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PTPN6 / Tyrosine-protein phosphatase non-receptor type 6 · IHC design guide
Design Immunohistochemistry for PTPN6
Plan chromogenic IHC for PTPN6 in paraffin sections using its observed cytoplasmic immune-tissue pattern (HPA tissue IHC). Use immune-rich tissue as a positive reference and score immune cells separately from other cell populations (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PTPN6 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 in immune tissues; nuclear location annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | Distinct cytoplasmic staining in immune-tissue cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00938-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Immune-cell staining may confound tumor-cell scoring (HPA tissue IHC) | |
| Regulation | Expression enriched in immune tissues (HPA tissue IHC) | |
| Isoform / epitope | 4 isoforms; antibody epitope coverage is unspecified (UniProt) |
Section 1
Recommended PTPN6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet M00938-2) is accompanied by one published FFPE DLBCL protocol (PMC4036221).
| Sample | Paraffin-embedded human laryngeal squamous cell carcinomas tissue; fixative not specified (datasheet M00938-2) |
| Fixation | Image fixative and duration unreported (datasheet M00938-2); 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 M00938-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00938-2) |
| Primary antibody | Mouse monoclonal (clone 8H11B10) anti-PTPN6, 2 μg/ml (datasheet M00938-2) |
| Primary incubation | Overnight at 4 °C (datasheet M00938-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00938-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PTPN6-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: Distinct cytoplasmic expression in immune tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet M00938-2); the published protocol used citrate pH 6.1 with a different antibody (PMC4036221).
Section 2
What Is the Expected PTPN6 Staining Pattern?
PTPN6 is a soluble protein found in the cytoplasm and nucleus, with no transmembrane segment (UniProt P29350 topology and subcellular location). In paraffin-section IHC, expect a chiefly cytoplasmic pattern in immune cells, including lung macrophages and lymphoid cells (HPA: tissue IHC). HPA rates its tissue profile Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).
| Distinct cytoplasmic staining in lung macrophages, lymph-node germinal center cells, or splenic white-pulp cells. | This matches the reported cell and compartment pattern (HPA: High in these cell populations; distinct cytoplasmic expression in immune tissues). Judge the named cells against neighboring cells, rather than calling the entire tissue uniformly positive. HPA's Enhanced rating supports the pattern but carries a medium RNA–protein consistency caveat (HPA: tissue IHC reliability). |
| A sharp cell-surface outline dominates, with little intracellular staining. | A membrane-only result does not fit a protein with no transmembrane segment or the reported cytoplasmic tissue pattern (UniProt P29350 topology; HPA: tissue IHC). Review whether the outline persists in a detection control before interpreting it as PTPN6 (general IHC practice). Nuclear staining alone needs separate review: PTPN6 can enter the nucleus, and HPA reports nuclear ICC-IF signal (UniProt P29350 subcellular location; HPA: ICC-IF). |
| Strong, widespread staining appears in bronchial respiratory epithelium or adrenal glandular cells. | Those particular cell populations are reported as Not detected (HPA: bronchus and adrenal gland tissue IHC). Staining there warrants investigation for cross-reactivity or endogenous detection activity (general IHC practice). This is a cell-type comparison, not a claim that every cell in either organ must be negative. |
| A diffuse brown haze covers cells and surrounding tissue without clear cell boundaries. | That distribution is hard to reconcile with the distinct cytoplasmic immune-tissue pattern (HPA: tissue IHC). Treat it as possible background before scoring cells. Non-specific reagent binding, residual endogenous enzyme activity, and insufficient washing are general chromogenic IHC possibilities, not documented PTPN6-specific effects (general IHC practice). |
| No discernible staining appears in macrophages in a lung control or lymphoid cells in an appendix control. | Both are reported High in the specified cell populations (HPA: lung macrophages; appendix lymphoid tissue). An absent result raises concern about the staining run or antibody performance; it does not alone establish biological absence. Check the intended cells, section integrity, primary antibody, and detection controls (general IHC practice). |
💡Expected PTPN6 appearanceCall a result positive when distinct, chiefly cytoplasmic staining marks the expected immune cells, with strong signal possible in HPA High populations; membrane-only outlines or broad staining of HPA Not detected cell populations require review (HPA: tissue IHC; UniProt P29350 topology).
| IHC versus IF/ICC: does nuclear signal count? | Yes, in context: UniProt places PTPN6 in cytoplasm and nucleus, and HPA ICC-IF supports nucleoplasm with additional nucleoli (UniProt P29350 subcellular location; HPA: ICC-IF). HPA tissue IHC instead describes distinct cytoplasmic expression in immune tissues (HPA: tissue IHC). Assess nuclear signal by assay and cell context; the IF observation does not set an IHC protocol. |
| Cell population and tissue | The relevant cell matters more than the organ name: lung macrophages, lymph-node germinal center cells, and splenic white-pulp cells are High, whereas bone-marrow hematopoietic cells are Low (HPA: tissue IHC). HPA also lists tissue-enhanced RNA in bone marrow and lymphoid tissue; RNA enrichment does not make bone marrow a high-staining protein control (HPA: tissue IHC). |
| Isoform context | UniProt reports four isoforms and states that isoform 1 is expressed in hematopoietic cells while isoform 2 is expressed in non-hematopoietic cells (UniProt P29350 tissue specificity). This helps frame cell-type expectations, but the supplied antibody records give no epitope or isoform coverage; do not assign a stained cell to a particular isoform from this IHC pattern alone. |
| Topology and processing | PTPN6 has no transmembrane segment, signal peptide, or propeptide in the supplied record (UniProt P29350 topology and processing). These facts support scrutiny of a membrane-only pattern. They do not establish antigen-retrieval requirements or a fixation effect; target-specific fixation sensitivity is unreported in the supplied sources. |
| Antibody evidence | HPA labels tissue IHC Enhanced and lists IHC Enhanced for HPA001466, CAB004572, and CAB072845; HPA001466 also has ICC Supported status (HPA: tissue IHC and antibody validation). These statuses support using the published patterns as comparisons, but the supplied evidence provides no dilution, retrieval setting, or interchangeability claim for a separate catalog antibody. |
| Situation | Likely cause | Next action |
|---|---|---|
| A lung section shows no macrophage staining. | The expected comparator is missing: lung macrophages are High (HPA: tissue IHC). An unsuccessful run, a missed macrophage-rich area, or a detection problem remains possible (general IHC practice). | Verify the macrophages on the section, then review primary antibody application, detection reagent, and a run control (general IHC practice). Do not infer a PTPN6-specific fixation or retrieval failure from this result alone. |
| The only conspicuous IHC signal is nuclear. | PTPN6 can localize to the nucleus, and nuclear ICC-IF is supported, although HPA describes the tissue IHC profile as distinct cytoplasmic expression in immune tissues (UniProt P29350; HPA: ICC-IF and tissue IHC). | Check the stained cell types and cytoplasmic signal in an HPA High comparator, then inspect detection controls (HPA: tissue IHC; general IHC practice). Report the compartment observed without treating the IF pattern as proof of an IHC result. |
| Many cells show a crisp membrane-only rim. | That pattern conflicts with the reported lack of a transmembrane segment and the cytoplasmic tissue IHC profile (UniProt P29350 topology; HPA: tissue IHC). Non-specific staining is one possibility (general IHC practice). | Compare a control lacking primary antibody and reassess the pattern in expected immune cells (general IHC practice; HPA: tissue IHC). Keep a membrane-only result separate from a positive cytoplasmic score. |
| Bronchial respiratory epithelial cells stain strongly across the section. | HPA reports these cells as Not detected, so the result needs scrutiny for cross-reactivity or endogenous detection activity (HPA: bronchus tissue IHC; general IHC practice). | Inspect a control lacking primary antibody and compare any immune cells present with the epithelial staining (general IHC practice). Score by cell population and record the discrepancy with the HPA bronchial epithelial observation. |
| Brown color forms a diffuse haze over cells and extracellular areas. | The haze obscures the distinct cytoplasmic pattern expected in immune tissues (HPA: tissue IHC). In chromogenic IHC, background can arise from non-specific binding, endogenous enzyme activity, or incomplete washing (general IHC practice). | Review blocking, washes, detection controls, and counterstain contrast using standard IHC controls (general IHC practice). Interpret PTPN6 only where individual positive cells and their compartment remain discernible. |
| Bone marrow looks weaker than a lymphoid control. | HPA lists bone-marrow hematopoietic cells as Low despite tissue-enhanced RNA there, while appendix lymphoid tissue is High (HPA: tissue IHC and RNA specificity). | Compare like cell populations and use an HPA High population to assess whether the run can produce clear signal (HPA: tissue IHC; general IHC practice). Do not raise the expected bone-marrow protein intensity solely because its RNA is enriched. |
Experiment evidence
IHC & IF images showing PTPN6 staining
M00938-2 · IHCIHC analysis of SHP1/PTPN6 using anti-SHP1/PTPN6 antibody (M00938-2). SHP1/PTPN6 was detected in a paraffin-embedded section of human laryng
M00938-2 · IHCIHC analysis of SHP1/PTPN6 using anti-SHP1/PTPN6 antibody (M00938-2). SHP1/PTPN6 was detected in a paraffin-embedded section of human renal
M00938-2 · IFIF analysis of SHP1/PTPN6 using anti-SHP1/PTPN6 antibody (M00938-2). SHP1/PTPN6 was detected in an immunocytochemical section of A549 cells.
A00938Y536 · IHCImmunohistochemistry (IHC) analyzes of SHP-1 (phospho-Y536) polyclonal antibody in paraffin-embedded human breast carcinoma tissue at 1:100.
M00938 · IHCImmunohistochemical analysis of paraffin-embedded rat spleen, using SHP1 Antibody.
A00938-3 · IFIF analysis of SHP1/PTPN6 using anti-SHP1/PTPN6 antibody (A00938-3). SHP1/PTPN6 was detected in immunocytochemical section of A431 cells. En
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000111679-PTPN6/tissue/cerebral+cortex#imid_530434
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000111679-PTPN6/tissue/colon#imid_530344
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000111679-PTPN6/tissue/liver#imid_530353
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000111679-PTPN6/tissue/kidney#imid_530346
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001466; green = PTPN6, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001466; green = PTPN6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PTPN6 IHC & IF
🧪Run lung first: macrophages should stain strongly (HPA: High in lung macrophages); use bronchus respiratory epithelial cells as the negative tissue comparison (HPA: Not detected in bronchus respiratory epithelial cells). On the lung slide, morphologically identified nonmacrophage cells should provide contrast with macrophages, but should not be assumed uniformly negative (HPA: High in lung macrophages).
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PTPN6 in A-431, MCF-7, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched mouse IgG isotype control for the mouse catalog antibody (selected-SKU IHC caption: mouse primary antibody; standard IHC practice); use PTPN6-knockout material as a biological specificity control (standard IHC practice). Block endogenous peroxidase and inspect lung macrophages for pigment before interpreting DAB signal (selected-SKU IHC caption: peroxidase/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU IHC caption: fixative not stated). The reported tissue IHC used heat retrieval in EDTA at pH 8.0, but retrieval dependence has not been established by a comparison in the supplied evidence (selected-SKU IHC caption: EDTA pH 8.0). The supplied evidence does not establish that frozen sections or IF are easier; ICC-IF images exist for PTPN6, while lung macrophage pigment and endogenous peroxidase can complicate chromogenic scoring (HPA: ICC-IF images in A-431, MCF-7 and U2OS; standard IHC practice).
HPA tissue IHC evidence for PTPN6
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Skin | Cells in granular layer | High | Protein (IHC) | HPA → |
| Spleen | Cells in white pulp | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTPN6 IHC Tips
Troubleshoot PTPN6 staining in paraffin sections by checking retrieval, cell identity, compartment, and controls before comparing signal across samples.
Which retrieval conditions should I try first for weak PTPN6 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M00938-2). That condition was used for the catalog antibody in a paraffin-embedded human laryngeal squamous cell carcinoma section, although the caption does not report the fixative (datasheet M00938-2). If signal is weak, compare a shorter and longer heating interval on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Include a lymph node germinal center or lung macrophage control to distinguish inadequate retrieval from a sample with few expected positive cells (HPA: High in germinal center cells and macrophages).
How can I troubleshoot fixation-related loss of PTPN6 signal?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative, and no fixation comparison is supplied (datasheet M00938-2). Record the fixative, fixation duration, tissue thickness, and time to fixation for each specimen before attributing weak staining to PTPN6 biology (standard IHC practice). Compare sections processed together from a reliably positive control, such as lung containing macrophages, using the same retrieval and detection run (HPA: High in lung macrophages; datasheet M00938-2 retrieval). If that control also weakens across processing batches, investigate processing records and repeat staining before interpreting a specimen as negative (standard IHC practice).
Should PTPN6 stain nuclei, cytoplasm, or cell membranes?
Score cytoplasmic and nuclear staining separately because PTPN6 is assigned to both compartments, while tissue IHC shows distinct cytoplasmic expression in immune tissues (UniProt P29350 subcellular; HPA: tissue IHC profile). Nuclear signal is biologically plausible, but the reported neuronal nuclear translocation after angiotensin II is annotated by similarity and should not be presumed in other specimens (UniProt P29350 subcellular). PTPN6 has no transmembrane segment, so a crisp membrane-only pattern needs confirmation with another antibody or an orthogonal assay (UniProt P29350 topology; standard IHC practice). Review hematoxylin counterstain and cell boundaries before assigning mixed chromogenic signal to either compartment (standard IHC practice).
Can this antibody distinguish PTPN6 isoforms or phosphorylation states?
Do not assign an isoform or phosphorylation state from chromogenic staining without an epitope map and corresponding validation for this antibody (standard IHC practice). PTPN6 has 4 reported isoforms; isoform 1 is expressed in hematopoietic cells and isoform 2 in non-hematopoietic cells (UniProt P29350 isoforms and tissue specificity). Its two SH2 domains span residues 4–100 and 110–213, and its phosphatase domain spans 244–515 (UniProt P29350 domains). Reported modified residues include phosphorylated serine, threonine, and tyrosine sites, but total-protein IHC alone does not establish their occupancy (UniProt P29350 modified residues; standard IHC practice).
How should I adapt the PTPN6 readout for multiplex immunofluorescence?
Treat IF as a separate assay and validate the antibody and staining conditions before using its fluorescence to confirm chromogenic IHC findings (standard IF practice). Multiplex PTPN6 with a marker identifying the expected cell population, such as a macrophage marker when assessing lung macrophages (HPA: High in lung macrophages; standard IF practice). Choose fluorophores after inspecting unstained tissue for autofluorescence, and place the weaker signal in a spectrally cleaner channel (standard IF practice). Because PTPN6 is cytoplasmic and nuclear with no transmembrane segment, optimize permeabilisation for intracellular access while checking that morphology and the cell-marker signal remain interpretable (UniProt P29350 subcellular and topology; standard IF practice).
What should I check when DAB staining is widespread?
First compare the stained section with a no-primary control to assess detection-system background, and inspect whether signal follows tissue edges or damaged areas (standard IHC practice). The selected paraffin-section example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary before DAB development (datasheet M00938-2). Use a peroxidase block and titrate the primary antibody and DAB development time if diffuse color persists; these are general chromogenic IHC steps, not PTPN6-specific validation (standard IHC practice). Compare immune-cell-rich and expected low-signal compartments on the same section before calling uniform staining biological (HPA: distinct cytoplasmic immune-tissue expression; standard IHC practice).
How should I quantify PTPN6 across sections with different immune-cell content? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region and cell population before scoring, then record cytoplasmic and nuclear staining separately when both are present (UniProt P29350 subcellular; standard IHC practice). For cell-based comparison, report percent positive cells and an H-score using intensity categories 0–3; for infiltrates, report positive-cell density per mm² of analyzed tissue (standard IHC practice). Normalize within the relevant cell population or to its total cell count, since immune-cell abundance can change whole-section signal independently of staining per cell (HPA: distinct cytoplasmic expression in immune tissues; standard IHC practice). Apply identical thresholds, retrieval, DAB development, and exclusion rules across batches, with a shared positive control (standard IHC practice).
How can I distinguish true PTPN6 positivity from staining artefact?
Expect interpretable cytoplasmic staining in immune-cell-rich tissue and check whether positive cells match the section’s morphology; lung macrophages and lymph node germinal center cells are reported high-signal populations (HPA: tissue IHC profile). Nuclear staining can be plausible, but isolated membrane-only staining conflicts with the reported intracellular localization and lack of a transmembrane segment (UniProt P29350 subcellular and topology). Reject apparent positives concentrated at section edges, folds, or necrotic areas until repeat sections show the same cellular pattern (standard IHC practice). Use a no-primary control to investigate endogenous peroxidase or detection background, and compare a positive control before calling absent staining a biological negative (standard IHC practice).
Boster reagents
Best PTPN6 / Tyrosine-protein phosphatase non-receptor type 6 IHC Antibodies
The catalog includes anti-PTPN6 antibodies with IHC images from human carcinoma and rat spleen paraffin sections, plus IF/ICC images from A431 and A549 cells (catalog image captions).
Real IHC data
Anti-SHP1/PTPN6 Antibody ® (monoclonal, 8H11B10)
Cat # M00938-2
Real IHC data
Anti-SHP-1 (phospho-Y536) PTPN6 Antibody
Cat # A00938Y536
Real IHC data
Anti-SHP1 PTPN6 Rabbit Monoclonal Antibody
Cat # M00938
Real IF data
Anti-SHP1/PTPN6 Antibody ®
Cat # A00938-3
M00938-2 shows IHC in human laryngeal squamous cell carcinoma, while A00938Y536 shows phospho-Y536 IHC in human breast carcinoma (respective IHC image captions). M00938 shows IHC in rat spleen, and A00938-3 shows IF/ICC in A431 cells (respective image captions).
Which to pick: For tissue IHC, choose M00938-2: its human laryngeal carcinoma paraffin-section image reports 2 μg/ml antibody after EDTA retrieval (M00938-2 IHC image caption); the fixative is unreported (M00938-2 IHC image caption). For IF/ICC, choose A00938-3, shown in A431 cells at 5 μg/ml (A00938-3 IF image caption). For work across species, M00938 is a rabbit monoclonal listed as reactive with human, mouse and rat (catalog: M00938); its tissue IHC image shows rat spleen paraffin sections, with the fixative unreported (M00938 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P29350 (PTN6_HUMAN, Tyrosine-protein phosphatase non-receptor type 6).
- Human Protein Atlas. PTPN6 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PTPN6 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli..
- Human Protein Atlas. PTPN6 antibody validation summary (3 antibodies).
- Epigenetic mechanisms of protein tyrosine phosphatase 6 suppression in diffuse large B-cell lymphoma: implications for epigenetic therapy. Leukemia 2014 — PMC4036221.
- Pan-cancer analysis of PTPN6: prognostic significance and functional implications in tumor progression. Discover oncology 2025 — PMC12378822.
- PTPN6-EGFR Protein Complex: A Novel Target for Colon Cancer Metastasis. Journal of oncology 2022 — PMC8856819.
- Multi-omic analyses reveal PTPN6's impact on tumor immunity across various cancers. Scientific reports 2025 — PMC11958644.
- PubMed PMID:1732748 — UniProt-cited evidence.
- PubMed PMID:1652101 — UniProt-cited evidence.
- PubMed PMID:1736296 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PTPN6 (Tyrosine-protein phosphatase non-receptor type 6). Expected localisation Cytoplasmic in immune tissues; nuclear location annotated (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00938-2). Validated IHC antibody: M00938-2 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human laryngeal squamous cell carcinomas tissue; fixative not specified (datasheet M00938-2); 2. Fixation: Image fixative and duration unreported (datasheet M00938-2); 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 M00938-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00938-2); 8. Primary antibody: Mouse monoclonal (clone 8H11B10) anti-PTPN6, 2 μg/ml (datasheet M00938-2); 9. Primary incubation: Overnight at 4 °C (datasheet M00938-2). Watch for: Fixation: Keep fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Immune-cell staining may confound tumor-cell scoring (HPA tissue IHC); Isoform / epitope: 4 isoforms; antibody epitope coverage is unspecified (UniProt); Regulation: Expression enriched in immune tissues (HPA tissue IHC). Evidence: UniProt P29350, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M00938-2, PMC4036221. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PTPN6/immunohistochemistry-ihc-guide.