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- Table of Contents
MTM1 / Myotubularin · Western blot design guide
Design a Western Blot for MTM1
Source-linked MTM1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MTM1 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
MTM1 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 | ~69.9 kDa | |
| Observed band | ~65 kDa | |
| Gel | 10% (catalog A00749-1) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked MTM1 Western Blot Protocol Options
The A00749-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human THP-1, human HepG2, rat heart, rat liver, mouse heart, mouse liver (catalog A00749-1) |
| Gel % | 10% (catalog A00749-1) |
| Load | 30 ug; reducing conditions (catalog A00749-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00749-1) |
| Membrane | nitrocellulose membrane (catalog A00749-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00749-1) |
| Primary antibody | A00749-1 · 0.5 μg/mL (catalog A00749-1) |
| Primary incubation | overnight at 4°C (catalog A00749-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00749-1) |
| Secondary incubation | 1.5 hour at RT (catalog A00749-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00749-1) |
| Detection | ECL (catalog A00749-1) |
Section 2
What Is the Expected MTM1 Western Blot Band Size?
MTM1 is predicted at 69.9 kDa and observed at ~65 kDa; the cause of this difference is not established.
| Band at ~65 kDa | Empirical MTM1 band in reducing lysates; its difference from the predicted mass is unexplained |
| Band near 69.9 kDa | Position consistent with the predicted MTM1 mass, requiring an identity control |
| Additional bands | Isoforms 1 and 2 are possible contributors, but distinct migration is unproven |
| Weak signal in a subcellular fraction | MTM1 may be distributed between cytoplasm and peripheral membrane |
💡Expected MTM1 appearanceMTM1 is predicted at 69.9 kDa, while antibody QC detects ~65 kDa in reducing lysates; the difference is unexplained, so confirm band identity with knockdown or an independent antibody.
| Predicted MTM1 mass | 69.9 kDa by sequence; the empirical band is ~65 kDa |
| Isoform 1 | individual mass and migration are unspecified |
| Isoform 2 | individual mass and migration are unspecified |
| Alternative splicing | may alter size, but no isoform-specific band position is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MTM1 recovery may depend on cytoplasmic and peripheral membrane extraction | compare whole-cell lysate with cytoplasmic and membrane fractions |
| Band higher than expected | identity or migration of a higher band is unestablished | check size markers and confirm with MTM1 knockdown or an independent antibody |
| Band lower than expected | the observed ~65 kDa band is below the 69.9 kDa prediction for an unknown reason | compare with the QC band and confirm with MTM1 knockdown |
| Multiple bands | isoforms 1 and 2 are possible, but separate band positions are unknown | test which bands respond to MTM1 knockdown |
| Weak or no signal | MTM1 may be poorly recovered from the sampled fraction | check extraction and compare cytoplasmic and membrane fractions |
Experiment evidence
Western blot images showing MTM1 band
A00749-1 · MTM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MTM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MTM1 in Western blot, you can use appendix tissue lysate, which HPA scores as high for MTM1.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: MTM1 is intracellular, and HPA identifies a not-detected tissue for a negative control.
HPA tissue expression evidence for MTM1
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 → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MTM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MTM1, answered from its protein features.
How should MTM1 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 MTM1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 78–114 relative to the canonical sequence, so it may migrate differently. Whether both isoforms are present or resolve as separate bands depends on the sample and assay.
Which MTM1 phosphorylation sites matter when interpreting bands?
PTM · The listed sites are phosphoserine 13, phosphoserine 18, phosphothreonine 495, and phosphoserine 588. These are UniProt canonical-sequence coordinates; antibody or paper numbering may differ. Their presence does not establish a detectable mobility shift.
Phosphorylation is one possibility because four modified residues are listed, but the features do not show that phosphorylation produces a second band. Compare samples with an appropriate phosphatase treatment if testing that explanation, and interpret the result alongside the two listed isoforms.
Phosphorylation is one possibility because four modified residues are listed, but the features do not show that phosphorylation produces a second band. Compare samples with an appropriate phosphatase treatment if testing that explanation, and interpret the result alongside the two listed isoforms.
Does this guide establish induction of MTM1?
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 MTM1?
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 A00749-1 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 MTM1 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.
Why might MTM1 appear near 65 kDa instead of 69.9 kDa?
Interpretation · The reported band is approximately 65 kDa, while the predicted mass is 69.9 kDa. These values alone do not establish the cause. Check whether the band is reproducible and whether its intensity tracks MTM1 abundance; the listed phosphorylation sites do not establish a visible shift.
Use the same sample fraction and loading basis across comparisons. MTM1 is listed in the cytoplasm and as a peripheral cell-membrane protein, with endosome and cell-projection annotations; changes in fraction recovery could change band intensity without establishing a change in total MTM1.
First compare them with the approximately 65 kDa reported band and consider isoform 2, which lacks canonical residues 78–114. The listed phosphorylation sites offer another hypothesis, but neither feature identifies an unexpected band on its own. MTM1 also forms a heterodimer with MTMR12; the interaction alone does not assign a band identity.
Use the same sample fraction and loading basis across comparisons. MTM1 is listed in the cytoplasm and as a peripheral cell-membrane protein, with endosome and cell-projection annotations; changes in fraction recovery could change band intensity without establishing a change in total MTM1.
First compare them with the approximately 65 kDa reported band and consider isoform 2, which lacks canonical residues 78–114. The listed phosphorylation sites offer another hypothesis, but neither feature identifies an unexpected band on its own. MTM1 also forms a heterodimer with MTMR12; the interaction alone does not assign a band identity.
Boster reagents
MTM1 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-MTM1 Antibody Picoband®
Cat # A00749-1
The catalog reports one anti-MTM1 antibody, A00749-1, with a Western blot image using human cell, rat tissue, and mouse tissue lysates. The reported band is approximately 65 kDa versus an expected 70 kDa. No independent validation is supplied.
Which to pick: A00749-1 is the only listed option. Its WB image includes human Jurkat, THP-1, and HepG2 cells and rat and mouse heart and liver tissue, using 30 µg per lane under reducing conditions. Match your sample and protocol to these reported contexts.
Source: BosterBio MTM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13496.
- Human Protein Atlas. MTM1 tissue expression.
- PMC4105197 — target-verified WB comparison
- PMC8851856 — target-verified WB comparison
- PMC9462472 — target-verified WB comparison