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PTPRO / Receptor-type tyrosine-protein phosphatase O · IHC design guide
Design Immunohistochemistry for PTPRO
Plan chromogenic PTPRO IHC in paraffin sections using selective cytoplasmic glomerular staining as a reference (HPA tissue IHC). Start within the catalog antibody’s 1:50–1:200 IHC range (datasheet) and include a no-primary control (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PTPRO 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 | Glomerular cytoplasm (HPA tissue IHC); membrane topology (UniProt) | |
| Staining pattern | Selective cytoplasmic staining in glomerular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+2 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Splice/transcript discrepancy may affect staining concordance (HPA tissue IHC) | |
| Regulation | Stimulus-dependent regulation unreported (UniProt) | |
| Isoform / epitope | 5 isoforms; extracellular vs cytoplasmic epitope matters (UniProt) |
Section 1
Recommended PTPRO IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published PTPRO immunohistochemistry protocol (PMC4496227 methods).
| Sample | Paraffin-embedded human kidney carcinoma tissue; fixative not specified (datasheet A04781) |
| Fixation | Image fixative and duration unreported (datasheet A04781); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-PTPRO, 1:50-1:200 (datasheet A04781) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PTPRO-positive staining in cells in glomeruli of kidney (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression of glomeruli. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen-retrieval specification); the published protocol used autoclave retrieval (PMC4496227 methods).
Section 2
What Is the Expected PTPRO Staining Pattern?
PTPRO is a membrane protein with an extracellular region, one transmembrane segment, and a cytoplasmic region (UniProt Q16827 topology). In tissue IHC, expect selective staining of kidney glomerular cells, reported as cytoplasmic and high (HPA tissue IHC). HPA rates the tissue staining evidence Enhanced while noting medium agreement with RNA data and a possible splice or transcript discrepancy (HPA tissue IHC).
| High staining is concentrated in cells of kidney glomeruli (HPA tissue IHC). | This matches the strongest listed tissue IHC signal and the selective glomerular profile (HPA tissue IHC). Interpret the visible cytoplasmic pattern alongside PTPRO's membrane topology; HPA's tissue image description does not resolve which side of the membrane contains the detected epitope (HPA tissue IHC; UniProt Q16827 topology). |
| Medium staining appears in Paneth cells of duodenum or small intestine (HPA tissue IHC). | This is a second reported positive cell population, so intestinal staining is not automatically off target (HPA tissue IHC). Judge its cell specificity and intensity against the listed Paneth cell pattern; the kidney glomerulus remains the stronger reference signal (HPA tissue IHC). |
| Nuclear-only staining, with no selective glomerular pattern, dominates the section (HPA tissue IHC; UniProt Q16827 topology). | Treat this as suspect because the reported tissue pattern is cytoplasmic and PTPRO is annotated at the membrane (HPA tissue IHC; UniProt Q16827 subcellular location). Check controls and detection before calling it PTPRO; nuclear color alone does not establish target localization (general IHC practice). |
| Strong color appears in adipocytes or adrenal glandular cells (HPA tissue IHC: not detected). | These listed cell populations lack detectable HPA tissue IHC staining (HPA tissue IHC). Widespread signal there raises concern for cross-reactivity or endogenous detection activity; compare a no-primary control and review blocking and detection reagents (general IHC practice). |
| Diffuse color covers glomeruli and surrounding tissue without a clear cell pattern (HPA tissue IHC: selective glomerular expression). | Diffuse background obscures the selective result needed for interpretation (HPA tissue IHC). If a known-positive kidney section has no discernible glomerular signal, assess the run's positive control before scoring the sample negative (general IHC practice). |
💡Expected PTPRO appearanceCall a section positive when glomerular cells show selective, high cytoplasmic staining (HPA tissue IHC), interpreted in light of membrane localization (UniProt Q16827 topology); widespread color in HPA-negative cells or isolated nuclear staining is suspect (HPA tissue IHC; general IHC practice).
| Tissue and cell choice (HPA tissue IHC) | Kidney glomerular cells offer the listed high positive reference; Paneth cells in duodenum and small intestine are medium references (HPA tissue IHC). Adipocytes and adrenal glandular cells are listed as not detected, so use them to examine unwanted staining within an appropriately controlled run (HPA tissue IHC; general IHC practice). |
| Compartment assignment (UniProt Q16827 topology; HPA tissue IHC) | PTPRO has extracellular residues 30–822, a transmembrane segment at 823–843, and cytoplasmic residues 844–1216 (UniProt Q16827 topology). HPA describes tissue staining as cytoplasmic and its subcellular summary as membrane; neither record identifies the catalog antibody's epitope (HPA tissue IHC; HPA subcellular). |
| Isoforms and evidence limits (UniProt Q16827; HPA tissue IHC) | UniProt lists 5 isoforms (UniProt Q16827 isoforms). HPA flags a splice or transcript discrepancy and medium consistency between antibody staining and RNA expression (HPA tissue IHC). These facts support cautious interpretation of discordant results but do not identify which isoform an IHC signal represents. |
| Antibody evidence (HPA antibodies; HPA tissue IHC) | HPA034525 is listed as a rabbit polyclonal antibody with Enhanced IHC validation (HPA antibodies). The tissue profile is also rated Enhanced, with the stated RNA agreement caveat (HPA tissue IHC). Validation supports the reported pattern; it does not make staining in an unlisted cell type a confirmed PTPRO result. |
| IF/ICC applicability? (HPA subcellular; HPA antibodies) | HPA's subcellular summary says membrane, but it provides no ICC-IF cell-line images, and the listed antibody has no ICC validation status (HPA subcellular; HPA antibodies). Use the separate IF/ICC guide for that application; the tissue IHC pattern alone cannot establish an expected fluorescent cell image. |
| Situation | Likely cause | Next action |
|---|---|---|
| No glomerular signal in a kidney positive control (HPA tissue IHC: high in glomerular cells). | The run may have failed at retrieval, primary incubation, or chromogenic detection (general IHC practice); the supplied sources give no PTPRO-specific retrieval condition. | Review the control and each assay step, then optimize retrieval and antibody use under the laboratory's IHC workflow (general IHC practice). Do not score an unknown section negative until the positive control works. |
| Glomeruli stain, but surrounding tissue is also diffusely colored (HPA tissue IHC: selective glomerular expression). | Excess background may come from nonspecific binding or detection chemistry (general IHC practice); HPA does not attribute this pattern to PTPRO. | Compare a no-primary control, inspect blocking and washes, and adjust assay conditions using the positive reference section (general IHC practice). Preserve a readable glomerular pattern when judging the result (HPA tissue IHC). |
| Adipocytes or adrenal glandular cells show strong staining (HPA tissue IHC: not detected). | The mismatch raises concern for cross-reactivity or endogenous detection activity (general IHC practice); it does not prove either cause. | Check a no-primary control and the assay's endogenous-activity block, then reassess cell-specific staining with the kidney reference (general IHC practice; HPA tissue IHC). |
| Only nuclei appear positive in kidney tissue (HPA tissue IHC: cytoplasmic glomerular profile). | Nuclear-only color conflicts with HPA's observed profile and UniProt's membrane annotation (HPA tissue IHC; UniProt Q16827 subcellular location). | Inspect counterstain and detection controls, then repeat interpretation against a section showing selective glomerular staining (general IHC practice; HPA tissue IHC). Do not assign nuclear localization from this result alone. |
| Intestinal Paneth cells stain less strongly than glomerular cells (HPA tissue IHC). | HPA reports medium Paneth cell staining and high glomerular cell staining (HPA tissue IHC). | Compare each cell population with its own reported level and keep the kidney glomerulus as the stronger positive reference (HPA tissue IHC). A weaker intestinal result alone does not show assay failure. |
| IHC and an IF/ICC image appear to disagree (HPA tissue IHC; HPA subcellular). | HPA reports a tissue IHC pattern but supplies no ICC-IF cell-line images or ICC validation for the listed antibody (HPA tissue IHC; HPA subcellular; HPA antibodies). | Evaluate the IHC section against tissue-specific controls and the separate IF/ICC evidence for that application (general IHC practice). Do not use the unvalidated IF/ICC appearance to overturn the reported glomerular IHC profile (HPA tissue IHC; HPA antibodies). |
Experiment evidence
IHC & IF images showing PTPRO staining
A04781 · IHCImmunohistochemistry (IHC) analyzes of PTPRO pAb in paraffin-embedded human kidney carcinoma tissue at 1:50,showing cytoplasm and membrane s
A04781 · IHCImmunohistochemistry (IHC) analyzes of PTPRO pAb in paraffin-embedded human stomach carcinoma tissue at 1:50,showing cytoplasm and membrane
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151490-PTPRO/tissue/cerebral+cortex#imid_8931154
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151490-PTPRO/tissue/colon#imid_8924832
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151490-PTPRO/tissue/liver#imid_8924817
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151490-PTPRO/tissue/kidney#imid_8924834
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PTPRO IHC & IF
🧪Run kidney first: cells in glomeruli should stain strongly (HPA: High in cells in glomeruli). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the kidney slide, compare glomerular staining with adjacent nonglomerular cells, which should have little or no specific signal.
Positive control tissue: Kidney (Cells in glomeruli, 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 PTPRO; derive a cell-line control from the positive tissue's cell type (Cells in glomeruli) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary (secondary-only) control, a matched rabbit polyclonal IgG isotype control, and, if available, PTPRO knockout tissue or an immunizing-peptide block (caption: PTPRO pAb and anti-rabbit secondary; standard IHC controls). Block endogenous peroxidase and check for endogenous biotin in kidney when using avidin-peroxidase detection (caption: kidney tissue and avidin-peroxidase detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval requirement are unreported in the supplied evidence; the selected A04781 paraffin-section caption also does not report a fixative (caption: fixative unreported). The evidence does not establish whether frozen sections or IF would be easier for PTPRO, so optimize retrieval and detection on control tissue before interpreting negative staining. In kidney, endogenous biotin may complicate the caption’s avidin-peroxidase method (caption: avidin-peroxidase detection; standard IHC practice).
HPA tissue IHC evidence for PTPRO
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Caution, Splice and/or transcript discrepancy exists.). 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 |
|---|---|---|---|---|
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Duodenum | Paneth cells | Medium | Protein (IHC) | HPA → |
| Small intestine | Paneth cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTPRO IHC Tips
These questions address paraffin-section chromogenic IHC for PTPRO, with one IF-specific question for planning a companion experiment (page application setting).
What retrieval should I try first if glomerular PTPRO staining is weak?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval setting). If glomerular staining is weak, confirm that sections were fully deparaffinised and that the bath reached its set temperature before changing retrieval conditions (standard IHC practice). Compare a known glomerulus-containing section processed in the same run, then adjust heating time in small increments while watching for tissue damage (standard IHC practice; HPA: high staining in glomerular cells). Record the retrieval time alongside antibody concentration and detection conditions so a gain in signal can be distinguished from increased background (standard IHC practice).
How should I troubleshoot weak staining when fixation history is uncertain?
PTPRO-specific sensitivity to fixation is unknown from the supplied evidence, and the A04781 paraffin-section caption does not report a fixative (A04781 caption: fixative not stated). Obtain the fixation record where possible, and compare sections with documented processing using the same retrieval and staining run (standard IHC practice). Check whether glomeruli retain clear morphology and whether weak signal affects the whole section or only poorly preserved regions (standard IHC practice; HPA: glomerular expression). Keep retrieval at citrate pH 6.0, 95–98 °C for 20 minutes during that comparison; vary one processing factor at a time (page retrieval setting; standard IHC practice).
Where should PTPRO signal appear in a kidney section?
Prioritise cells in glomeruli when assessing kidney sections: HPA reports high staining there and describes selective cytoplasmic glomerular expression (HPA: tissue IHC profile). PTPRO is also annotated as a membrane protein with an extracellular region at residues 30–822 and a cytoplasmic region at 844–1216 (UniProt Q16827 topology). Accordingly, examine membrane-associated and cytoplasmic staining separately, without treating either pattern alone as proof of antibody specificity (UniProt Q16827 topology; HPA: tissue IHC profile; standard IHC practice). The A04781 paraffin-section image describes cytoplasmic and membrane staining in human kidney carcinoma; use that observation as image-specific context, not a map of normal glomeruli (A04781 caption).
Could isoforms or epitope position explain different staining patterns?
PTPRO has 5 listed isoforms, so establish which forms the antibody recognises before comparing staining across studies (UniProt Q16827 isoforms; standard IHC practice). The full-length topology places residues 30–822 outside the cell and 844–1216 inside, with a membrane span at 823–843 (UniProt Q16827 topology). Ask for the immunogen sequence or mapped epitope; the supplied A04781 caption gives no epitope location, so staining cannot identify the recognised isoform (A04781 caption; standard IHC practice). The extracellular region includes multiple annotated glycosylation sites, but their effect on this antibody’s staining is untested here (UniProt Q16827 glycosylation; A04781 caption).
How would I adapt the cell-localisation question to multiplex IF?
Pair PTPRO with a validated marker of the expected glomerular cell population and inspect each channel separately before interpreting overlap (HPA: high staining in glomerular cells; standard IF practice). Choose a far-red fluorophore for a dim PTPRO channel when tissue autofluorescence is prominent, and include single-stain and no-primary controls (standard IF practice). If the antibody recognises the extracellular region, begin with minimal permeabilisation; a cytoplasmic epitope generally requires permeabilisation after fixation (UniProt Q16827 topology; standard IF practice). Confirm the antibody’s epitope and IF performance independently, because the supplied A04781 evidence documents paraffin-section chromogenic IHC rather than IF (A04781 caption).
How can I reduce diffuse staining without losing glomerular signal?
First compare the stained section with a no-primary control and inspect whether colour develops in glomeruli or broadly across the slide (standard IHC practice; HPA: selective glomerular expression). The A04781 image used PBS instead of primary antibody for its negative control and used a biotin-based, peroxidase-linked detection sequence (A04781 caption). Block endogenous peroxidase, assess endogenous biotin when using avidin-biotin detection, and titrate the primary antibody against background (standard IHC practice; A04781 caption: detection sequence). Its reported 1:50 dilution belongs to that pictured carcinoma-section experiment; optimise dilution on your own sections while keeping exposure and development conditions matched (A04781 caption; standard IHC practice).
What should I measure when comparing PTPRO staining between sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define glomeruli as the analysis region before scoring, because HPA reports selective cytoplasmic staining and high expression in glomerular cells (HPA: tissue IHC profile). For a cell-based readout, record the percentage of positive glomerular cells and an H-score using intensity categories 0–3; apply one threshold consistently (standard IHC practice). For area-based analysis, report positive signal per glomerular area in mm² and normalise comparisons to the measured glomerular area or cell count (standard IHC practice). Exclude folds, torn tissue and necrotic regions, and keep retrieval, illumination and chromogen development consistent across the batch (standard IHC practice).
How do I distinguish convincing PTPRO staining from artefact?
A convincing kidney pattern should involve glomerular cells, where HPA reports high staining, and should fit the documented cytoplasmic or membrane-associated observations (HPA: tissue IHC profile; UniProt Q16827 topology). Treat strong nuclear-only staining or widespread staining in unrelated cells as a specificity concern requiring controls, rather than assigning it to PTPRO from colour alone (UniProt Q16827 topology; standard IHC practice). Check section edges and necrotic areas for exaggerated deposition, and compare with a no-primary control to identify detection-system or endogenous-enzyme signal (standard IHC practice; A04781 caption: no-primary control). Interpret discordant samples cautiously because HPA reports only medium staining–RNA consistency and flags splice or transcript discrepancy (HPA: Enhanced reliability description).
Boster reagents
Best PTPRO / Receptor-type tyrosine-protein phosphatase O IHC Antibodies
A04781 has real IHC data from a human kidney carcinoma paraffin section (A04781 figure caption); its listed reactivity covers human, mouse and rat (catalog: A04781). No IF data are supplied (catalog: A04781).
Real IHC data
Anti-PTPRO Antibody
Cat # A04781
A04781 is the only SKU that will render (catalog: A04781). Its IHC figure shows staining in a human kidney carcinoma paraffin section at 1:50 (A04781 figure caption); the catalog lists human, mouse and rat reactivity (catalog: A04781).
Which to pick: Choose A04781 for paraffin-section IHC: it is a rabbit polyclonal antibody (catalog: A04781; A04781 figure caption) with an IHC range of 1:50–1:200 (datasheet: A04781), and its own figure shows human kidney carcinoma staining (A04781 figure caption). The fixative is unreported (A04781 figure caption). For mouse or rat IHC, A04781 has listed reactivity but no supplied species-specific IHC figure (catalog: A04781; A04781 figure caption); for IF/ICC, no SKU in this payload lists that application or supplies IF data (catalog: A04781).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16827 (PTPRO_HUMAN, Receptor-type tyrosine-protein phosphatase O).
- Human Protein Atlas. PTPRO tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PTPRO subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. PTPRO antibody validation summary (1 antibodies).
- PTPRO represses breast cancer lung metastasis by inhibiting the JAK2-YAP axis. Scientific reports 2025 — PMC11868365.
- PTPRO knockdown protects against inflammation in hemorrhage shock-induced lung injury involving the NF-κB signaling pathway. Respiratory research 2022 — PMC9335982.
- Protein tyrosine phosphatase PTPRO represses lung adenocarcinoma progression by inducing mitochondria-dependent apoptosis and restraining tumor metastasis. Cell death & disease 2024 — PMC10770368.
- PTPRO-mediated autophagy prevents hepatosteatosis and tumorigenesis. Oncotarget 2015 — PMC4496227.
- PubMed PMID:7753550 — UniProt-cited evidence.
- PubMed PMID:7665166 — UniProt-cited evidence.
- PubMed PMID:10498613 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PTPRO (Receptor-type tyrosine-protein phosphatase O). Expected localisation Glomerular cytoplasm (HPA tissue IHC); membrane topology (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04781 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in glomeruli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human kidney carcinoma tissue; fixative not specified (datasheet A04781); 2. Fixation: Image fixative and duration unreported (datasheet A04781); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-PTPRO, 1:50-1:200 (datasheet A04781); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Splice/transcript discrepancy may affect staining concordance (HPA tissue IHC); Isoform / epitope: 5 isoforms; extracellular vs cytoplasmic epitope matters (UniProt); Regulation: Stimulus-dependent regulation unreported (UniProt). Evidence: UniProt Q16827, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A04781, PMC4496227. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PTPRO/immunohistochemistry-ihc-guide.