This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PPP4C / Serine/threonine-protein phosphatase 4 catalytic subunit · IHC design guide
Design Immunohistochemistry for PPP4C
Plan chromogenic PPP4C IHC in paraffin sections with the catalog antibody at 2–5 μg/mL (datasheet A06390-4). Use colon glandular cells as a high-staining reference and assess cytoplasmic staining with possible nuclear staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP4C 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, with nuclear staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Glandular cells stain strongly; cytoplasmic, sometimes nuclear (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06390-4) | |
| Positive control | Colon+4 more · see all | |
| Negative control | Bone marrow+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–307 (UniProt) |
Section 1
Recommended PPP4C IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published PPP4C tissue protocols (datasheet A06390-4; PMC11058967; PMC13263375; PMC13288592).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06390-4) |
| Fixation | Image fixative and duration unreported (datasheet A06390-4); 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 A06390-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06390-4) |
| Primary antibody | Rabbit anti-PPP4C, 2-5 μg/ml (datasheet A06390-4) |
| Primary incubation | Overnight at 4 °C (datasheet A06390-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06390-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PPP4C-positive staining in glandular cells of colon (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, additional nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A06390-4). If controls warrant adjustment, compare the reported citrate retrieval method (PMC11058967).
Section 2
What Is the Expected PPP4C Staining Pattern?
PPP4C should show mainly cytoplasmic staining in tissue IHC, with nuclear staining in some tissues (HPA tissue IHC: Supported; medium consistency with RNA). High staining is reported in colon, duodenal, gallbladder and prostate glandular cells, kidney tubule cells, lymph node germinal center cells, nasopharyngeal respiratory epithelial cells and pancreatic exocrine cells (HPA tissue IHC). PPP4C is also assigned to the nucleus and centrosome (UniProt P60510: subcellular location) and has no transmembrane segment (UniProt P60510: topology).
| Cytoplasmic staining in kidney tubule cells or colon glandular cells, with possible nuclear staining. | This fits the reported tissue pattern; both cell types have high staining, while nuclear expression occurs in several tissues (HPA tissue IHC: Supported). Judge the compartment in identified cells, since the reported level applies to a cell population within a tissue (HPA tissue IHC). |
| A sharp extracellular or luminal deposit is the dominant apparent PPP4C signal. | That distribution is difficult to reconcile with reported cytoplasmic and nuclear locations and the absence of a signal peptide or transmembrane segment (UniProt P60510: location, processing and topology). Check morphology and staining controls before calling it specific (standard IHC practice). |
| Strong staining appears chiefly in bone marrow hematopoietic cells or liver cholangiocytes. | Those particular cell populations were reported as not detected (HPA tissue IHC). Recheck cell identification and controls; discordance raises possible nonspecific antibody binding or endogenous chromogen activity, but one slide does not establish which cause applies (standard IHC practice). |
| Chromogen haze covers cells and surrounding tissue without a clear cellular pattern. | This does not resolve the reported cytoplasmic pattern (HPA tissue IHC). Inspect no-primary and detection controls for background from the detection workflow, and review blocking and washing (standard IHC practice); haze alone cannot establish PPP4C expression. |
| No convincing staining appears in kidney tubule cells or pancreatic exocrine glandular cells. | Both are reported as high-staining populations (HPA tissue IHC). A blank result prompts checks of tissue identity, retrieval, antibody dilution and detection controls (standard IHC practice). HPA tissue IHC is Supported with medium RNA agreement, so a single absent signal is not proof of biological absence (HPA tissue IHC). |
💡Expected PPP4C appearanceCall a positive result when identifiable glandular, tubular or other reported high-staining cells show clear cytoplasmic chromogen, with nuclear staining permissible in some tissues (HPA tissue IHC: Supported); dominant extracellular deposits or diffuse haze warrant artifact checks (UniProt P60510: topology; standard IHC practice).
| Cell population and compartment | PPP4C has low tissue RNA specificity, while IHC intensity differs by identified cell type (HPA tissue IHC). Compare like cells across sections; a negative report for cholangiocytes, for example, does not classify every liver cell as negative (HPA tissue IHC). |
| Antibody evidence | HPA043837 is listed as Supported for IHC and ICC, and the tissue IHC profile has medium consistency with RNA (HPA antibody record; HPA tissue IHC). Treat agreement with the reported pattern as supporting evidence, then use controls to assess an individual run (standard IHC practice). |
| Protein location and processing | Cytoplasm, nucleus and centrosome are assigned locations; no signal peptide or transmembrane segment is annotated (UniProt P60510). Extracellular or surface-only IHC is therefore an unexpected dominant pattern. The uncertain plasma membrane signal in ICC-IF should not be treated as firm IHC localization evidence (HPA subcellular ICC-IF). |
| Retrieval and chromogenic detection | Antigen retrieval conditions and peroxidase or other endogenous detection activity can affect slide interpretation (standard IHC practice). This record supplies no PPP4C-specific retrieval optimum or fixation sensitivity; assess the run with appropriate controls rather than attributing a failed stain to a documented PPP4C fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high-staining population is weak or blank. | The result conflicts with reported kidney tubule or pancreatic exocrine staining (HPA tissue IHC); a workflow failure remains possible (standard IHC practice). | Confirm the cell population and inspect a positive control, retrieval, dilution and detection steps (standard IHC practice). Do not infer a PPP4C-specific fixation effect from this comparison. |
| The entire section is brown, including areas without a defined cell pattern. | Broad background can arise from detection activity or insufficient washing (standard IHC practice); it cannot be assigned to PPP4C from appearance alone. | Compare no-primary and detection controls, check endogenous activity blocking where applicable, and review washes (standard IHC practice). Score only resolved cellular staining. |
| Signal is concentrated in luminal material or outside cells. | That location conflicts with the reported intracellular distribution and lack of a signal peptide (UniProt P60510: location and processing). | Check the same area on a counterstained section for cell boundaries and compare the no-primary control (standard IHC practice); withhold a specific call until cellular staining is resolved. |
| Bone marrow hematopoietic cells or smooth muscle cells stain strongly. | Those named cell populations were reported as not detected (HPA tissue IHC); cell misidentification, nonspecific binding or detection background are possibilities (standard IHC practice). | Verify morphology, examine the control stain and compare a reported high-staining population in the same run (HPA tissue IHC; standard IHC practice). |
| Nuclear staining appears alongside a clear cytoplasmic pattern. | Additional nuclear expression occurs in several tissues, and PPP4C is assigned to the nucleus (HPA tissue IHC; UniProt P60510). | Record cytoplasmic and nuclear staining separately in identified cells (standard IHC practice). Do not reject the section solely because nuclear signal is present. |
| Q: Does an IF/ICC image establish where IHC chromogen must appear? | A: ICC-IF reports supported nucleoplasm and cytosol, plus an uncertain plasma membrane location (HPA subcellular ICC-IF); tissue IHC reports general cytoplasmic and occasional nuclear expression (HPA tissue IHC). | Interpret the IHC section against its tissue and cell-specific IHC profile; use the ICC-IF record as compartment context, keeping its membrane assignment uncertain (HPA tissue IHC; HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing PPP4C staining
A06390-4 · IHCIHC analysis of PPP4C using anti-PPP4C antibody (A06390-4). PPP4C was detected in a paraffin-embedded section of human breast cancer tissue.
A06390-4 · IHCIHC analysis of PPP4C using anti-PPP4C antibody (A06390-4). PPP4C was detected in a paraffin-embedded section of human Invasive urothelial c
A06390-4 · IFIF analysis of PPP4C using anti-PPP4C antibody (A06390-4) and anti-Tubulin Alpha antibody (M03989-3). PPP4C was detected in immunocytochemic
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000149923-PPP4C/tissue/cerebral+cortex#imid_12130959
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000149923-PPP4C/tissue/colon#imid_13315493
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000149923-PPP4C/tissue/liver#imid_13315485
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000149923-PPP4C/tissue/kidney#imid_13315537
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA043837; green = PPP4C, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA043837; green = PPP4C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP4C IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells); use smooth muscle as the negative tissue, assessing its smooth muscle cells (HPA: Not detected in smooth muscle cells). On the colon slide, unstained nonglandular cells, if present, can indicate background, but their PPP4C-negative status is not established by the supplied HPA row (HPA: High in glandular cells).
Positive control tissue: Colon (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP4C in Hep-G2, MCF-7, U2OS, KOLF2.1J, NIH 3T3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a concentration-matched nonimmune rabbit IgG control (selected IHC caption: rabbit primary; standard IHC practice); use identically processed PPP4C-knockout material, if available, as a biological negative (standard IHC practice). Quench endogenous peroxidase and check colon sections for nonspecific DAB deposition before scoring glandular cells (selected IHC caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the A06390-4 paraffin-section caption does not state the fixative (selected IHC caption). That caption reports heat retrieval in EDTA at pH 8.0 before chromogenic detection; it does not establish that retrieval is required under other conditions (selected IHC caption). The supplied evidence does not establish whether frozen sections or IF/ICC are easier; in colon, distinguish cellular staining from any DAB deposit in gland lumens (standard IHC practice).
HPA tissue IHC evidence for PPP4C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PPP4C IHC Tips
Troubleshoot PPP4C staining in paraffin sections by checking retrieval, compartment, cell type, and controls before interpreting signal intensity.
How should I retrieve PPP4C in paraffin sections when staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06390-4). The reported breast cancer section used this retrieval before incubation with 2 μg/ml catalog antibody overnight at 4°C (datasheet A06390-4). Keep section thickness, heating, cooling, and antibody incubation consistent while comparing retrieval runs (standard IHC practice). If staining remains weak, test another retrieval buffer or heating duration on adjacent sections as a fallback, with matched controls (standard IHC practice). Judge improvement by cellular staining and preserved morphology, because PPP4C can appear in cytoplasm and nucleus (HPA: tissue IHC profile).
Could fixation explain variable PPP4C staining between paraffin blocks?
PPP4C-specific sensitivity to fixative choice or fixation duration is unknown from the supplied evidence (datasheet A06390-4: fixative not stated). The catalog image establishes staining in a paraffin-embedded breast cancer section, but its caption does not identify the fixative (datasheet A06390-4). Record the fixative and processing history for each block, then compare similarly processed sections with the same pH 8.0 EDTA retrieval (standard IHC practice; datasheet A06390-4). Use a matched positive control to distinguish a block-wide processing problem from a local staining problem (standard IHC practice). Do not assign a PPP4C-specific fixation effect from tissue staining patterns alone (HPA: tissue IHC profile).
Should PPP4C stain nuclei, cytoplasm, or centrosomes in tissue IHC?
Expect broad cytoplasmic staining with additional nuclear staining in several tissues (HPA: tissue IHC profile). PPP4C is also annotated in the nucleus, cytoplasm, and centrosome-associated cytoskeleton (UniProt P60510: subcellular location). Cell-image evidence places it mainly in the nucleoplasm and also in the cytosol; plasma-membrane localisation is uncertain (HPA: subcellular profile). Score nuclear and cytoplasmic chromogenic signal separately within the same cell type, using the counterstain to define nuclei (standard IHC practice). A diffuse membrane-only pattern warrants scrutiny against no-primary controls and tissue morphology before being called PPP4C (HPA: subcellular profile; standard IHC practice).
Could an isoform or terminal modification change PPP4C staining?
The supplied record lists 0 annotated isoforms for the 307-aa PPP4C protein, so an isoform-specific staining explanation lacks support here (UniProt P60510: isoforms and length). Its annotated terminal modifications include N-acetylalanine at residue 2 and leucine methyl ester at residue 307 (UniProt P60510: modified residues). The antibody epitope is not supplied, so neither modification can be linked to staining loss for this reagent (datasheet A06390-4: epitope not stated). PPP4C has no annotated transmembrane segment or glycosylation sites (UniProt P60510: topology and glycosylation). Compare retrieval conditions and matched controls before attributing a discrepant pattern to epitope masking (standard IHC practice).
How should I assess PPP4C localisation by multiplex IF?
Use IF as a separate validation experiment, since the selected catalog evidence describes chromogenic staining in a paraffin section (datasheet A06390-4). Pair PPP4C with a marker identifying the expected cell population, such as glandular cells in colon, and include a nuclear counterstain (HPA: high in colon glandular cells; standard IF practice). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before assigning a weak PPP4C channel (standard IF practice). Because PPP4C has no transmembrane segment and has nuclear and cytoplasmic locations, assess permeabilisation for intracellular epitope access (UniProt P60510: topology and localisation; standard IF practice). Evaluate nuclear and cytosolic signals separately, with no-primary and single-stain controls (HPA: subcellular profile; standard IF practice).
What causes diffuse brown background in PPP4C chromogenic IHC?
Check no-primary and secondary-only sections to locate background from detection reagents or endogenous tissue activity (standard IHC practice). The reported workflow used 10% goat serum blocking, a peroxidase-linked secondary, and DAB development (datasheet A06390-4). Include an endogenous peroxidase block for HRP-DAB staining, and compare blocking, washes, and development across adjacent sections (standard IHC practice). Titrate the primary around the reported 2 μg/ml condition while retaining a positive control on each run (datasheet A06390-4; standard IHC practice). Treat staining over tissue edges, folds, or damaged regions cautiously, especially when it lacks a coherent cellular pattern (standard IHC practice).
How can I quantify PPP4C without conflating cell composition and intensity? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and exclude folds, necrosis, and poorly preserved tissue before measuring chromogenic signal (standard IHC practice). Report nuclear and cytoplasmic PPP4C separately because both locations are supported, while their balance can differ across samples (HPA: tissue IHC profile; UniProt P60510: localisation). For each compartment, record percent positive cells and intensity on a 0–3 scale; an H-score spans 0–300 (standard IHC practice). Alternatively, report positive-cell density per mm² of viable tissue within the same cell class (standard IHC practice). Normalise comparisons to the number or area of eligible cells and apply identical thresholds across slides (standard IHC practice).
How do I separate credible PPP4C staining from artefact?
A credible result has cellular cytoplasmic staining, with possible nuclear staining, in an anatomically plausible population (HPA: tissue IHC profile). High staining is reported in colon glandular cells and kidney tubular cells, while bone marrow hematopoietic cells and smooth muscle cells were not detected in the listed HPA observations (HPA: tissue IHC profile). Compare suspicious membrane-only signal with the supported nucleoplasmic and cytosolic locations (HPA: subcellular profile). Reject interpretations driven by section edges, necrosis, or residual endogenous peroxidase signal after control review (standard IHC practice). HPA tissue staining has supported reliability with medium RNA–protein consistency, so a single discordant field should prompt replication (HPA: tissue IHC reliability).
Boster reagents
Best PPP4C / Serine/threonine-protein phosphatase 4 catalytic subunit IHC Antibodies
A06390-4 has human-reactive IHC and IF/ICC applications, with paraffin-section images from four human cancers and an IF image from U2OS cells (catalog applications, reactivity, and image captions).
Real IHC data
Anti-PPP4C Antibody ®
Cat # A06390-4
A06390-4 is shown in paraffin sections of human breast cancer, invasive bladder carcinoma with squamous differentiation, lung cancer, and ovarian serous adenocarcinoma (catalog IHC image captions). The same SKU has IF/ICC data in U2OS cells and is listed as human-reactive (catalog IF image caption; catalog applications and reactivity).
Which to pick: Choose A06390-4 for human paraffin-section chromogenic IHC: its own captions show EDTA retrieval at pH 8.0, 2 μg/ml primary antibody, and DAB detection; the fixative is unreported (catalog IHC image captions). For IF/ICC, A06390-4 has an image in U2OS cells using 5 μg/ml primary antibody (catalog IF image caption). No cross-species recommendation is supported because only human reactivity is listed; clonality is unreported (catalog reactivity and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P60510 (PP4C_HUMAN, Serine/threonine-protein phosphatase 4 catalytic subunit).
- Human Protein Atlas. PPP4C tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PPP4C subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane and cytosol..
- Human Protein Atlas. PPP4C antibody validation summary (1 antibodies).
- High PPP4C expression predicts poor prognosis in diffuse large B-cell lymphoma. Clinical and experimental medicine 2024 — PMC11058967.
- PPP4C-related metastasis genes serve as a prognostic signature and potential therapeutic target in renal cell carcinoma. Discover oncology 2026 — PMC13263375.
- Comprehensive analysis of PPP4C's impact on prognosis, immune microenvironment, and immunotherapy response in lung adenocarcinoma using single-cell sequencing and multi-omics. Frontiers in immunology 2024 — PMC11254641.
- PPP4C: a potential molecular marker and therapeutic target in thyroid cancer and triple-negative breast cancer. BMC cancer 2026 — PMC13288592.
- PubMed PMID:1336397 — UniProt-cited evidence.
- PubMed PMID:9837938 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP4C (Serine/threonine-protein phosphatase 4 catalytic subunit). Expected localisation Cytoplasmic staining, with nuclear staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06390-4). Validated IHC antibody: A06390-4 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Glandular cells); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06390-4); 2. Fixation: Image fixative and duration unreported (datasheet A06390-4); 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 A06390-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06390-4); 8. Primary antibody: Rabbit anti-PPP4C, 2-5 μg/ml (datasheet A06390-4); 9. Primary incubation: Overnight at 4 °C (datasheet A06390-4). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans residues 2–307 (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt P60510, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A06390-4, PMC11058967, PMC13263375. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP4C/immunohistochemistry-ihc-guide.