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PPP2R2A / Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform · IHC design guide
Design Immunohistochemistry for PPP2R2A
Plan PPP2R2A paraffin-section IHC around the cytoplasmic pattern seen in most tissues (HPA tissue IHC). Start the catalog antibody at 1:25 (datasheet: A04159 IHC-P), compare breast glandular cells with adipocytes (HPA tissue IHC), and account for the antibody specificity caution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP2R2A 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 | Mostly cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adipose tissue+2 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 | Antibody may detect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Broad expression; cell-type intensity varies (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; N-terminal epitope coverage is unclear (UniProt; datasheet: A04159 N-term) |
Section 1
Recommended PPP2R2A IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by four published PPP2R2A IHC protocols (PMC4409761; PMC11825944; PMC9925808; PMC7605676).
| Sample | Paraffin-embedded H. liver tissue; fixative not specified (datasheet A04159) |
| Fixation | Image fixative and duration unreported (datasheet A04159); 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-PPP2R2A, 1:25 (datasheet A04159) |
| 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 | PPP2R2A-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page antigen retrieval).
Section 2
What Is the Expected PPP2R2A Staining Pattern?
PPP2R2A is expected mainly in the cytoplasm of cells across many tissues (HPA: tissue IHC profile; UniProt P63151: expression in all tissues examined). Strong examples include breast and duodenal glandular cells, bronchial respiratory epithelial cells, caudate neurons, and pancreatic exocrine glandular cells (HPA: High). HPA rates the tissue staining Approved, with medium agreement with RNA and a warning that the antibodies may target proteins from more than one gene (HPA: reliability). The protein has no transmembrane segment (UniProt P63151: topology).
| Cytoplasmic staining in breast or duodenal glandular cells, bronchial epithelium, caudate neurons, or pancreatic exocrine cells. | This fits reported strong tissue staining (HPA: High in these cell types) and the broad cytoplasmic profile (HPA: tissue IHC). Score the relevant cells and compartment, since adjacent cell types need not share the same intensity (HPA: cell-type-specific tissue observations). |
| Predominantly nuclear or membrane-rim staining with little cytoplasmic signal. | This differs from the reported tissue pattern (HPA: cytoplasmic expression in most tissues) and the approved cytosolic ICC-IF location (HPA: subcellular). Treat it as an unconfirmed pattern: inspect morphology, compare a known-positive section, and check detection controls (general IHC practice). Absence of a transmembrane segment also gives no basis to expect membrane staining (UniProt P63151: topology). |
| Strong staining in adipocytes, liver cholangiocytes, or soft-tissue fibroblasts. | These specific cell types were reported as not detected, even though expression was seen broadly across tissues (HPA: tissue IHC; UniProt P63151: tissue specificity). Check cell identity and compare control sections (general IHC practice). Off-target binding or endogenous chromogen activity are possible explanations, not diagnoses from the image alone (general IHC practice; HPA: multi-gene antibody caution). |
| Uniform color over cells and stroma, obscuring cell boundaries. | A widespread haze does not match the reported cell-based cytoplasmic pattern (HPA: tissue IHC). Assess a no-primary control and the detection chemistry, then optimize blocking, washing, and chromogen development as needed (general IHC practice). A diffuse slide alone cannot establish PPP2R2A distribution (general IHC practice). |
| No staining in the expected cells of a breast or pancreatic positive-control section. | That conflicts with reported High staining in breast glandular cells or pancreatic exocrine cells (HPA: tissue IHC). First verify tissue morphology and that the expected cells are present; then review the antibody, detection reagents, and run controls (general IHC practice). One blank section does not establish biological absence, especially given the Approved rather than Enhanced tissue validation (HPA: antibody status and reliability). |
💡Expected PPP2R2A appearanceCall the result consistent when identifiable expected cells show predominantly cytoplasmic staining, potentially strong in HPA High examples, while isolated nuclear, membrane-rim, or stromal haze is treated as suspect until controlled (HPA: tissue IHC and subcellular; general IHC practice).
| Cell-type context | Broad expression does not imply equal staining in every cell: HPA reports High glandular, epithelial, neuronal, and exocrine examples, Low staining in some other cells, and Not detected in adipocytes, cholangiocytes, and soft-tissue fibroblasts (HPA: tissue IHC; UniProt P63151: tissue specificity). |
| Antibody attribution | Tissue IHC is Approved with medium RNA–staining consistency, but HPA cautions that the antibodies may target proteins from more than one gene (HPA: reliability). Interpret an unexpected pattern provisionally; HPA lists HPA042122, HPA042770, and CAB079289 as IHC Approved, without an Enhanced designation in the supplied record (HPA: antibodies). |
| IF/ICC: where should signal appear? | The separate ICC-IF evidence places signal in the cytosol and rates that location Approved (HPA: subcellular). Its multi-gene antibody caution also applies (HPA: subcellular). Use this as a compartment comparison for IHC, not as proof that staining in a particular paraffin-section cell is specific (HPA: tissue IHC reliability). |
| Protein features and isoforms | UniProt records 2 isoforms, no signal peptide, no propeptide, and no transmembrane segment (UniProt P63151: isoforms, processing, topology). These facts support a non-secreted, non-membrane expectation; they do not locate the antibody epitope or establish an isoform-specific staining pattern (UniProt P63151: record scope). |
| Section and detection conditions | No PPP2R2A-specific fixation sensitivity or antigen-retrieval effect is supplied (UniProt P63151 and HPA: supplied record scope). For paraffin IHC, use the validated assay instructions and matched run controls when evaluating retrieval or detection changes (general IHC practice); do not infer such effects from HPA staining levels. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank. | The run may have a detection or handling failure; a negative result alone cannot distinguish this from low target signal (general IHC practice). | Confirm that the expected cells are present and test a known-positive section in the same run; breast glandular or pancreatic exocrine cells are reported High (HPA: tissue IHC; general IHC practice). |
| The whole section has diffuse brown background. | Non-specific detection, incomplete blocking or washing, or excessive development can obscure a cell-based pattern (general IHC practice). | Examine a no-primary control, then review blocking, washes, detection, and development time before scoring (general IHC practice). |
| Nuclei dominate the staining. | A nuclear-dominant result conflicts with the reported cytoplasmic or cytosolic location; its cause is unresolved from staining alone (HPA: tissue IHC and subcellular). | Check morphology and counterstain, compare a positive section, and review detection controls; report the compartment discrepancy if it persists (general IHC practice; HPA: expected location). |
| Adipocytes or cholangiocytes stain strongly. | Those cell types were reported Not detected; cell misidentification, off-target staining, or endogenous chromogen activity could explain a discrepancy (HPA: tissue IHC and multi-gene caution; general IHC practice). | Recheck cell identity and no-primary controls, then compare staining with expected positive cells on a suitable section (general IHC practice; HPA: tissue IHC). |
| Signal appears only at cell borders. | A border-only result does not match the reported cytoplasmic pattern, and UniProt lists no transmembrane segment (HPA: tissue IHC; UniProt P63151: topology). | Review focus, tissue edges, chromogen deposition, and controls before assigning a cellular compartment (general IHC practice). |
| Two antibodies give different patterns. | The supplied IHC statuses are Approved, and HPA warns of possible multi-gene targeting; the record does not resolve which discordant pattern represents PPP2R2A (HPA: antibodies and reliability). | Compare the same expected cell types, compartment, and controls across sections; retain the disagreement in the interpretation rather than scoring either pattern as definitive (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PPP2R2A staining
A04159 · IHCImmunohistochemical analysis of paraffin-embedded H. liver section using PPP2R2A Antibody (N-term) (Cat#A04159). A04159 was diluted at 1:25
A04159 · IHCImmunohistochemical analysis of paraffin-embedded H. kidney section using PPP2R2A Antibody (N-term) (Cat#A04159). A04159 was diluted at 1:25
A04159 · IFFluorescent image of Hela cells stained with PPP2R2A Antibody (N-term). A04159 was diluted at 1:25 dilution. An Alexa Fluor 488-conjugated g
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000221914-PPP2R2A/tissue/cerebral+cortex#imid_12309077
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000221914-PPP2R2A/tissue/colon#imid_12313198
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000221914-PPP2R2A/tissue/liver#imid_12313191
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000221914-PPP2R2A/tissue/kidney#imid_12313192
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042122; green = PPP2R2A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA042122; green = PPP2R2A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP2R2A IHC & IF
🧪Run breast first; glandular cells should stain (HPA: High in breast glandular cells). Use adipose tissue as the negative tissue, where adipocytes should lack specific staining (HPA: Not detected in adipocytes); on the breast slide, cells scored as internal negatives should show counterstain without specific DAB, although the supplied HPA rows do not identify a confirmed negative breast cell type.
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP2R2A in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a control IgG matched to the primary antibody’s host species and isotype, and a PPP2R2A knockout specimen or validated peptide-block control (standard IHC practice). Check endogenous peroxidase in breast sections and endogenous biotin if using biotinylated detection (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A04159 paraffin-section caption also leaves the fixative unreported (A04159 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier, and reports no breast-specific artefact; optimize retrieval empirically and assess background in the controls (standard IHC practice).
HPA tissue IHC evidence for PPP2R2A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced PPP2R2A IHC Tips
Troubleshoot PPP2R2A staining in paraffin-section chromogenic IHC, using the selected antibody’s liver image and the stated expression evidence as context.
How should I retrieve PPP2R2A in paraffin sections when cytoplasmic staining is weak?
Use citrate buffer at pH 6.0 for heat-induced epitope retrieval at 95–98 °C for 20 minutes (page retrieval setting). If staining remains weak, compare a modestly longer heating interval on matched sections while keeping the buffer and pH fixed, then check whether tissue structure and background deteriorate (standard IHC practice). The selected antibody produced a DAB image on a paraffin-embedded human liver section at 1:25, but its caption reports neither retrieval nor fixation, so that image does not establish an optimal retrieval condition (A04159 caption). Include a positive control section in each retrieval comparison; broad tissue expression alone cannot verify antibody specificity (UniProt P63151 tissue specificity; standard IHC practice).
Could fixation explain inconsistent PPP2R2A staining between paraffin blocks?
PPP2R2A-specific fixation sensitivity is unknown from the supplied evidence: the selected paraffin-section image does not state its fixative or fixation duration (A04159 caption). Compare blocks with documented processing histories using the same citrate pH 6.0 retrieval, 95–98 °C for 20 minutes, antibody dilution, and DAB development time (page retrieval setting; standard IHC practice). If archival blocks differ, run a shared positive control beside them and inspect tissue preservation before attributing intensity differences to PPP2R2A biology (standard IHC practice). Do not infer fixation tolerance from the reported cytoplasmic tissue pattern, absence of a transmembrane segment, or annotated acetylation (HPA tissue IHC; UniProt P63151 topology and modified residues).
Should a PPP2R2A-positive IHC signal be nuclear or cytoplasmic?
Expect predominantly cytoplasmic staining in tissue sections, while treating that pattern as provisional because the tissue resource cautions that its antibody targets proteins from more than one gene (HPA tissue IHC). An independent subcellular resource reports cytosol as its approved location but carries the same cross-gene antibody caution (HPA subcellular). PPP2R2A has no annotated transmembrane segment, so a sharp membrane-only rim warrants scrutiny rather than automatic acceptance as target staining (UniProt P63151 topology; standard IHC interpretation). Compare compartment patterns in matched positive-control sections under the same citrate pH 6.0, 20-minute retrieval and DAB development conditions before interpreting a new nuclear or membrane pattern (page retrieval setting; standard IHC practice).
Can this antibody distinguish PPP2R2A isoforms or a masked N-terminal epitope?
The record lists 2 PPP2R2A isoforms, but the supplied antibody evidence does not map its binding residues or establish isoform-specific staining (UniProt P63151 isoforms; A04159 caption). The selected image identifies an N-terminal antibody and shows paraffin-section DAB staining at 1:25; that description alone cannot prove which isoform contributes to the signal (A04159 caption). PPP2R2A has an annotated N-terminal acetylation site at residue 2, yet no supplied evidence shows that this modification masks the antibody epitope (UniProt P63151 modified residues). If isoform assignment matters, obtain epitope mapping and use an orthogonal isoform-resolving assay before assigning a chromogenic signal to either isoform (standard IHC validation practice).
How can I check PPP2R2A by multiplex IF alongside chromogenic IHC?
Use the companion IF/ICC experiment to compare PPP2R2A with a marker identifying the expected cell population; pancreatic exocrine glandular cells are one reported high-staining IHC population (HPA tissue IHC). Choose fluorophores after viewing unstained tissue in each channel, placing the weaker signal in a channel with less tissue autofluorescence and including single-stain controls (standard IF practice). Because PPP2R2A lacks a transmembrane segment and cytosol is the reported location, permeabilise cells or sections sufficiently to reach the intracellular epitope, then titrate permeabilisation against morphology and background (UniProt P63151 topology; HPA subcellular; standard IF practice). Treat IF colocalisation as supporting context for the 1:25 paraffin-section DAB example, not as proof of antibody specificity (A04159 caption; standard IF interpretation).
What should I adjust when PPP2R2A DAB staining looks diffuse or nonspecific?
First inspect a no-primary control for detection-system signal and an adjacent section for pigment, folds, edge staining, or damaged tissue (standard IHC practice). For peroxidase-based DAB detection, apply an appropriate endogenous peroxidase block, then optimise protein blocking, primary-antibody concentration, washes, and DAB development on matched sections (standard IHC practice). The selected image used an undiluted biotinylated goat polyvalent secondary and 1:25 primary dilution, so a no-primary control is especially useful when evaluating that detection arrangement (A04159 caption; standard IHC practice). Keep citrate at pH 6.0 and 20 minutes constant during the first dilution comparison so retrieval changes do not confound background assessment (page retrieval setting; standard IHC practice).
How should I score PPP2R2A staining across specimens with mixed cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Score the cytoplasmic compartment within a defined cell population and record the percentage of positive cells plus intensity grades 0–3 on consistently developed DAB slides (HPA tissue IHC; standard IHC scoring practice). An H-score sums each intensity grade multiplied by its percentage of cells, giving a range of 0–300; report the scoring rule and threshold used (standard IHC scoring practice). Normalise counts or positive area to evaluable target-cell number or viable tissue area, respectively, and report cells per mm² only when density is the intended measure (standard IHC scoring practice). Keep section thickness, retrieval at citrate pH 6.0 for 20 minutes, imaging, and DAB development comparable across specimens (page retrieval setting; standard IHC practice).
How do I distinguish convincing PPP2R2A staining from a chromogenic artefact?
A plausible result shows cytoplasmic staining in intact cells, consistent with the reported tissue pattern, while the antibody cross-gene caution limits target-specific conclusions (HPA tissue IHC). Compare a reported high-staining population, such as breast glandular cells, with a reported undetected population, such as adipocytes, without treating either as an absolute biological control (HPA tissue IHC). Reject edge-restricted staining, necrotic regions, and signal reproduced in a no-primary or endogenous-peroxidase control as evidence of PPP2R2A expression (standard IHC practice). The selected 1:25 paraffin-section DAB image demonstrates a staining example, but orthogonal validation is needed before assigning unexpected nuclear, membrane-only, or cell-specific signal to PPP2R2A (A04159 caption; HPA subcellular; standard IHC interpretation).
Boster reagents
Best PPP2R2A / Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform IHC Antibodies
The catalog antibody has IHC-P images of human liver and kidney paraffin sections and an IF image of HeLa cells (A04159 image captions; catalog: Human reactivity).
Real IHC data
Anti-PPP2R2A Antibody (N-term)
Cat # A04159
A04159 is listed for IHC-P and IF in human samples (catalog: applications and reactivity). Its images show paraffin sections of human liver and kidney stained by IHC, plus HeLa cells stained by IF (A04159 image captions).
Which to pick: Choose A04159 for tissue IHC: it is a rabbit polyclonal antibody listed for IHC-P, with paraffin-section images at 1:25; the fixative is unreported (A04159 catalog; IHC image captions). For IF in cells, A04159 is listed for IF and has a HeLa-cell image at 1:25; ICC validation is not separately reported (A04159 catalog; IF image caption). No cross-species option is documented because A04159 lists Human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P63151 (2ABA_HUMAN, Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform).
- Human Protein Atlas. PPP2R2A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PPP2R2A subcellular location (ICC-IF): Localized to the cytosol. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PPP2R2A antibody validation summary (3 antibodies).
- Altered PPP2R2A and Cyclin D1 expression defines a subgroup of aggressive luminal-like breast cancer. BMC cancer 2015 — PMC4409761.
- Characterization of RNF144B and PPP2R2A identified by a novel approach using TCGA data in ovarian cancer. Scientific reports 2025 — PMC11825944.
- VPA mediates bidirectional regulation of cell cycle progression through the PPP2R2A-Chk1 signaling axis in response to HU. Cell death & disease 2023 — PMC9925808.
- Hsa-MiR-590-3p Promotes the Malignancy Progression of Pancreatic Ductal Carcinoma by Inhibiting the Expression of p27 and PPP2R2A via G1/S Cell Cycle Pathway. OncoTargets and therapy 2020 — PMC7605676.
- PubMed PMID:1849734 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16421571 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP2R2A (Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B alpha isoform). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04159 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded H. liver tissue; fixative not specified (datasheet A04159); 2. Fixation: Image fixative and duration unreported (datasheet A04159); 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-PPP2R2A, 1:25 (datasheet A04159); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody may detect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; N-terminal epitope coverage is unclear (UniProt; datasheet: A04159 N-term); Regulation: Broad expression; cell-type intensity varies (HPA tissue IHC). Evidence: UniProt P63151, HPA tissue IHC · Approved, HPA ICC-IF · cytosol. Caution: Based on antibodies targeting proteins from…, Datasheet A04159, PMC4409761, PMC11825944. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP2R2A/immunohistochemistry-ihc-guide.