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MTSS1 / Protein MTSS 1 · Western blot design guide
Design a Western Blot for MTSS1
Source-linked MTSS1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MTSS1 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
MTSS1 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 | ~82.3 kDa | |
| Observed band | ~115 kDa | |
| Gel | 5–20% (catalog A04325-1) | |
| Positive control | Adrenal gland (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 | 4 isoform(s) |
Section 1
Source-Linked MTSS1 Western Blot Protocol Options
The A04325-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human placenta, rat spleen, rat thymus, mouse thymus (catalog A04325-1) |
| Gel % | 5–20% (catalog A04325-1) |
| Load | 30 ug; reducing conditions (catalog A04325-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04325-1) |
| Membrane | nitrocellulose membrane (catalog A04325-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04325-1) |
| Primary antibody | A04325-1 · 0.5 μg/mL (catalog A04325-1) |
| Primary incubation | overnight at 4°C (catalog A04325-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04325-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04325-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04325-1) |
| Detection | ECL (catalog A04325-1) |
Section 2
What Is the Expected MTSS1 Western Blot Band Size?
MTSS1 is predicted at 82.3 kDa but observed near 115 kDa; the cause of this difference is not established.
| Band near 115 kDa | Empirically observed MTSS1-reactive band; confirm identity with controls |
| Band near 82.3 kDa | Near the predicted sequence mass, but not the supplied empirical band |
| Several bands at different positions | Could reflect MTSS1 isoforms 1 through 4; their migration is unverified |
| Weak band in a soluble fraction | MTSS1 may remain with the cytoskeleton |
💡Expected MTSS1 appearanceMTSS1 has a predicted mass of 82.3 kDa and an empirical band near 115 kDa; the reason for the difference is unestablished, so confirm band identity with appropriate controls.
| Predicted MTSS1 mass | 82.3 kDa is the calculated sequence mass, not its demonstrated migration |
| Isoform 1 | May migrate differently from other named isoforms; its individual mass is unavailable |
| Isoform 2 | May migrate differently from other named isoforms; its individual mass is unavailable |
| Isoform 3 | May migrate differently from other named isoforms; its individual mass is unavailable |
| Isoform 4 | May migrate differently from other named isoforms; its individual mass is unavailable |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeleton-associated MTSS1 may be poorly extracted | Check extraction of the cytoskeletal fraction and include a positive control |
| Band higher than expected | The observed 115 kDa band exceeds the 82.3 kDa prediction for an unestablished reason | Compare with a positive control and confirm specificity by MTSS1 depletion |
| Band lower than expected | A different splice isoform may migrate at another position | Check isoform expression and confirm the band with MTSS1 depletion |
| Broad smear instead of sharp band | The cause is unestablished; MTSS1 has annotated phosphorylation sites | Compare treated and untreated samples and verify MTSS1 specificity |
| Multiple bands | MTSS1 has four named splice isoforms, though distinct bands are unverified | Compare isoform expression and test band specificity by MTSS1 depletion |
Experiment evidence
Western blot images showing MTSS1 band
A04325-1 · MTSS1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MTSS1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MTSS1 in Western blot, you can use adrenal gland lysate, which HPA rates High.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Caudate is an HPA Not-detected tissue, but confirm its lack of signal in your lysate.
HPA tissue expression evidence for MTSS1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Skin | fibroblasts | High | Protein (IHC) | HPA → |
| Small intestine | glandular cells | 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 → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced MTSS1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MTSS1, answered from its protein features.
How should MTSS1 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 could isoform 2 affect the band pattern?
Isoforms · Isoform 2 lacks residues 1–200 and 345–426 of the canonical sequence, so it may appear at a different position from full-length MTSS1. Check whether the antibody recognizes a region retained in isoform 2 before interpreting a missing or additional band.
Isoform 3 replaces canonical residues 346–409 with a different sequence. Isoform 4 changes the canonical residue at position 153 from K to KVDTL. An antibody targeting either affected region may recognize these isoforms differently; band position alone cannot identify them.
Match the epitope to the isoforms you intend to detect. Isoform 2 lacks canonical residues 1–200 and 345–426; isoform 3 replaces residues 346–409; isoform 4 alters position 153. An epitope within those regions may change which isoforms the antibody detects.
Isoform 3 replaces canonical residues 346–409 with a different sequence. Isoform 4 changes the canonical residue at position 153 from K to KVDTL. An antibody targeting either affected region may recognize these isoforms differently; band position alone cannot identify them.
Match the epitope to the isoforms you intend to detect. Isoform 2 lacks canonical residues 1–200 and 345–426; isoform 3 replaces residues 346–409; isoform 4 alters position 153. An epitope within those regions may change which isoforms the antibody detects.
Which MTSS1 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at 258, 425 and 603, and phosphoserine at 261, 262, 271, 322, 644 and 647. These are canonical UniProt coordinates; paper or antibody numbering may differ. The annotations establish modification sites, not that phosphorylation produces a resolvable band shift.
Does this guide establish induction of MTSS1?
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 MTSS1?
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 A04325-1 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 MTSS1 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 MTSS1 appear near 115 kDa instead of 82.3 kDa?
Interpretation · The supplied observed band is approximately 115 kDa, while the predicted mass is 82.3 kDa. MTSS1 has four isoforms and nine annotated phosphorylation sites, but these features alone do not explain the difference or establish a visible shift. Confirm the band’s identity before assigning a cause.
Quantify the same identified band across samples. MTSS1 has four annotated isoforms and nine phosphorylation sites, so pooling distinct bands could mix different species. Record which band was measured, especially when comparing signals near the reported 115 kDa position.
Alternative splicing provides one possibility: isoform 2 has two deleted regions, isoform 3 has a replaced region, and isoform 4 has a short sequence change. The listed phosphorylation sites are another feature to consider, but neither feature proves the identity of an unexpected band. Check isoform coverage and antibody epitope before assigning it.
Quantify the same identified band across samples. MTSS1 has four annotated isoforms and nine phosphorylation sites, so pooling distinct bands could mix different species. Record which band was measured, especially when comparing signals near the reported 115 kDa position.
Alternative splicing provides one possibility: isoform 2 has two deleted regions, isoform 3 has a replaced region, and isoform 4 has a short sequence change. The listed phosphorylation sites are another feature to consider, but neither feature proves the identity of an unexpected band. Check isoform coverage and antibody epitope before assigning it.
Boster reagents
MTSS1 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-MIM/MTSS1 Antibody Picoband®
Cat # A04325-1
The catalog reports one anti-MTSS1 antibody, A04325-1, with reported human, mouse, and rat reactivity. Its WB caption reports a band near 115 kDa, versus an expected 82 kDa. No publication evidence is supplied.
Which to pick: A04325-1 is the only listed option and has a WB image using human Jurkat cells and placenta, rat spleen and thymus, and mouse thymus. Consider the reported band-size discrepancy when interpreting results.
Source: BosterBio MTSS1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43312.
- Human Protein Atlas. MTSS1 tissue expression.
- PMC7952249 — target-verified WB comparison
- PMC9738320 — target-verified WB comparison
- PMC7477433 — target-verified WB comparison