STMN1 / Stathmin · IHC design guide

Design Immunohistochemistry for STMN1

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).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for STMN1 (IHC for STMN1): expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC), antibody M01194, validated IHC image, and IHC protocol steps
Printable STMN1 IHC protocol sheet — expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC), antibody M01194, controls and protocol steps. Open the full STMN1 IHC guide →

STMN1 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended STMN1 IHC & IF Protocols

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).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human colon tissue; fixative not specified (datasheet M01194)
FixationImage fixative and duration unreported (datasheet M01194); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet M01194); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet M01194)
Primary antibodyRabbit monoclonal (clone DDA-19) anti-STMN1, 1:50 (datasheet M01194)
Primary incubationOvernight at 4 °C (datasheet M01194)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet M01194)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultSTMN1-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.
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet M01194); the published HSCC protocol provides a citrate pH 6.0 comparison (PMC9944248).
Section 2

What Is the Expected STMN1 Staining Pattern?

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).

What am I looking at on my slide?
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).
💡Expected STMN1 appearanceCall a positive result when the specified high-expressing cells show clear cytoplasmic staining (HPA: High; HPA: cytoplasmic tissue profile); dominant nuclear or membrane-like staining, or strong staining in HPA-negative cell types, calls for investigation before scoring (UniProt P16949; HPA: Not detected).
How each factor affects the staining
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).
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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).

Sample controls for STMN1 IHC & IF

🧪Run colon first and assess its endocrine cells for strong staining (HPA: High in colon endocrine cells); use adipose tissue as the negative tissue and assess its adipocytes for absent staining (HPA: Not detected in adipocytes). On the colon slide, compare adjacent nonendocrine epithelial cells with the endocrine cells as an internal lower-signal reference, without requiring complete absence because STMN1 is broadly expressed (UniProt P16949: ubiquitous).
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show STMN1 in A-431, U-251MG, U2OS, Rh30, THP-1, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a species- and isotype-matched nonimmune primary control appropriate to the antibody’s clonality; and STMN1 knockout material as a biological specificity control (selected caption: rabbit primary antibody; standard IHC practice). For colon DAB staining, quench endogenous peroxidase and check the no-primary slide for residual background (selected caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative is unreported in the selected M01194 tissue-IHC caption; its paraffin section does not establish fixation (selected M01194 caption). The demonstrated colon procedure used heat-mediated EDTA retrieval at pH 8.0, a 1:50 primary dilution, and overnight incubation at 4°C (selected M01194 caption). ICC/IF images support cytosolic localisation in the listed cell lines, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC; in colon, endogenous peroxidase can contribute DAB background (HPA: Cytosol, approved; standard IHC practice). The selected M01194 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image M01194).

HPA tissue IHC evidence for STMN1

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced STMN1 IHC Tips

Troubleshoot STMN1 staining in paraffin sections by checking retrieval, cell type, cytoplasmic localisation and matched controls before interpreting signal intensity.

Where should I start when STMN1 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet M01194). The documented colon-section example then used 10% goat serum, primary antibody at 1:50 overnight at 4°C, and a peroxidase-based DAB readout (datasheet M01194). If staining remains weak, vary retrieval heating and cooling within your laboratory’s established workflow while keeping a known-positive section in each run (standard IHC practice). Judge any gain by clearer cytoplasmic staining in the expected cells, rather than by a general increase in brown signal (HPA: cytoplasmic expression in most tissues; standard IHC practice).
Could fixation explain weak or uneven STMN1 staining?
The selected paraffin-section caption does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet M01194). Record the fixative, time before fixation and fixation duration for each specimen, then compare sections processed together under the same retrieval and detection conditions (standard IHC practice). If signal varies across a section, inspect morphology and compare its interior with its cut edges before changing antibody concentration; uneven preservation can complicate interpretation (standard IHC practice). Use a matched known-positive section to test whether a processing change restores the expected cytoplasmic pattern, and avoid assigning the effect specifically to STMN1 without that comparison (HPA: cytoplasmic expression in most tissues; standard IHC practice).
Which staining compartment should count as an STMN1-positive result?
Score cytoplasmic staining as the expected pattern: STMN1 is assigned to the cytoplasm and cytoskeleton, and HPA reports cytoplasmic tissue staining (UniProt P16949 localisation; HPA: tissue IHC profile). Its approved subcellular location is the cytosol, and the record lists no transmembrane segment (HPA: subcellular; UniProt P16949 topology). Evaluate the relevant cells within the tissue rather than treating every brown structure as positive; for example, HPA reports high staining in colon endocrine cells (HPA: colon endocrine cells, High). Predominantly nuclear or sharp membrane-only staining merits review of controls, morphology and detection background before it is scored as STMN1 (HPA: subcellular; standard IHC practice).
Can isoforms or phosphorylation change how I interpret an STMN1 stain?
The record lists 2 STMN1 isoforms and an SLD spanning residues 4–145, but the supplied product caption does not identify the antibody’s epitope (UniProt P16949 isoforms and domains; datasheet M01194). STMN1 also has reported modified residues, including phosphoserines at positions 4, 16 and 25 (UniProt P16949 modified residues). Check the antibody’s documented immunogen or epitope before claiming that a negative section lacks a particular isoform or phosphorylation state (standard IHC interpretation). With epitope coverage unspecified here, report the result as staining obtained with this antibody and compare specimens under identical processing conditions (datasheet M01194; standard IHC practice).
How should I plan a complementary STMN1 immunofluorescence experiment?
Treat IF/ICC as a separate assay from the documented paraffin-section chromogenic example, whose staining procedure does not establish IF conditions (datasheet M01194; standard assay validation practice). Pair STMN1 with a marker for the cell population under study; a colon endocrine-cell marker is relevant when examining the HPA-reported high-staining colon population (HPA: colon endocrine cells, High; standard IF practice). Choose spectrally separated fluorophores and assess an unstained section for tissue autofluorescence before selecting the STMN1 channel (standard IF practice). Because STMN1 is cytosolic and has no transmembrane segment, establish permeabilisation for access to an intracellular epitope, using matched controls to check background and preserved cell structure (HPA: subcellular; UniProt P16949 topology; standard IF practice).
What should I change when DAB background obscures STMN1-positive cells?
First inspect a no-primary control to separate signal associated with the primary antibody from staining caused by the detection workflow (standard IHC practice). The documented colon example used 10% goat serum for blocking, 1:50 primary antibody overnight at 4°C, and peroxidase-based DAB detection (datasheet M01194). Review blocking, washes and primary-antibody concentration against that documented setup, changing one variable at a time while retaining a known-positive section (datasheet M01194; standard IHC practice). Include an endogenous-peroxidase block and check whether DAB is depositing outside intact cells; these are general chromogenic IHC controls, not evidence of an STMN1-specific background mechanism (standard IHC practice).
How can I compare STMN1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then quantify cytoplasmic signal within those cells (HPA: tissue IHC profile; standard IHC practice). An H-score combines staining intensity and the percentage of cells at each intensity; alternatively, report the percentage of positive cells or positive-cell density per mm² of evaluable tissue (standard IHC practice). Normalise cell counts to the relevant cell population, or density to intact tissue area, and apply the same threshold, imaging settings and exclusion rules across specimens (standard IHC practice). Keep colon endocrine cells separate from other colon cells when using that tissue, because HPA identifies them as a high-staining population (HPA: colon endocrine cells, High).
How do I distinguish a true STMN1 signal from artefact?
A convincing result places signal in the cytoplasm of intact cells and aligns with the population being assessed; HPA reports high staining in bone-marrow hematopoietic cells and colon endocrine cells (HPA: tissue IHC profile; HPA: bone marrow and colon, High). Treat dominant nuclear or membrane-only staining cautiously because the approved subcellular location is cytosolic and the protein has no transmembrane segment (HPA: subcellular; UniProt P16949 topology). Compare suspicious staining at section edges, in necrotic areas or across unrelated cells with the matched no-primary control (standard IHC practice). If brown signal persists without primary antibody, investigate the detection reagents and endogenous peroxidase before calling those cells STMN1-positive (standard IHC practice).
Boster reagents

Best STMN1 / Stathmin IHC Antibodies

Catalog antibodies have IHC images from paraffin-embedded human, mouse and rat tissues; PB9560 also has an MCF7 ICC/IF image (catalog image captions).

Real IHC data IHC analysis of STMN1 using anti-STMN1 antibody (M01194). STMN1 was detected in a paraffin-embedded section of human colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-STMN1 Antibody (M01194) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-Stathmin 1 STMN1 Rabbit Monoclonal Antibody
Cat # M01194
Real IHC data IHC analysis of Stathmin 1 using anti-Stathmin 1 antibody (PB9560). Stathmin 1 was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Stathmin 1 Antibody (PB9560) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-Stathmin 1/STMN1 Antibody ®
Cat # PB9560
Real IHC data Immunohistochemistry analysis of paraffin-embedded human breast carcinoma, using Stathmin 1 (Phospho-Ser24) Antibody. The picture on the right is blocked with the phospho peptide.
Anti-Phospho-Op18 (S25) STMN1 Antibody
Cat # A01194S25

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).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P16949 (STMN1_HUMAN, Stathmin).
  2. Human Protein Atlas. STMN1 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. STMN1 subcellular location (ICC-IF): Localized to the cytosol..
  4. Human Protein Atlas. STMN1 antibody validation summary (2 antibodies).
  5. Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression. Molecular and clinical oncology 2023 — PMC9944248.
  6. Stathmin1 expression is associated with aggressive phenotypes and cancer stem cell marker expression in breast cancer patients. International journal of oncology 2017 — PMC5564402.
  7. Stathmin1 regulates p27 expression, proliferation and drug resistance, resulting in poor clinical prognosis in cholangiocarcinoma. Cancer science 2014 — PMC4317896.
  8. An Immunohistochemical Study of Stathmin 1 Expression in Osteosarcoma Shows an Association with Metastases and Poor Patient Prognosis. Medical science monitor : international medical journal of experimental and clinical research 2018 — PMC6129035.
  9. PubMed PMID:2760073 — UniProt-cited evidence.
  10. PubMed PMID:2358074 — UniProt-cited evidence.
  11. PubMed PMID:1917919 — UniProt-cited evidence.