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- Table of Contents
Plan chromogenic STMN1 IHC in paraffin sections around its cytoplasmic tissue pattern and cell-specific staining (HPA tissue IHC). A catalog antibody has a documented 1:50 IHC dilution; score stained cell populations and include a primary-omission control (datasheet M01194; 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 staining across most tissues; high in germinal center cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01194) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | High staining in colon endocrine cells can skew whole-tissue scoring (HPA tissue IHC) | |
| Regulation | Higher expression in acute leukemia cells (UniProt) | |
| Isoform / epitope | 2 isoforms; chain starts at residue 2; epitope impact unclear (UniProt) |
The catalog antibody’s IHC protocol uses EDTA pH 8.0 heat retrieval (datasheet M01194). Four published STMN1 IHC protocols provide tissue-specific examples (PMC9944248; PMC5564402; PMC4317896; PMC6129035).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet M01194) |
| Fixation | Image fixative and duration unreported (datasheet M01194); 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 M01194); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01194) |
| Primary antibody | Rabbit monoclonal (clone DDA-19) anti-STMN1, 1:50 (datasheet M01194) |
| Primary incubation | Overnight at 4 °C (datasheet M01194) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M01194) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | STMN1-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, including brain and lymphoid tissues. No signal in the no-primary control. |
STMN1 should appear predominantly in the cytoplasm, consistent with its cytoplasmic and cytoskeletal location and lack of a transmembrane segment (UniProt P16949). Expect strong staining in selected cells, including bone marrow hematopoietic cells, lymph node germinal center cells and intestinal endocrine cells (HPA: High). HPA reports cytoplasmic staining in most tissues, with Enhanced tissue IHC reliability and medium consistency between antibody staining and RNA expression (HPA).
| Strong cytoplasmic staining in bone marrow hematopoietic cells or lymph node germinal center cells (HPA: High). | This fits the reported cell-level IHC pattern and provides a positive comparison for the run (HPA: High in both populations). Score the specified cells and their cytoplasm; a stained tissue section does not imply that every cell type in it expresses STMN1 (HPA: cell-level tissue IHC). |
| Predominantly nuclear or membrane-like staining, with little cytoplasmic signal. | This conflicts with the expected cytoplasmic pattern (HPA: tissue IHC; UniProt P16949: cytoplasm, cytoskeleton). STMN1 has no transmembrane segment (UniProt P16949); investigate nonspecific staining, detection artefact or misleading morphology before calling the pattern positive (general IHC practice). |
| Strong staining in adipocytes or heart muscle cardiomyocytes (HPA: Not detected). | The result conflicts with HPA's cell-specific observations and may reflect cross-reactivity or endogenous detection activity (HPA: Not detected; general IHC practice). Compare neighboring cell populations and controls before interpreting it. HPA's finding applies to the named cells, not every cell in adipose tissue or heart muscle (HPA: cell-level tissue IHC). |
| Diffuse chromogen across tissue, stroma and cell boundaries, obscuring the cytoplasm. | This cannot establish the expected compartment or cell type (HPA: cytoplasmic tissue profile). Excess antibody, incomplete blocking, endogenous detection activity or uneven reagent coverage are possible general IHC causes. Judge specificity against a positive tissue section and an appropriate negative detection control (general IHC practice). |
| No cytoplasmic signal in bone marrow hematopoietic cells despite an otherwise readable section (HPA: High). | Treat this as a failed or uncertain positive comparison before scoring weak experimental samples. Check antibody and detection performance, section handling and the run's retrieval conditions (general IHC practice). HPA's High call establishes an expected cell-level observation, not a guarantee for every specimen or staining run (HPA: tissue IHC). |
| Cell type and tissue context (HPA: tissue IHC). | HPA reports High staining in marrow hematopoietic cells, lymph node germinal center cells, placental cytotrophoblasts and intestinal endocrine cells; it reports adipocytes and cardiomyocytes as Not detected. Compare named cell populations rather than assigning one intensity to an entire tissue (HPA: cell-level tissue IHC). |
| Antibody-level IHC evidence (HPA: HPA068438 and CAB010107). | Both listed antibodies have Enhanced IHC validation (HPA: antibody records). The tissue profile also has Enhanced reliability, described as medium consistency with RNA data (HPA: tissue IHC). These ratings support the reported pattern but do not validate an unlisted catalog antibody (HPA: listed antibody records). |
| Isoforms and modifications (UniProt P16949). | UniProt lists two isoforms and multiple modified residues, including phosphoserines (UniProt P16949). The supplied records give no epitope map or isoform coverage for the catalog antibody, so they cannot establish whether a particular IHC stain recognizes both isoforms or responds differently to phosphorylation. |
| What should IF/ICC show? (HPA: subcellular ICC-IF). | HPA reports an approved cytosolic location and lists ICC-IF images for A-431, U-251MG, U2OS, Rh30 and THP-1 (HPA: subcellular). This supports cytosolic localisation as a comparison, while the present section addresses chromogenic IHC in paraffin sections (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive marrow or germinal center cells lack cytoplasmic stain (HPA: High). | The run may have insufficient antibody or detection signal, or unsuitable retrieval conditions; these are general IHC possibilities, not documented STMN1-specific sensitivities. | Review the positive comparison and detection controls, then check antibody use and retrieval against the available IHC protocol (general IHC practice). Do not infer a target-specific fixation effect from this result. |
| Adipocytes or cardiomyocytes stain strongly (HPA: Not detected). | Cross-reactivity, endogenous detection activity or misplaced chromogen are possible explanations (general IHC practice). HPA's negative observation is specific to those named cells. | Inspect cell boundaries and run appropriate negative detection controls; confirm that the positive signal belongs to the scored cell type (general IHC practice). |
| The section has diffuse brown background or poor cell contrast. | Incomplete blocking, excess antibody, endogenous detection activity or uneven reagent coverage may obscure the expected cytoplasmic pattern (general IHC practice; HPA: cytoplasmic profile). | Check blocking, reagent application and counterstain, then compare with negative detection and known-positive tissue controls (general IHC practice). |
| Nuclei dominate the stain while cytoplasm is weak. | The observed compartment conflicts with cytoplasmic localisation (UniProt P16949; HPA: tissue IHC). Nonspecific staining or interpretation of dense chromogen may be involved (general IHC practice). | Reassess localisation at higher magnification and compare with a positive control showing discernible cytoplasm (general IHC practice; HPA: cytoplasmic profile). |
| A whole tissue is called negative because its named high-staining population was missed. | HPA intensities refer to defined cells, such as intestinal endocrine cells or placental cytotrophoblasts (HPA: High), rather than a uniform tissue-wide stain. | Identify the relevant cells using morphology and counterstain, then score their cytoplasm separately from surrounding cells (general IHC practice; HPA: cell-level tissue IHC). |
| A weak result is attributed to fixation or phosphorylation alone. | The supplied sources report modified residues and tissue patterns but provide no target-specific fixation sensitivity or antibody epitope evidence (UniProt P16949; HPA: tissue IHC). | Record the result as unresolved, check run controls and documented assay conditions, and avoid assigning a STMN1-specific mechanism without supporting evidence (general IHC practice). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
Troubleshoot STMN1 staining in paraffin sections by checking retrieval, cell type, cytoplasmic localisation and matched controls before interpreting signal intensity.
Catalog antibodies have IHC images from paraffin-embedded human, mouse and rat tissues; PB9560 also has an MCF7 ICC/IF image (catalog image captions).
M01194 has IHC images from paraffin-embedded human colon and colon cancer (M01194 image captions); PB9560 has IHC images from human placenta and intestinal cancer and mouse and rat brain, plus MCF7 ICC/IF imagery (PB9560 image captions). A01194S25 has a phospho-Ser24 IHC image from paraffin-embedded human breast carcinoma with phosphopeptide blocking (A01194S25 image caption).
Which to pick: For tissue IHC, choose M01194 when a rabbit monoclonal is preferred (M01194 catalog entry; M01194 image captions), or A01194S25 when the question concerns phospho-Ser24 (A01194S25 catalog entry; A01194S25 image caption). For IF/ICC, PB9560 has an MCF7 image at 2 μg/mL (PB9560 IF image caption). PB9560 has IHC images from human, mouse and rat tissues (PB9560 image captions); the paraffin-section captions do not report a fixative (catalog image captions).