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MTMR4 / Phosphatidylinositol-3,5-bisphosphate 3-phosphatase MTMR4 · Western blot design guide
Design a Western Blot for MTMR4
Source-linked MTMR4 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MTMR4 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
MTMR4 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 | ~133.4 kDa | |
| Gel | 8–10% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MTMR4 Western Blot Protocol Options
The A09354 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A09354) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A09354) |
| Primary antibody | A09354 · 1:1000 (catalog A09354) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A09354) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A09354) |
Section 2
What Is the Expected MTMR4 Western Blot Band Size?
Full-length MTMR4 is predicted at 133.4 kDa; phosphorylation and oligomerization could affect migration, but no empirical band or feature-driven shift is established.
| Band near 133.4 kDa | consistent with predicted full-length MTMR4; confirm identity with controls |
| Higher band | could reflect retained MTMR4 oligomers; identity needs confirmation |
| Faint or absent band in a soluble fraction | MTMR4 associates with endosome membranes |
| Close doublet near 133.4 kDa | could reflect different phosphorylation states; visible separation is unproven |
💡Expected MTMR4 appearanceUniProt predicts full-length MTMR4 at 133.4 kDa; no empirical band size is supplied, so confirm any candidate band with ordinary identity controls.
| Predicted full-length mass | sets a 133.4 kDa reference for band assessment |
| Phosphoserine at Ser8 | may alter apparent migration, but no shift is established |
| Phosphoserine at Ser610 | may alter apparent migration, but no shift is established |
| Phosphoserine at Ser629 | may alter apparent migration, but no shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | endosome-associated MTMR4 may be poorly recovered | check membrane-containing fractions and verify extraction |
| Band higher than expected | retained MTMR4 oligomers are possible | compare denaturing conditions and confirm band identity |
| Band lower than expected | the band may be a fragment or an unrelated protein; no cleavage feature is listed | check sample integrity and confirm identity with an independent antibody |
| Multiple bands | phosphorylation states are possible, but distinct migration is unproven | compare phosphatase-treated samples and confirm band identity |
| Weak or no signal | membrane-associated MTMR4 may be underrepresented in the tested fraction | check fraction recovery and include a positive control |
Experiment evidence
Western blot images showing MTMR4 band
A09354 · MTMR4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MTMR4 Western blot
🧪For positive controls for MTMR4 in Western blot, you can use no HPA-supported tissue or cell sample from the supplied evidence.
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 data, so suitable positive and negative samples cannot be identified from this evidence.
HPA tissue expression evidence for MTMR4
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 MTMR4 Western Blot Tips
Deeper troubleshooting and optimisation questions for MTMR4, answered from its protein features.
Where should the MTMR4 band appear relative to its predicted mass?
Band shift · The supplied predicted mass is 133.4 kDa. No empirical band position is provided, so use 133.4 kDa as a reference rather than an expected exact migration position.
Do listed MTMR4 isoforms suggest multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Could phosphorylation explain a shifted MTMR4 band?
PTM · UniProt lists phosphoserine at positions 8, 610, and 629. These features establish modification sites but do not show that phosphorylation produces a visible shift or explain any observed mass difference. The positions use UniProt numbering, which may differ from antibody or paper numbering.
For a phosphorylation-focused experiment, distinguish total MTMR4 from phosphorylation at UniProt positions 8, 610, or 629 when choosing what to measure. The supplied record lists these sites but gives no condition that induces their phosphorylation.
For a phosphorylation-focused experiment, distinguish total MTMR4 from phosphorylation at UniProt positions 8, 610, or 629 when choosing what to measure. The supplied record lists these sites but gives no condition that induces their phosphorylation.
Does this guide establish induction of MTMR4?
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.
What transfer method to use for MTMR4 Western blot?
Transfer · MTMR4 has a predicted mass of 133.4 kDa. Choose transfer conditions that recover proteins around that size, and check transfer efficiency before interpreting a weak or absent band. The supplied features do not specify a transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A09354 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 can sample fraction affect MTMR4 quantitation?
Quantitation · MTMR4 is listed as a peripheral protein at early and recycling endosome membranes. Keep fractionation and sample preparation consistent across samples, and compare signal within the same type of fraction when quantifying MTMR4.
How should unexpected MTMR4 bands be interpreted?
Interpretation · The record lists a 133.4-kDa protein, three phosphoserines, one isoform, and MTMR4 oligomers with MTMR3. None of these features alone identifies an unexpected band. Check band specificity and sample preparation before assigning it to an isoform, phosphorylation, or an oligomer.
Boster reagents
MTMR4 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-Myotubularin-related protein 4 MTMR4 Antibody
Cat # A09354
A09354 is an anti-MTMR4 antibody listed for human, mouse, and rat reactivity. Its Western blot image uses extracts from various cell lines at 1:1000 dilution with 25 µg per lane; the caption does not identify the cell lines.
Which to pick: A09354 is the only listed option and has a Western blot image. Check its reported species reactivity against your sample, and use the pictured 1:1000 dilution and 25 µg per lane as starting conditions.
Source: BosterBio MTMR4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NYA4.
- Human Protein Atlas. MTMR4 tissue expression.
- PMC2657291 — target-verified WB comparison