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- Table of Contents
Plan paraffin-section MTMR3 IHC around cytoplasmic staining, most abundant in seminiferous duct basal cells (HPA tissue IHC). Start with 2–5 μg/ml of catalog antibody A05937-1 (datasheet A05937-1), and compare cell-specific staining with a no-primary control (standard IHC practice).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in most tissues; strongest in seminiferous duct basal cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05937-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A05937-1); verify before use. | |
| Caveat | Tissue staining has uncertain concordance with RNA expression (HPA tissue IHC) | |
| Regulation | Bone marrow RNA is tissue enhanced (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; epitope coverage unknown; no transmembrane segment (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet A05937-1) is accompanied by published protocols for breast tumors (PMC6775797) and gastric cancer patient and mouse tumor samples (PMC7670601).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A05937-1) |
| Fixation | Image fixative and duration unreported (datasheet A05937-1); 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 A05937-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05937-1) |
| Primary antibody | Rabbit anti-MTMR3, 2-5 μg/ml (datasheet A05937-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05937-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05937-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MTMR3-positive staining in myoepithelial cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, most abundant in basal cells in seminiferous ducts. No signal in the no-primary control. |
MTMR3 is reported in the cytosol and at membranes, with no transmembrane segment (UniProt Q13615: subcellular location and topology). In tissue sections, expect mainly cytoplasmic staining; HPA reports high staining in cells of seminiferous ducts, bronchial respiratory epithelium and breast myoepithelium (HPA: tissue IHC). Treat this as a provisional pattern because HPA rates the tissue IHC profile uncertain (HPA: tissue IHC reliability).
| Cytoplasmic staining in basal cells of seminiferous ducts, respiratory epithelial cells or breast myoepithelial cells (HPA: tissue IHC). | This matches the reported tissue pattern; compare the named cell population with nearby cells, rather than scoring the entire section as uniformly positive (HPA: tissue IHC; general IHC practice). |
| A sharp membrane-only rim or exclusively nuclear chromogenic signal, with no discernible cytoplasmic staining (HPA: tissue IHC; UniProt Q13615: topology). | This is discordant with the reported IHC pattern and warrants an artefact check (HPA: tissue IHC; general IHC practice). Membrane association is possible, but UniProt reports no transmembrane segment (UniProt Q13615: subcellular location and topology). |
| Strong staining in adipocytes or liver cholangiocytes (HPA: not detected in those cell types). | This conflicts with the listed HPA observations for those specific cells. Check antibody specificity or endogenous detection activity before calling it MTMR3; neither observation makes its whole tissue a negative control (HPA: tissue IHC; general IHC practice). |
| Diffuse colour across cells and surrounding tissue, without recognizable cell boundaries (general IHC practice). | A diffuse deposit is difficult to score as a cellular pattern. Review blocking, washing, antibody concentration and chromogen development alongside a reagent control (general IHC practice). |
| No staining in cells of seminiferous ducts, bronchial respiratory epithelium or breast myoepithelium (HPA: high in these cell populations). | Check whether the expected cells are present, then assess section quality, retrieval and detection with appropriate controls (general IHC practice). HPA's uncertain IHC reliability limits what one negative section proves (HPA: tissue IHC reliability). |
| How firm is the reference pattern? (HPA: tissue IHC reliability) | HPA rates tissue IHC reliability uncertain, describing medium consistency between antibody staining and RNA expression; its two listed antibodies, HPA034515 and HPA034516, each have uncertain IHC validation (HPA: tissue IHC and antibody validation). |
| What compartment should guide scoring? (UniProt Q13615: subcellular location and topology) | UniProt lists cytosol and membrane and no transmembrane segment; HPA describes predominantly cytoplasmic tissue staining. Score the observed cellular distribution rather than requiring a continuous membrane outline (UniProt Q13615: subcellular location and topology; HPA: tissue IHC). |
| Can RNA abundance choose the strongest protein control? (HPA: tissue IHC) | HPA calls RNA expression tissue enhanced in bone marrow, yet lists bone-marrow hematopoietic cells at medium protein staining. Choose a positive cell population from the IHC observations, and keep RNA and protein readouts distinct (HPA: tissue IHC). |
| Do isoforms change the interpretation? (UniProt Q13615: isoforms) | UniProt lists 3 isoforms, B, A and C. The supplied sources give no antibody epitope mapping or isoform-specific tissue pattern, so these records cannot assign a staining difference to one isoform (UniProt Q13615: isoforms; HPA: antibody validation). |
| Q: Should IF/ICC show the same compartment as tissue IHC? (HPA: tissue IHC and subcellular ICC-IF) | A: HPA lists nucleoplasm as the main ICC-IF location with uncertain support, and cytosol as an additional supported location. Interpret that assay-specific observation separately from HPA's cytoplasmic tissue IHC pattern (HPA: subcellular ICC-IF and tissue IHC). |
| Could the detection system create colour? (general IHC practice) | Endogenous enzyme activity or nonspecific reagent binding can mimic chromogenic staining. Appropriate reagent controls help distinguish detection artefact from a cell-specific antibody signal (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are absent from the section (HPA: tissue IHC). | The selected field may lack the named cell population; tissue-wide labels alone do not identify positive cells (HPA: tissue IHC; general IHC practice). | Locate the relevant cells on a counterstained section before interpreting a negative result; use HPA's named cell populations to select the field (HPA: tissue IHC; general IHC practice). |
| Expected cells are present but show no chromogenic signal (HPA: tissue IHC). | Retrieval or detection may have failed, or the specimen may differ from HPA's uncertain reference pattern (general IHC practice; HPA: tissue IHC reliability). | Check a positive control and detection reagents; optimize retrieval and antibody concentration as general IHC variables, without assuming MTMR3-specific fixation sensitivity (general IHC practice). |
| Signal is diffuse throughout the section (general IHC practice). | Excess antibody, insufficient washing or prolonged chromogen development can raise background (general IHC practice). | Compare with a reagent control, then adjust concentration, washes or development time one variable at a time (general IHC practice). |
| Undetected reference cells stain strongly, such as adipocytes or cholangiocytes (HPA: tissue IHC). | Cross-reactivity or endogenous detection activity is possible; the HPA observations alone cannot identify which caused the signal (HPA: tissue IHC; general IHC practice). | Inspect reagent controls and the spatial pattern, then compare with an independent specificity control before assigning the signal to MTMR3 (general IHC practice). |
| Only a sharp membrane outline is visible (UniProt Q13615: topology; HPA: tissue IHC). | UniProt allows membrane association, but HPA describes a cytoplasmic tissue pattern; an outline alone is inconclusive (UniProt Q13615: subcellular location and topology; HPA: tissue IHC). | Recheck cellular boundaries and background, then compare the named positive cells with controls before scoring the outline as MTMR3 (HPA: tissue IHC; general IHC practice). |
| Nuclear signal dominates a chromogenic tissue section (HPA: tissue IHC). | That differs from HPA's cytoplasmic tissue description; HPA's uncertain nucleoplasmic observation comes from ICC-IF (HPA: tissue IHC and subcellular ICC-IF). | Assess counterstain overlap and reagent controls, and record the nuclear pattern as discordant with tissue IHC pending specificity assessment (general IHC practice; HPA: tissue IHC). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Use the catalog antibody’s paraffin-section workflow as the starting point, then assess MTMR3 staining by cell type and compartment.
Anti-MTMR3 A05937-1 has IHC images of human paraffin breast and prostate cancer sections and IF images of SIHA cells and human paraffin breast cancer sections (catalog image captions).
A05937-1 is the only SKU shown and is listed for human IHC and IF/ICC (catalog applications and reactivity). Its IHC captions show paraffin human breast and prostate cancer sections; its IF captions show SIHA cells and paraffin human breast cancer tissue (catalog image captions).
Which to pick: Choose A05937-1 for human paraffin-section IHC: its own IHC captions document EDTA pH 8.0 retrieval and 2 μg/ml primary antibody in breast and prostate cancer sections (catalog IHC captions). The same SKU is listed for IF/ICC and has IF images of SIHA cells and human paraffin breast cancer tissue; no cross-species choice is supported by the Human-only reactivity listing (catalog applications, reactivity and IF captions). Fixative and clonality are unreported (catalog IHC captions; catalog clone field).