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- Table of Contents
MNX1 / Motor neuron and pancreas homeobox protein 1 · Western blot design guide
Design a Western Blot for MNX1
Source-linked MNX1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MNX1 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
MNX1 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~40.6 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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked MNX1 Western Blot Protocol Options
The M04520 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Molt-4 cell lysate (catalog M04520) |
| 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 | M04520; 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 MNX1 Western Blot Band Size?
MNX1 is predicted at 40.6 kDa; isoforms and phosphorylation could affect migration, but no empirical band size or visible effect is established.
| Band near 40.6 kDa | consistent with the predicted MNX1 mass; confirm its identity with controls |
| Bands at different positions | could reflect isoforms 1 and 2, but their migration is not established |
| Band with altered mobility | could reflect phosphorylation at Ser77 or Ser79; a visible shift is not established |
| Weak band in a nuclear-poor lysate | consistent with MNX1 localization in the nucleus |
💡Expected MNX1 appearanceMNX1 has a predicted mass of 40.6 kDa, but no empirical band size is supplied; confirm a candidate band with appropriate identity controls because isoform and phosphorylation effects on migration are unestablished.
| Predicted molecular mass | places the reference size at 40.6 kDa |
| Splice isoforms 1 and 2 | may differ in size, but their relative sizes and migration are not supplied |
| N-acetylmethionine at residue 1 | adds a small modification without an established visible shift |
| Phosphoserine at residue 77 | may alter mobility; no shift is established |
| Phosphoserine at residue 79 | may alter mobility; no shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear MNX1 may be poorly recovered | check a nuclear fraction and verify extraction with a nuclear marker |
| Band higher than expected | an isoform or phosphorylation may affect migration, but neither effect is established | compare with a validated MNX1 control and use knockdown to test identity |
| Band lower than expected | an alternative isoform or degradation is possible | compare fresh lysate with a validated MNX1 control and use knockdown to test identity |
| Multiple bands | isoforms 1 and 2 or differing phosphorylation states are possible | use MNX1 knockdown to identify specific bands |
| Weak or no signal | nuclear MNX1 may be underrepresented in the sample | enrich the nuclear fraction and check the loading and nuclear markers |
Experiment evidence
Western blot images showing MNX1 band
M04520 · MNX1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MNX1 Western blot
🧪For positive controls for MNX1 in Western blot, you can use an HPA-IHC candidate positive sample once one is identified.
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 tissue or cell data here, so suitable positive and negative controls require independent validation.
HPA tissue expression evidence for MNX1
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 MNX1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MNX1, answered from its protein features.
How should MNX1 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.
How might MNX1 isoforms affect the band pattern?
Isoforms · UniProt lists two isoforms. Relative to the 401-residue canonical isoform 1, isoform 2 lacks residues 1–212 and has a replacement sequence at 213–231. It may produce a distinct band, but these features do not establish its apparent position on a blot.
No. Isoform 2 lacks canonical residues 1–212, which contain all three listed sites: acetylated position 1 and phosphorylated positions 77 and 79. An antibody directed at either listed phosphosite would therefore target isoform 1 rather than isoform 2, subject to antibody validation.
Check the epitope against both sequences. Canonical residues 1–212 are missing from isoform 2, and residues 213–231 are replaced. An epitope in those regions may distinguish the isoforms; an epitope in the shared sequence after residue 231 could detect both if the antibody recognizes each.
No. Isoform 2 lacks canonical residues 1–212, which contain all three listed sites: acetylated position 1 and phosphorylated positions 77 and 79. An antibody directed at either listed phosphosite would therefore target isoform 1 rather than isoform 2, subject to antibody validation.
Check the epitope against both sequences. Canonical residues 1–212 are missing from isoform 2, and residues 213–231 are replaced. An epitope in those regions may distinguish the isoforms; an epitope in the shared sequence after residue 231 could detect both if the antibody recognizes each.
Which MNX1 modifications matter when interpreting bands?
PTM · The listed modifications are N-acetylmethionine at position 1 and phosphoserines at positions 77 and 79, using canonical UniProt numbering. Their presence alone does not establish a visible band shift. Check numbering conventions before comparing antibody or paper site labels.
Does this guide establish induction of MNX1?
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 MNX1?
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 M04520 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 MNX1 bands be quantified across samples?
Quantitation · Decide whether the measurement is for one isoform or total MNX1, then use an antibody whose epitope fits that choice. MNX1 is listed as nuclear, so compare equivalent nuclear preparations and normalize consistently. Quantify distinct candidate isoform bands separately unless their identities have been verified.
Should MNX1 run at its predicted 40.6 kDa?
Interpretation · 40.6 kDa is the predicted mass for canonical MNX1. No observed band position is supplied, so a difference between predicted and apparent mass cannot be established or explained from these features alone.
Compare them with the two listed isoforms and the antibody epitope. Isoform 2 has a large N-terminal deletion and a replacement at canonical residues 213–231. The listed phosphorylation and acetylation sites do not, by themselves, explain an extra band or an apparent mass difference; band identity needs independent validation.
Compare them with the two listed isoforms and the antibody epitope. Isoform 2 has a large N-terminal deletion and a replacement at canonical residues 213–231. The listed phosphorylation and acetylation sites do not, by themselves, explain an extra band or an apparent mass difference; band identity needs independent validation.
Boster reagents
MNX1 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-HB9/HLXB9 Monoclonal Antibody
Cat # M04520
The catalog reports one anti-MNX1 antibody, M04520 (Anti-HB9/HLXB9), with reported Human and Mouse reactivity. Its Western blot image shows HB9/HLXB9 expression in Molt-4 cell lysate; the supplied evidence does not show a Mouse WB sample.
Which to pick: M04520 is the only listed option and has a WB image from Molt-4 cell lysate. For Mouse samples, check the reported reactivity against your application needs, since no Mouse WB example is supplied.
Source: BosterBio MNX1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P50219.
- Human Protein Atlas. MNX1 tissue expression.
- PMC6102707 — target-verified WB comparison
- PMC7581777 — target-verified WB comparison
- PMC12732208 — target-verified WB comparison