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PTP4A2 / Protein tyrosine phosphatase type IVA 2 · IHC design guide
Design Immunohistochemistry for PTP4A2
Plan PTP4A2 paraffin-section IHC using the expected cytoplasmic tissue pattern and high staining in colon glandular cells (HPA tissue IHC). Interpret staining with the reported multi-gene targeting caution (HPA tissue IHC) and four isoforms in mind (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PTP4A2 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 in colon glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9740) | |
| Positive control | Appendix+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 consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining may reflect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Overexpressed in prostate tumor tissue (UniProt) | |
| Isoform / epitope | Four isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended PTP4A2 IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet: PB9740) is paired with a published nasopharyngeal carcinoma tissue protocol (PMC5649923).
| Sample | Paraffin-embedded Human Prostatic Cancer tissue; fixative not specified (datasheet PB9740) |
| Fixation | Image fixative and duration unreported (datasheet PB9740); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet PB9740) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9740) |
| Primary antibody | Rabbit anti-PTP4A2, 0.5-1μg/ml (datasheet PB9740) |
| Primary incubation | Overnight at 4 °C (datasheet PB9740) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9740) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PTP4A2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: PB9740); the published protocol also used citrate at pH 6.0 (PMC5649923).
Section 2
What Is the Expected PTP4A2 Staining Pattern?
PTP4A2 is reported in the cytoplasm, early endosome and cell membrane, with no transmembrane segment (UniProt Q12974: subcellular location and topology). In paraffin-section IHC, expect mainly cytoplasmic staining in glandular epithelium and kidney tubules (HPA: cytoplasmic expression in most tissues; High in glandular cells and kidney tubule cells). Treat this as a provisional pattern: HPA rates tissue IHC reliability Uncertain and cautions that the antibodies target protein from more than one gene (HPA: reliability).
| Cytoplasmic signal in appendix, colon or rectum glandular cells, or kidney tubule cells, with preserved tissue detail (HPA: High in these cells). | This matches the reported tissue and compartment pattern (HPA: cytoplasmic expression; High in these cells). It supports a plausible positive result, but staining alone cannot establish PTP4A2 specificity because HPA rates the IHC evidence Uncertain and flags detection of protein from more than one gene (HPA: reliability). |
| Predominantly nuclear staining, especially where cytoplasm remains clear. | A nuclear-only pattern conflicts with the supplied locations: cytoplasm, early endosome and cell membrane (UniProt Q12974: subcellular location; HPA: cytoplasmic tissue expression). Investigate artefact or nonspecific signal before scoring those cells as positive; weak membrane-associated or punctate staining cannot be dismissed solely from compartment because UniProt also reports membrane and endosomal locations (UniProt Q12974: subcellular location). |
| Strong staining in adipocytes, soft-tissue fibroblasts or skeletal-muscle myocytes. | These cell types were Not detected in the supplied HPA tissue IHC observations (HPA: adipose tissue, soft tissue and skeletal muscle). Check cross-reactivity and endogenous chromogen-producing activity as general IHC possibilities. Skeletal muscle needs special caution: UniProt reports high tissue expression there, while HPA reports myocytes Not detected; the sources do not resolve that difference (UniProt Q12974: tissue specificity; HPA: skeletal muscle). |
| Diffuse colour over epithelium, stroma and empty spaces, without cell boundaries or a clear cytoplasmic distribution. | That distribution does not resemble HPA's cellular cytoplasmic pattern (HPA: tissue IHC profile). Treat it as background until controls separate staining from chromogen deposition or endogenous detection activity; assessing tissue-free regions and a primary-antibody omission control is general IHC practice. |
| No signal in well-preserved appendix or colon glandular cells, or kidney tubules. | A negative result conflicts with the supplied High observations in those cells (HPA: appendix, colon and kidney). First assess the detection run and tissue identity using controls, then review the antibody's documented IHC-P conditions if available (general IHC practice). HPA's Uncertain reliability means a single negative section does not settle true absence or antibody performance (HPA: reliability). |
💡Expected PTP4A2 appearanceCall a section provisionally positive when glandular epithelial cells or kidney tubule cells show clear, mainly cytoplasmic signal at sites reported High by HPA; nuclear-only colour or uniform background is suspect, and specificity remains uncertain (HPA: tissue IHC profile and reliability; UniProt Q12974: subcellular location).
| Tissue and cell selection (HPA: tissue IHC profile). | Appendix, colon and rectum glandular cells and kidney tubule cells were High; adipocytes and soft-tissue fibroblasts were Not detected (HPA: named tissues and cells). Compare the named cell types within intact tissue rather than assigning one expected intensity to every cell (general IHC interpretation practice). |
| Compartment interpretation (UniProt Q12974: subcellular location and topology). | HPA describes mainly cytoplasmic tissue expression (HPA: tissue IHC profile). UniProt also places PTP4A2 at the cell membrane and early endosome and lists no transmembrane segment (UniProt Q12974: subcellular location and topology). The supplied HPA IHC data do not verify a separate membranous or punctate pattern. |
| Antibody evidence (HPA: antibody validation and tissue reliability). | Both listed antibodies, HPA003281 and CAB011204, have IHC status Uncertain; neither has an Enhanced IHC designation in the supplied record (HPA: antibodies). HPA also cautions that the tissue-staining antibodies target protein from more than one gene, so matching morphology is supportive but not proof of PTP4A2 identity (HPA: reliability). |
| Isoforms and epitope coverage (UniProt Q12974: isoforms and processing). | UniProt lists four isoforms and a chain spanning residues 1–164, with a modified cysteine at 164 (UniProt Q12974: isoforms, processing and modified residues). No antibody epitope is supplied, so the fraction of isoforms detected and any effect of that modification on staining are unknown; do not infer either from signal intensity. |
| IF/ICC question: Is a cellular fluorescence pattern established here? | No: HPA supplies no ICC-IF images or main subcellular location for this record (HPA: subcellular record). UniProt lists cytoplasm, early endosome and cell membrane as locations, but these do not validate an IF pattern for a particular antibody (UniProt Q12974: subcellular location; HPA: subcellular record). |
| Situation | Likely cause | Next action |
|---|---|---|
| High HPA reference cells appear blank (HPA: appendix, colon or kidney). | The detection run, tissue selection or antibody conditions may have failed; absence is difficult to interpret with Uncertain IHC evidence (HPA: tissue profile and reliability; general IHC practice). | Confirm the named cells on the section, check a working positive control and review the antibody's documented IHC-P detection and retrieval conditions if available (general IHC practice). Avoid declaring biological absence from one section. |
| Most signal is nuclear despite clear cytoplasm. | The pattern conflicts with the supplied cytoplasmic tissue profile and UniProt locations (HPA: tissue IHC profile; UniProt Q12974: subcellular location). Nonspecific staining is a possible general IHC explanation. | Compare the primary-antibody omission control and cell morphology; repeat evaluation on a named HPA High tissue before assigning a positive score (HPA: tissue IHC profile; general IHC practice). |
| Adipocytes or soft-tissue fibroblasts stain as strongly as nearby epithelium. | HPA records those specific cell types as Not detected (HPA: adipose tissue and soft tissue). Cross-reactivity or endogenous detection activity is possible in chromogenic IHC (general IHC practice). | Inspect control sections for endogenous colour, use the appropriate enzyme-blocking step for the detection chemistry, and score named cell types separately (general IHC practice). |
| Colour is diffuse across tissue and blank spaces. | The distribution lacks the cellular cytoplasmic pattern reported by HPA (HPA: tissue IHC profile). General IHC causes include excess background from detection reagents or inadequate washing. | Check the primary-antibody omission control, reagent-only areas and washes; interpret only signal that resolves to intact cells (general IHC practice). |
| Skeletal-muscle myocytes are negative, but another source suggests high expression. | The sources differ: UniProt reports high skeletal-muscle expression, whereas HPA reports myocytes Not detected by tissue IHC (UniProt Q12974: tissue specificity; HPA: skeletal muscle). The supplied record cannot explain the discrepancy. | Document the discrepancy and use HPA High glandular cells or kidney tubules to assess this IHC run; do not force skeletal muscle into a positive-control role (HPA: tissue IHC profile; general IHC practice). |
| A positive section shows membrane-edge or punctate signal alongside cytoplasm. | Membrane and early-endosome localization are compatible with UniProt, but HPA's supplied tissue profile reports cytoplasmic expression without validating those finer patterns (UniProt Q12974: subcellular location; HPA: tissue IHC profile). | Record the compartment separately from the main cytoplasmic result, compare controls, and avoid calling the fine pattern PTP4A2-specific without further antibody validation (HPA: Uncertain IHC reliability; general IHC practice). |
Experiment evidence
IHC & IF images showing PTP4A2 staining
PB9740 · IHCIHC analysis of PTP4A2 using anti-PTP4A2 antibody (PB9740). PTP4A2 was detected in paraffin-embedded section of Human Prostatic Cancer Tissu
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000184007-PTP4A2/tissue/cerebral+cortex#imid_3197200
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000184007-PTP4A2/tissue/colon#imid_3197126
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000184007-PTP4A2/tissue/liver#imid_3197119
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000184007-PTP4A2/tissue/kidney#imid_3197120
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PTP4A2 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the colon slide, compare glandular staining with adjacent cells that show no specific signal, without assuming every nonglandular cell is negative.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PTP4A2; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a concentration-matched nonimmune rabbit IgG isotype control (hero caption: rabbit primary antibody), plus PTP4A2-knockout tissue or cells as a biological negative. Quench endogenous peroxidase and check for endogenous biotin when using the caption’s biotin-based DAB detection on colon (hero caption: biotinylated secondary and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9740 paraffin-section caption does not state a fixative (hero caption: fixative not stated). The reported IHC conditions use heat retrieval in citrate buffer at pH 6 for 20 minutes; whether retrieval is essential is unreported (hero caption: citrate retrieval). No matched evidence establishes that frozen sections or IF are easier; in colon, assess endogenous peroxidase and biotin as possible sources of background with the reported detection method (hero caption: biotin-based DAB detection).
HPA tissue IHC evidence for PTP4A2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTP4A2 IHC Tips
Troubleshoot PTP4A2 chromogenic IHC by checking retrieval, tissue processing, cellular localisation, controls, and scoring against the documented paraffin-section workflow.
What retrieval should I try first for weak PTP4A2 staining in paraffin sections?
Begin with heat-mediated antigen retrieval in citrate buffer at pH 6 for 20 minutes on paraffin sections (datasheet PB9740). Keep the antibody at the documented 1 μg/mL overnight at 4°C while evaluating retrieval, so primary concentration does not complicate the comparison (datasheet PB9740; standard IHC practice). Include a reference section with colon glandular cells and a no-primary control, then compare cellular signal, tissue morphology, and background (HPA: high in colon glandular cells; standard IHC practice). If staining remains weak, test longer heating or another retrieval buffer on matched sections and check for tissue damage before adopting the change (standard IHC practice).
How should I assess fixation when the PTP4A2 paraffin image gives no fixative?
The PB9740 paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown from the supplied evidence (datasheet PB9740). Use a validated tissue-processing workflow, keep fixation conditions consistent across comparison sections, and record the time before fixation (standard IHC practice). To investigate a fixation effect, compare paired sections that differ in fixation while holding retrieval, primary antibody concentration, and detection conditions constant (standard IHC practice). Check cellular preservation and the expected cytoplasmic pattern before attributing patchy staining to expression, since uneven processing can complicate interpretation (HPA: cytoplasmic expression in most tissues; standard IHC practice).
Where should PTP4A2 staining appear, and when is its location suspicious?
PTP4A2 is reported at the cell membrane, early endosome, and in cytoplasm, and has no transmembrane segment (UniProt Q12974 topology and subcellular location). In chromogenic sections, assess cytoplasmic staining and possible peripheral or punctate signal within identifiable cells, using morphology to avoid counting extracellular pigment (HPA: cytoplasmic expression in most tissues; UniProt Q12974 location; standard IHC practice). Colon glandular cells and kidney tubule cells provide documented staining references, although their intensity need not match another specimen (HPA: high in colon glandular cells and kidney tubule cells; standard IHC practice). Predominantly nuclear staining calls for control review before assigning it to PTP4A2 (UniProt Q12974 location; standard IHC practice).
Can paraffin IHC identify a particular PTP4A2 isoform or terminal epitope?
PTP4A2 has 4 annotated isoforms and a phosphatase domain spanning residues 5–158, but the supplied PB9740 caption does not map its epitope (UniProt Q12974; datasheet PB9740). Report staining as PTP4A2 immunoreactivity rather than assigning an isoform without independent, isoform-specific validation (UniProt Q12974 isoforms; standard IHC practice). The annotated mature chain ends at residue 164, with a cysteine methyl ester at that position; this annotation alone cannot establish how fixation or retrieval affects antibody binding (UniProt Q12974 processing and modified residues; standard IHC practice). For an isoform claim, compare serial sections using an epitope-mapped reagent and assess concordant cellular staining (standard IHC practice).
How can I check a PTP4A2 IHC pattern by multiplex IF?
Use a separately validated IF antibody and multiplex it with a marker for the cell type being assessed, such as glandular cells when checking a colon pattern (HPA: high in colon glandular cells; standard IF practice). Choose spectrally separated fluorophores after inspecting tissue autofluorescence and include single-stain controls for bleed-through (standard IF practice). Because PTP4A2 is reported in cytoplasm, early endosomes, and at the cell membrane without a transmembrane segment, select permeabilisation according to the mapped epitope’s accessibility and verify that it preserves morphology (UniProt Q12974 topology and location; standard IF practice). No subcellular ICC/IF images are supplied here to validate a specific IF pattern (HPA subcellular record).
How do I troubleshoot diffuse brown staining with the documented PTP4A2 workflow?
The PB9740 image used 10% goat serum blocking, 1 μg/mL primary antibody, a biotinylated secondary, streptavidin–biotin detection, and DAB (datasheet PB9740). Check a no-primary section and compare staining inside intact cells with staining in damaged tissue or section edges (standard IHC practice). Include an endogenous peroxidase block and, where the biotin-based system gives persistent background, evaluate endogenous biotin controls as general chromogenic IHC measures (standard IHC practice). If background remains high, optimize blocking and washes before changing the documented primary concentration, and review whether counterstain obscures weak cytoplasmic signal (datasheet PB9740; standard IHC practice).
How should I quantify PTP4A2 staining across heterogeneous paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and tissue region before scoring, since the reported tissue pattern is mainly cytoplasmic and varies by cell type (HPA: cytoplasmic expression in most tissues; standard IHC practice). Record the percentage of positive target cells and staining intensity, then calculate an H-score using a consistent threshold and observer rule (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of viable annotated tissue and normalize cell-based measures to the number of eligible cells (standard IHC practice). Analyze matched processing batches and exclude necrotic or folded areas so technical differences do not become apparent expression differences (standard IHC practice).
What separates convincing PTP4A2 staining from chromogenic artefact?
Convincing staining should occupy identifiable cells in a plausible cytoplasmic or peripheral pattern and agree with a reference section and no-primary control (UniProt Q12974 location; HPA: cytoplasmic expression in most tissues; standard IHC practice). Compare the cell type explicitly: colon glandular cells are reported as high, while adipocytes are reported as not detected (HPA: colon glandular cells high; HPA: adipocytes not detected). Treat staining confined to section edges, necrotic regions, or unexpected nuclei cautiously, and check endogenous enzyme contribution with the appropriate detection controls (UniProt Q12974 location; standard IHC practice). HPA assigns its tissue staining uncertain reliability and cautions that the antibody may detect proteins from more than one gene, so confirm consequential findings independently (HPA: uncertain reliability and cross-gene caution).
Boster reagents
Best PTP4A2 / Protein tyrosine phosphatase type IVA 2 IHC Antibodies
PB9740 has real IHC data from a human prostatic cancer paraffin section (PB9740 image caption). Human and rat are listed as reactive species (catalog: PB9740 reactivity).
Real IHC data
Anti-PTP4A2 Antibody ®
Cat # PB9740
PB9740 is the only SKU that will render; its IHC figure shows staining in a human prostatic cancer paraffin section (PB9740 image caption). Its listed applications include IHC, while no IF figure or IF/ICC application is provided (catalog: PB9740 applications and IF images).
Which to pick: Choose PB9740 for paraffin-section tissue IHC based on its own human tissue figure; the fixative is unreported (PB9740 image caption). For IF/ICC, PB9740 has no listed application or figure, and its clonality is unspecified (catalog: PB9740 applications, IF images, clone). For cross-species planning, PB9740 lists human and rat reactivity, but its shown IHC evidence covers human tissue only (catalog: PB9740 reactivity; PB9740 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q12974 (TP4A2_HUMAN, Protein tyrosine phosphatase type IVA 2).
- Human Protein Atlas. PTP4A2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. PTP4A2 subcellular location (ICC-IF): Highest expression in RT-4: 391.9 nTPM.
- Human Protein Atlas. PTP4A2 antibody validation summary (2 antibodies).
- Over-expression of protein tyrosine phosphatase 4A2 correlates with tumor progression and poor prognosis in nasopharyngeal carcinoma. Oncotarget 2017 — PMC5649923.
- PubMed PMID:8529999 — UniProt-cited evidence.
- PubMed PMID:9018080 — UniProt-cited evidence.
- PubMed PMID:8661118 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PTP4A2 (Protein tyrosine phosphatase type IVA 2). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9740). Validated IHC antibody: PB9740 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human Prostatic Cancer tissue; fixative not specified (datasheet PB9740); 2. Fixation: Image fixative and duration unreported (datasheet PB9740); 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: Citrate pH 6 , 20 min (datasheet PB9740); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9740); 8. Primary antibody: Rabbit anti-PTP4A2, 0.5-1μg/ml (datasheet PB9740); 9. Primary incubation: Overnight at 4 °C (datasheet PB9740). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining may reflect protein from more than one gene (HPA tissue IHC); Isoform / epitope: Four isoforms; check epitope coverage across variants (UniProt); Regulation: Overexpressed in prostate tumor tissue (UniProt). Evidence: UniProt Q12974, HPA tissue IHC · Uncertain, HPA cell-line RNA · Highest expression in RT-4: 391.9 nTPM, Datasheet PB9740, PMC5649923. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PTP4A2/immunohistochemistry-ihc-guide.