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- Table of Contents
MXI1 / Max-interacting protein 1 · Western blot design guide
Design a Western Blot for MXI1
Source-linked MXI1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MXI1 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
MXI1 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 | ~26.1 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked MXI1 Western Blot Protocol Options
The A08708 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of Mouse brain, (catalog A08708) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A08708) |
| Primary antibody | A08708 · 1:1000 (catalog A08708) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A08708) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A08708) |
Section 2
What Is the Expected MXI1 Western Blot Band Size?
MXI1 is predicted at 26.1 kDa; four isoforms could affect migration, but no distinct band pattern or empirical size is demonstrated.
| Single band near 26.1 kDa | Consistent with the predicted MXI1 mass; confirm identity with antibody controls |
| Several bands at different positions | Could reflect isoforms 1, 2, 3, and 4; their migration differences are unverified |
| Weak band in whole-cell lysate | Nuclear localization may dilute MXI1 signal in this sample |
| Little or no band in a cytosolic fraction | Consistent with MXI1 localization in the nucleus |
💡Expected MXI1 appearanceMXI1 has a predicted mass of 26.1 kDa, but no empirical band size is supplied; use antibody specificity controls to identify any band near that position or other bands.
| Predicted MXI1 mass | Places the reference band near 26.1 kDa |
| Isoform 1 | May migrate differently from other isoforms; its mass is unspecified |
| Isoform 2 | May migrate differently from other isoforms; its mass is unspecified |
| Isoform 3 | May migrate differently from other isoforms; its mass is unspecified |
| Isoform 4 | May migrate differently from other isoforms; its mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear MXI1 may be poorly represented in the sampled lysate | Check a nuclear extract and verify antibody performance with a positive control |
| Band higher than expected | An isoform or an unrelated protein may account for the band; its identity is unestablished | Compare with an MXI1 knockdown or knockout control |
| Band lower than expected | An isoform or an unrelated protein may account for the band; its identity is unestablished | Compare with an MXI1 knockdown or knockout control |
| Multiple bands | Four named isoforms are reported, but distinct migration is unverified | Use an MXI1 knockdown or knockout control to identify specific bands |
| Weak or no signal | Nuclear localization may limit MXI1 abundance in the tested fraction | Test a nuclear extract alongside a positive control |
Experiment evidence
Western blot images showing MXI1 band
A08708 · MXI1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MXI1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MXI1 in Western blot, you can use appendix tissue, where HPA reports high expression.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: MXI1 is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for MXI1
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | 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 → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced MXI1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MXI1, answered from its protein features.
How should MXI1 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 MXI1 isoforms produce different bands?
Isoforms · Yes. Four isoforms are listed, with differences in their N-terminal sequences and, for isoform 4, residues 68..77. These differences make distinct band positions possible, but the supplied features do not establish their apparent masses.
Both lack canonical residues 1..36. Isoform 4 also lacks residues 68..77. These are UniProt canonical-sequence coordinates; compare the antibody's target region with these changes when assessing a band.
In isoform 3, canonical residues 1..25 are replaced by a different, longer N-terminal sequence. An antibody targeting that region may recognize isoform 3 differently. The supplied features do not predict its apparent band position.
Both lack canonical residues 1..36. Isoform 4 also lacks residues 68..77. These are UniProt canonical-sequence coordinates; compare the antibody's target region with these changes when assessing a band.
In isoform 3, canonical residues 1..25 are replaced by a different, longer N-terminal sequence. An antibody targeting that region may recognize isoform 3 differently. The supplied features do not predict its apparent band position.
Does UniProt identify a modification that explains an MXI1 band shift?
PTM · No modified residues or glycosylation sites are listed in the supplied features. They therefore provide no specific modification or site to assign to a shifted band; they also do not establish the cause of any observed shift.
Does this guide establish induction of MXI1?
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 MXI1?
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 A08708 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 MXI1 when multiple bands appear?
Quantitation · Choose and report the band or bands being measured consistently. MXI1 has four annotated isoforms, so a single band should not automatically be treated as total MXI1. The supplied features do not identify which apparent band corresponds to each isoform.
What molecular weight should I expect for MXI1?
Interpretation · The supplied predicted mass is 26.1 kDa. No observed band position is available, so this value alone cannot establish where MXI1 will migrate on a Western blot.
Check whether its interpretation is consistent with the annotated isoform changes: loss of residues 1..36 in isoforms 2 and 4, loss of 68..77 in isoform 4, or replacement of 1..25 in isoform 3. Sequence differences alone cannot identify an unexpected band or prove a visible shift.
Check whether its interpretation is consistent with the annotated isoform changes: loss of residues 1..36 in isoforms 2 and 4, loss of 68..77 in isoform 4, or replacement of 1..25 in isoform 3. Sequence differences alone cannot identify an unexpected band or prove a visible shift.
Boster reagents
MXI1 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-Max-interacting protein 1 MXI1 Antibody
Cat # A08708
A08708 is an anti-MXI1 antibody listed as reactive with human, mouse, and rat. Its supplied Western blot image uses mouse brain extract at 1:1000 dilution; the provided evidence does not show Western blot results for human or rat samples.
Which to pick: Only A08708 is listed. It has a Western blot image from mouse brain extract (25 µg per lane), making it the documented choice for that sample. Human and rat reactivity is listed, but no Western blot images for those species are supplied.
Source: BosterBio MXI1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P50539.
- Human Protein Atlas. MXI1 tissue expression.
- PMC4350643 — target-verified WB comparison
- PMC2375210 — target-verified WB comparison
- PMC9061846 — target-verified WB comparison