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MTSS1 / Protein MTSS 1 · IHC design guide
Design Immunohistochemistry for MTSS1
Plan chromogenic MTSS1 IHC on paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A04325-1). Use kidney tubules and duodenal glandular cells as high-staining reference tissues, and assess the cytoplasmic pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MTSS1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic staining, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04325-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Caudate+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression varies by tissue and cell type (HPA tissue IHC) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended MTSS1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A04325-1) with four published MTSS1 paraffin-section protocols (PMC3492170; PMC4453956; PMC6006423; PMC3131255).
| Sample | Paraffin-embedded human endometrial cancer tissue; fixative not specified (datasheet A04325-1) |
| Fixation | Image fixative and duration unreported (datasheet A04325-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04325-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04325-1) |
| Primary antibody | Rabbit anti-MTSS1, 2-5 μg/ml (datasheet A04325-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04325-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A04325-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MTSS1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression of variable levels in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet A04325-1). Published alternatives include pH 9 retrieval and pH 6.0 citrate (PMC4453956; PMC3131255).
Section 2
What Is the Expected MTSS1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic MTSS1 staining, with intensity varying by cell type (HPA: cytoplasmic expression at variable levels in most tissues). Glandular cells in duodenum and small intestine, kidney tubule cells, and testis spermatogonia are strong reference populations (HPA: High). MTSS1 is associated with the cytoplasm and cytoskeleton and has no transmembrane segment (UniProt O43312). HPA rates its tissue IHC profile Approved, with medium consistency against RNA data and external verification pending (HPA: reliability description).
| Clear cytoplasmic chromogen in duodenal glandular cells or kidney tubule cells. | This matches the expected compartment and cell populations (HPA: cytoplasmic profile; High in duodenal glandular cells and kidney tubule cells). Score the named cells separately from neighboring tissue; a positive control establishes that the staining run worked, while the antibody still needs specificity controls (general IHC practice). |
| Predominantly nuclear, luminal, or broadly extracellular staining. | Treat a pattern dominated by these compartments as suspect: they are outside the reported cytoplasmic tissue profile (HPA: tissue IHC) and cytoplasm/cytoskeleton annotation (UniProt O43312). Membrane accent alone is less decisive because plasma membrane and cytosol were observed by ICC-IF (HPA: subcellular); that IF observation does not establish a required membrane pattern in paraffin IHC. |
| Strong chromogen in a population recorded as undetected, such as alveolar cells. | Investigate possible cross-reactivity or endogenous detection activity before calling MTSS1 positive (HPA: Not detected in lung alveolar cells; general IHC practice). The HPA designation applies to that cell population, not to every cell in the section; compare morphology and controls before assigning the source of signal. |
| Diffuse color across cells and blank spaces, with little separation between cell populations. | This is difficult to score as MTSS1 because the reference is a cellular, variable cytoplasmic pattern (HPA: tissue IHC). Background can arise from nonspecific binding or detection chemistry (general IHC practice). Review a no-primary control and whether the chromogen obscures the counterstain before interpreting weak cells. |
| No discernible staining in duodenal glandular cells or kidney tubule cells. | Do not interpret a fully negative run until a known-positive population and run controls have been checked (HPA: High in these cells; general IHC practice). Review antibody and detection setup, then assess retrieval using the validated IHC-P procedure; the supplied sources report no MTSS1-specific fixation or retrieval sensitivity. |
💡Expected MTSS1 appearanceCall a section positive when identifiable glandular cells, kidney tubule cells, or spermatogonia show cell-associated, mainly cytoplasmic chromogen, potentially High in the listed reference populations (HPA: tissue IHC); dominant nuclear or extracellular color, or uniform color across named positive and undetected populations, warrants investigation rather than an MTSS1 call (HPA: cytoplasmic profile; general IHC practice).
| Cell and tissue context | Intensity varies across tissues and cell populations (HPA: variable cytoplasmic expression; low tissue RNA specificity). HPA records High staining in adrenal and intestinal glandular cells, kidney tubule cells, testis spermatogonia, and tonsil non-germinal center cells; it records Not detected in cardiomyocytes and smooth muscle cells (HPA: tissue IHC). Compare like cells, not whole-section averages. |
| Compartment and topology | A cytoplasmic/cytoskeletal result is consistent with the protein annotation (UniProt O43312: subcellular location). MTSS1 has no transmembrane segment or signal peptide (UniProt O43312: topology and processing). These annotations help assess localization; they do not predict how fixation or antigen retrieval will affect a particular antibody. |
| Antibody validation | HPA lists IHC Supported for HPA064003 and IHC Approved for HPA075540; it provides no IHC status for HPA019613 (HPA: antibody validation). The overall tissue profile is Approved but awaits external verification (HPA: reliability description). Keep the validation status tied to the antibody used; the record does not establish IHC Enhanced validation. |
| Isoforms and epitope scope | UniProt lists 4 MTSS1 isoforms and IMD and WH2 domains (UniProt O43312). The payload gives no antibody epitope, so it cannot establish which isoforms an IHC antibody detects or predict a compartment change from epitope position. Record the antibody identity when comparing results (general IHC practice). |
| IF/ICC interpretation — what should appear? | HPA reports approved plasma-membrane and cytosol localization in ICC-IF and lists HEL among imaged cell lines (HPA: subcellular). That supports a separate IF/ICC localization comparison, not an IHC-P protocol choice or a requirement for membrane staining in chromogenic paraffin sections. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-control glandular cells or kidney tubule cells remain blank. | A failed detection run or insufficient signal is possible (general IHC practice); these cells are reported High (HPA: tissue IHC). | Check the known-positive section, primary and detection reagents, and counterstain; verify the IHC-P antibody procedure and assess retrieval within that procedure (general IHC practice). The supplied sources do not identify an MTSS1-specific retrieval condition. |
| The section looks uniformly brown, including areas without identifiable target cells. | Nonspecific reagent binding or detection background may obscure a variable cellular pattern (general IHC practice; HPA: cytoplasmic profile). | Review a no-primary control, washing, blocking, and chromogen development (general IHC practice). Score MTSS1 only where cell borders and cytoplasm can be distinguished; do not convert diffuse background into a tissue-wide positive call. |
| Unexpected color appears in alveolar cells or cardiomyocytes. | Those populations are listed Not detected (HPA: lung alveolar cells; heart cardiomyocytes); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm cell identity and inspect a no-primary control; for peroxidase detection, assess endogenous peroxidase blocking (general IHC practice). If signal persists only with primary antibody, investigate specificity before reporting it as MTSS1. |
| Most color appears nuclear or in luminal material. | That distribution conflicts with the reported cytoplasmic tissue profile (HPA: tissue IHC) and cytoplasm/cytoskeleton annotation (UniProt O43312). | Check morphology, controls, and whether a cytoplasmic component is present (general IHC practice). Do not score nuclear or luminal color alone as the expected MTSS1 result; a narrow membrane accent can be assessed separately (HPA: ICC-IF membrane and cytosol). |
| One tissue stains weakly while another gives a clear positive result. | A true difference between cell populations is plausible (HPA: variable cytoplasmic levels; Low in thyroid glandular cells; High in duodenal glandular cells). | Compare the same named cell type and a known-positive section, then score intensity by population (general IHC practice). Avoid treating every weak area as a failed run when the positive control and cellular pattern are intact. |
| An IF/ICC image shows membrane signal that seems absent from the IHC slide. | The modalities have different reported observations: HPA describes plasma membrane and cytosol in ICC-IF and variable cytoplasmic staining in tissue IHC (HPA: subcellular and tissue IHC). | Interpret each assay against its own reported pattern; use the separate IF/ICC guide for that application. A membrane signal in IF does not, by itself, make predominantly cytoplasmic chromogenic IHC incorrect (HPA: subcellular and tissue IHC). |
Experiment evidence
IHC & IF images showing MTSS1 staining
A04325-1 · IHCIHC analysis of MIM/MTSS1 using anti-MIM/MTSS1 antibody (A04325-1). MIM/MTSS1 was detected in a paraffin-embedded section of human endometri
A04325-1 · IHCIHC analysis of MIM/MTSS1 using anti-MIM/MTSS1 antibody (A04325-1). MIM/MTSS1 was detected in a paraffin-embedded section of human gastric c
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000170873-MTSS1/tissue/cerebral+cortex#imid_20074389
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000170873-MTSS1/tissue/colon#imid_20074307
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000170873-MTSS1/tissue/liver#imid_20074308
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000170873-MTSS1/tissue/kidney#imid_20074317
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA019613; green = MTSS1, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA019613; green = MTSS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MTSS1 IHC & IF
🧪Run kidney first: tubular cells should stain strongly (HPA: High in kidney cells in tubules). Use heart muscle as a negative tissue, where cardiomyocytes are not detected (HPA: Not detected in cardiomyocytes); on the kidney slide, cells without specific staining should show counterstain without DAB signal (standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MTSS1 in HEL, with annotated localisation: Plasma membrane (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and rabbit isotype controls, matching the primary antibody’s class, clonality, and concentration where known (caption: rabbit primary antibody; standard IHC practice). Use validated MTSS1-knockout material as a biological negative if available, and block endogenous peroxidase before HRP–DAB detection; if using a biotin-based system in kidney, assess endogenous biotin (caption: HRP–DAB detection; standard IHC practice).
⚠️Feasibility: The selected A04325-1 paraffin-section caption reports heat retrieval in EDTA at pH 8.0 and a primary concentration of 2 μg/ml, but its fixative is unreported (selected-SKU tissue-IHC caption). A target-specific fixation window or fixation effect is unreported, and the supplied evidence does not establish whether frozen sections or IF are easier (supplied target/application evidence). For IF/ICC, what localisation should be checked? Plasma membrane and cytosol staining is the reported pattern in HEL images; assess kidney tubular autofluorescence with a no-primary control if using that tissue for IF (HPA subcellular; standard IF practice).
HPA tissue IHC evidence for MTSS1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended 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 → |
Section 3
Advanced MTSS1 IHC Tips
Troubleshoot MTSS1 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
What retrieval conditions should I try when MTSS1 staining is weak?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04325-1). The illustrated section used this retrieval before 2 μg/ml primary antibody overnight at 4°C, so keep those conditions together for the first comparison (datasheet A04325-1). If staining remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration, detection and development constant (standard IHC practice). Check a section containing kidney tubules or duodenal glandular cells, where high staining is reported, alongside the study specimen (HPA tissue IHC). Excessive retrieval can damage morphology and complicate compartment scoring, so judge signal and tissue preservation together (standard IHC practice).
How should I troubleshoot inconsistent MTSS1 staining after fixation?
The illustrated MTSS1 section is described as paraffin embedded, but its fixative is unreported; target-specific fixation sensitivity therefore remains unknown (datasheet A04325-1). Record fixative, fixation duration and processing history for each block before comparing staining across specimens (standard IHC practice). Process matched sections together, then compare morphology and MTSS1 signal using the same EDTA pH 8.0 retrieval and detection settings (datasheet A04325-1; standard IHC practice). Include an internal or companion positive control containing a reported high-staining cell population, such as kidney tubules (HPA tissue IHC). If signal varies with processing, report that association without assigning a specific MTSS1 fixation mechanism that has not been tested (standard IHC practice).
Where should MTSS1 signal appear in chromogenic tissue sections?
Evaluate MTSS1 primarily in the cytoplasmic compartment: tissue staining is reported as variably cytoplasmic, and the protein is associated with the cytoskeleton (HPA tissue IHC; UniProt O43312). Plasma membrane and cytosol localisation in cell imaging can help interpret peripheral staining, but that observation does not establish a tissue-specific staining pattern (HPA subcellular). MTSS1 has no transmembrane segment, so a sharp membrane outline alone warrants review of morphology and background controls (UniProt O43312 topology; standard IHC practice). Score the expected cell population separately from neighbouring cells because staining levels vary among tissues (HPA tissue IHC). Confirm any unexpected nuclear-only or extracellular deposit pattern with an independent control before calling it MTSS1 (standard IHC practice).
Could isoforms or epitope accessibility explain discordant MTSS1 staining?
MTSS1 has 4 annotated isoforms and an actin-related architecture that includes IMD residues 1–250 and WH2 residues 727–744 (UniProt O43312). The supplied tissue caption does not identify the antibody epitope, so it cannot establish which isoforms the catalog antibody detects (datasheet A04325-1). Ask for the immunogen or epitope map before attributing a negative section to isoform loss (standard IHC practice). Several phosphorylated residues are annotated, including positions 258 and 261, but their effect on this antibody's tissue staining is unestablished (UniProt O43312; datasheet A04325-1). Compare adjacent sections under identical retrieval and detection conditions before proposing epitope accessibility as the explanation (standard IHC practice).
How can I check an MTSS1 IHC finding with multiplex immunofluorescence?
For a separate IF experiment, pair MTSS1 with a validated marker identifying the cell population being scored in the chromogenic section (standard IF practice). Kidney tubule cells and duodenal glandular cells are reported high-staining reference populations, while cell imaging places MTSS1 at the plasma membrane and in cytosol (HPA tissue IHC; HPA subcellular). Choose spectrally separated fluorophores and measure unstained tissue autofluorescence before assigning faint MTSS1 signal, particularly in the brighter channel (standard IF practice). Because MTSS1 has no transmembrane segment, use permeabilisation when testing an intracellular epitope, adjusting it to preserve cell boundaries and the marker signal (UniProt O43312 topology; standard IF practice). The paraffin-section caption does not establish IF fixation or antibody performance (datasheet A04325-1).
What controls help distinguish MTSS1 staining from chromogenic background?
Run a no-primary control through the same secondary, peroxidase and DAB steps to assess detection background (standard IHC practice). The illustrated section used 10% goat serum blocking, a peroxidase-conjugated secondary and DAB development; treat these as documented conditions for SKU A04325-1, not proof of specificity (datasheet A04325-1). Include an appropriate peroxidase block and inspect unstained or control sections for endogenous enzyme signal (standard IHC practice). High staining reported in adrenal glandular cells can serve as a positive reference, while alveolar cells were reported as undetected in the surveyed lung tissue (HPA tissue IHC). Reassess diffuse edge staining or pigment before increasing primary concentration (standard IHC practice).
How should I quantify MTSS1 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since MTSS1 tissue staining is variably cytoplasmic and can differ by tissue (HPA tissue IHC). For a comparable cohort, record percentage of positive target cells and intensity categories, then calculate an H-score using the same thresholds for every section (standard IHC practice). If counting discrete positive cells, report density per mm² of viable target tissue and state the area-selection rule (standard IHC practice). Normalize each specimen to its eligible target-cell count or viable target-tissue area, while documenting batch and exposure or DAB development settings (standard IHC practice). Keep stromal, glandular and other compartments separate rather than letting their proportions drive the specimen score (standard IHC practice).
When is apparent MTSS1 positivity more likely to be artefact?
A credible result places signal in the scored cells with mainly cytoplasmic distribution, consistent with tissue observations and MTSS1 cytoskeletal association (HPA tissue IHC; UniProt O43312). Review nuclear-only staining, extracellular precipitate and isolated sharp membrane outlines against morphology and a no-primary control before assigning MTSS1 positivity (HPA subcellular; UniProt O43312 topology; standard IHC practice). Exclude section edges, folds and necrotic regions from scoring when they show disproportionate DAB deposition (standard IHC practice). Check endogenous peroxidase background with the relevant detection control (standard IHC practice). HPA rates its tissue IHC profile Approved but reports medium agreement with RNA expression and pending external verification, so unexpected patterns need independent corroboration (HPA tissue IHC).
Boster reagents
Best MTSS1 / Protein MTSS 1 IHC Antibodies
A04325-1 has chromogenic IHC images from paraffin-embedded human endometrial cancer, gastric carcinoma, lymphoma, and placenta sections (catalog image captions); IF/ICC data are unreported (catalog applications and images).
Real IHC data
Anti-MIM/MTSS1 Antibody ®
Cat # A04325-1
A04325-1 is the only SKU shown; its IHC images use paraffin-embedded human endometrial cancer, gastric carcinoma, lymphoma, and placenta sections (catalog image captions). Its listed reactivity is human, mouse, and rat, while its IHC dilution is specified for human samples (catalog: reactivity and dilution).
Which to pick: Choose A04325-1 for human tissue IHC: its caption documents heat retrieval in EDTA at pH 8.0 and primary antibody at 2 μg/ml on a paraffin section; the fixative is unreported (A04325-1 image caption). No IF/ICC SKU is validated here (catalog: applications and IF images). A04325-1 lists mouse and rat reactivity, but the supplied IHC images show human tissue only; clone information is unreported (catalog: reactivity, image captions, clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43312 (MTSS1_HUMAN, Protein MTSS 1).
- Human Protein Atlas. MTSS1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MTSS1 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. MTSS1 antibody validation summary (3 antibodies).
- MicroRNA-182 downregulates metastasis suppressor 1 and contributes to metastasis of hepatocellular carcinoma. BMC cancer 2012 — PMC3492170.
- Downregulation of MTSS1 expression is an independent prognosticator in squamous cell carcinoma of the lung. British journal of cancer 2015 — PMC4453956.
- Expression and clinical significance of miR-23a and MTSS1 in diffuse large B-cell lymphoma. Oncology letters 2018 — PMC6006423.
- The impact of Metastasis Suppressor-1, MTSS1, on oesophageal squamous cell carcinoma and its clinical significance. Journal of translational medicine 2011 — PMC3131255.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MTSS1 (Protein MTSS 1). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04325-1). Validated IHC antibody: A04325-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human endometrial cancer tissue; fixative not specified (datasheet A04325-1); 2. Fixation: Image fixative and duration unreported (datasheet A04325-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A04325-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04325-1); 8. Primary antibody: Rabbit anti-MTSS1, 2-5 μg/ml (datasheet A04325-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04325-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 4 isoforms; epitope coverage is unknown (UniProt); Regulation: Expression varies by tissue and cell type (HPA tissue IHC). Evidence: UniProt O43312, HPA tissue IHC · Approved, HPA ICC-IF · plasma membrane and cytosol., Datasheet A04325-1, PMC3492170, PMC4453956. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MTSS1/immunohistochemistry-ihc-guide.