog A01783-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01783-2) |
| Membrane | nitrocellulose membrane (catalog A01783-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01783-2) |
| Primary antibody | A01783-2 · 0.5 μg/mL (catalog A01783-2) |
| Primary incubation | overnight at 4°C (catalog A01783-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01783-2) |
| Secondary incubation | 1.5 hour at RT (catalog A01783-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01783-2) |
| Detection | ECL (catalog A01783-2) |
Section 2
What Is the Expected MSX2 Western Blot Band Size?
MSX2 is predicted at 28.9 kDa, but antibody QC reports approximately 37 kDa; the cause of this difference is not established.
| Band at approximately 37 kDa in whole-cell lysate | Matches the reported MSX2 antibody QC band; confirm identity with controls |
| Band near 29 kDa | Near the predicted MSX2 mass; identity requires confirmation |
| Band enriched in a nuclear fraction | Consistent with MSX2 nuclear localization |
| Weak band in a cytoplasmic fraction | Consistent with MSX2 nuclear localization |
💡Expected MSX2 appearanceMSX2 has a predicted mass of 28.9 kDa, while antibody QC detected a band at approximately 37 kDa in whole-cell lysate; the cause of this difference is unestablished, so confirm band identity with controls.
| Predicted molecular weight of 28.9 kDa | Sets the calculated size of the MSX2 chain |
| Sequence mass of 28,897 Da | Corresponds to approximately 29 kDa before apparent migration is considered |
| 267-residue sequence | Underlies the calculated 28.9 kDa mass |
| Predicted mass relative to the observed band | Is lower than the reported approximately 37 kDa migration; the reason is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear MSX2 may be poorly recovered | Check nuclear extraction and a nuclear fraction |
| Band higher than expected | The reported MSX2 band migrates at approximately 37 kDa despite its 28.9 kDa predicted mass; the cause is unestablished | Compare with the QC band and verify identity with an independent antibody or MSX2 depletion |
| Band lower than expected | Identity of a lower band is uncertain | Check its response to MSX2 depletion |
| Multiple bands | Additional bands have no established explanation in the supplied MSX2 features | Identify the MSX2-dependent band by depletion or an independent antibody |
| Weak or no signal | Nuclear MSX2 may be underrepresented in the sample | Check a nuclear fraction and sample loading |
| Fragments below expected size | Fragment identity is unestablished | Check sample integrity and whether the bands disappear with MSX2 depletion |
Experiment evidence
Western blot images showing MSX2 band
A01783-2 · MSX2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MSX2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MSX2 in Western blot, you can use breast tissue, which has medium HPA expression.
Positive control: Breast (IHC candidate; verify WB)
Negative control: Appendix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: MSX2 is nuclear, so nuclear enrichment may help if whole-cell signal is weak.
HPA tissue expression evidence for MSX2
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 |
|---|---|---|---|---|
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial 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 → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MSX2 Western Blot Tips
Deeper troubleshooting and optimisation questions for MSX2, answered from its protein features.
How should MSX2 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 an MSX2 isoform explain an additional band?
Isoforms · Only one isoform is listed, with no alternative sequence supplied. These features do not support assigning an additional band to a known MSX2 isoform.
Is a modification known to cause an MSX2 band shift?
PTM · No modified residues or glycosylation sites are listed. The supplied features therefore identify no modification that explains a shift from 28.9 to about 37 kDa.
Does this guide establish induction of MSX2?
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 MSX2?
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 A01783-2 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 the reported 37 kDa band be quantified?
Quantitation · Use the same consistently identified band across samples. The reported apparent mass is about 37 kDa, but the mass difference alone does not establish band identity. Check that the signal is attributable to MSX2 before comparing its intensity.
Why might MSX2 appear near 37 kDa instead of 28.9 kDa?
Interpretation · The reported apparent band is about 37 kDa, while the predicted mass is 28.9 kDa. The supplied features do not establish the cause of this difference. Compare the band with a molecular weight marker and confirm its identity before assigning it to MSX2.
MSX2 is listed as nuclear. A nuclear fraction is therefore relevant when checking whether a detected band follows the expected localization.
MSX2 interacts with MINT by similarity and with XRCC6 (Ku70) and XRCC5 (Ku80). Those interactions alone do not establish that a partner or complex causes an unexpected band. Confirm the band's identity before interpreting it as MSX2.
MSX2 is listed as nuclear. A nuclear fraction is therefore relevant when checking whether a detected band follows the expected localization.
MSX2 interacts with MINT by similarity and with XRCC6 (Ku70) and XRCC5 (Ku80). Those interactions alone do not establish that a partner or complex causes an unexpected band. Confirm the band's identity before interpreting it as MSX2.
Boster reagents
MSX2 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-Hox8/MSX2 Antibody Picoband®
Cat # A01783-2
A01783-2 is a human-reactive anti-MSX2 antibody with a Western blot image from RT4, MCF-7, and T-47D whole-cell lysates. The reported band is approximately 37 kDa, versus an expected 29 kDa; the supplied evidence does not explain this difference.
Which to pick: A01783-2 is the only listed option. Its WB image provides an example in three human cell lines; consider the reported 37 kDa band and expected 29 kDa size when interpreting results.
Source: BosterBio MSX2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P35548.
- Human Protein Atlas. MSX2 tissue expression.
- PMC9953509 — target-verified WB comparison
- PMC4835074 — target-verified WB comparison
- PMC2949651 — target-verified WB comparison