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PPP2CB / Serine/threonine-protein phosphatase 2A catalytic subunit beta isoform · IHC design guide
Design Immunohistochemistry for PPP2CB
Plan PPP2CB chromogenic IHC in paraffin sections using the reported cytoplasmic and occasional nuclear staining pattern (HPA tissue IHC). Start antibody A07661 at 1:100–1:200 (datasheet: A07661 IHC), and interpret staining with the reported cross-gene caution in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP2CB 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; nuclear in some tissues (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining; nuclear in some tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific) | |
| Caveat | Staining may include protein from another gene (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | No isoforms or cleavage; intracellular epitope access matters (UniProt) |
Section 1
Recommended PPP2CB IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published PPP2CB protocol using paraffin-embedded colon cancer sections (PMC8377503 methods).
| Sample | Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A07661) |
| Fixation | Image fixative and duration unreported (datasheet A07661); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-PPP2CB, 1:100-1:200 (datasheet A07661) |
| 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 | PPP2CB-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression and nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule; UniProt P62714: nucleus). The article does not specify a heat-retrieval method (PMC8377503 methods).
Section 2
What Is the Expected PPP2CB Staining Pattern?
PPP2CB is a soluble protein found in the cytoplasm and nucleus, with centromere and spindle-pole localization during mitosis (UniProt P62714 topology and subcellular location). In paraffin-section IHC, expect broad cytoplasmic staining and nuclear staining in some tissues, including strong staining in several epithelial, hematopoietic, and neuronal cell populations (HPA tissue IHC). Interpret intensity cautiously: HPA rates the tissue pattern Approved, with medium agreement with RNA data and a warning that staining may reflect more than one gene (HPA tissue IHC).
| Clear cytoplasmic staining, with nuclei stained in some cells. | This fits the broad tissue pattern reported by HPA and the cytoplasmic and nuclear locations annotated by UniProt (HPA tissue IHC; UniProt P62714 subcellular location). Judge the signal against cell morphology and adjacent background; nuclear staining need not be equally prominent in every tissue (HPA tissue IHC). |
| Strong staining in adrenal gland or appendix glandular cells, bone marrow hematopoietic cells, or cortical neurons. | These are documented high-staining cell populations and useful positive tissue references (HPA tissue IHC: High in each listed population). A strong result supports assay performance, but staining alone cannot establish that the antibody distinguishes PPP2CB from other gene products (HPA tissue IHC: multi-gene caution). |
| A sharp cell-surface rim or predominantly extracellular deposit replaces the expected intracellular signal. | That distribution conflicts with the annotated cytoplasmic and nuclear locations and the absence of a transmembrane segment (UniProt P62714 subcellular location and topology). Treat it as suspect; check section morphology, precipitate, and control slides before assigning it to PPP2CB. |
| Strong signal appears chiefly in cardiomyocytes, skeletal myocytes, or soft-tissue fibroblasts. | HPA reports Low staining in those populations, not absence (HPA tissue IHC). Compare them with a documented high-staining population in the same run. Unexpected relative intensity warrants checks for nonspecific binding or endogenous detection activity; it does not by itself prove cross-reactivity. |
| Diffuse chromogen covers cells and extracellular space, or a documented high-staining tissue has no signal. | A uniform haze obscures compartment scoring; absence in a high-staining reference weakens confidence in a negative test section (HPA tissue IHC: High reference populations). Use control slides to distinguish background or detection failure from a plausible biological difference; neither appearance alone identifies the cause. |
💡Expected PPP2CB appearanceCall a section positive when identifiable cells show cytoplasmic staining, with nuclear staining where present, and a documented high-staining population is clearly above local background (HPA tissue IHC; UniProt P62714 subcellular location); isolated membrane rims, extracellular deposits, or uniform haze are suspect.
| Cell population and reference tissue | HPA reports High staining in adrenal and appendix glandular cells, bone marrow hematopoietic cells, and several neuronal populations, but Low staining in cardiomyocytes, skeletal myocytes, and soft-tissue fibroblasts (HPA tissue IHC). Use these as relative intensity references; Low is not a negative control. |
| Compartment and cell-cycle state | Cytoplasm and nucleus are annotated locations; centromere localization is reported in prometaphase but not anaphase, and spindle-pole localization occurs during mitosis (UniProt P62714 subcellular location). Those transient structures should not be required in routine sections, where mitotic cells may be scarce. |
| Antibody-level interpretation | Two listed antibodies have Approved IHC status, while the tissue summary has medium agreement with RNA data and cautions that staining may reflect more than one gene (HPA antibodies; HPA tissue IHC). A plausible pattern supports interpretation but does not establish PPP2CB-specific recognition. |
| Target structure and processing | PPP2CB has no transmembrane segment, signal peptide, or propeptide; its annotated chain spans residues 1–309 (UniProt P62714 topology and processing). These facts support intracellular interpretation, but they do not establish an antibody epitope or predict antigen-retrieval conditions. |
| Fixation and antigen retrieval | Target-specific fixation sensitivity and retrieval requirements are unreported in the supplied UniProt and HPA records. For paraffin-section IHC, evaluate retrieval using the antibody’s IHC protocol and a documented high-staining tissue; do not infer a PPP2CB-specific fixation effect from tissue intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal appears in the test section or a documented high-staining reference. | The run may have failed at retrieval, primary-antibody binding, or chromogenic detection; the slide alone cannot distinguish these causes. | Check the positive control and reagent sequence, then review the catalog antibody’s IHC-P conditions before adjusting retrieval or primary dilution (HPA tissue IHC: High reference populations). |
| The whole section has a uniform haze or extracellular chromogen. | Background from incomplete blocking, washing, or detection chemistry can obscure intracellular staining (standard IHC practice). | Review the no-primary control, wash and blocking steps, and chromogen development; score only signal resolved within identifiable cells (standard IHC practice). |
| Strong staining is concentrated in a population HPA calls Low. | Relative intensity differs from the HPA reference; nonspecific binding or endogenous detection activity is possible, but Low does not mean absent (HPA tissue IHC). | Compare with a documented High population, inspect the no-primary control, and assess whether signal tracks the expected cytoplasmic or nuclear compartments (HPA tissue IHC; UniProt P62714 subcellular location). |
| Signal forms a membrane rim with little intracellular staining. | A dominant membrane pattern is difficult to reconcile with PPP2CB’s annotated locations and lack of a transmembrane segment (UniProt P62714 topology and subcellular location). | Inspect section edges and precipitate, compare control slides, and withhold a PPP2CB-positive call until an intracellular pattern is demonstrable. |
| Nuclear staining is absent, but cytoplasmic staining is clear. | HPA describes nuclear expression in several tissues rather than all tissues; UniProt also lists cytoplasmic localization (HPA tissue IHC; UniProt P62714 subcellular location). | Assess the cytoplasmic pattern and positive reference first. Record the nuclear result separately, without treating its absence alone as assay failure. |
| Does IF/ICC require the same visible balance of cytoplasmic and nuclear signal as IHC? | The HPA ICC-IF summary reports supported nucleoplasmic localization, while the tissue IHC summary reports broad cytoplasmic and some nuclear expression (HPA subcellular; HPA tissue IHC). | Interpret IF/ICC against its supported nucleoplasmic reference and IHC against the tissue pattern; use the separate IF/ICC guide for assay design (HPA subcellular; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing PPP2CB staining
A07661 · IHCImmunohistochemistry of paraffin-embedded rat kidney using PP2A Catalytic β antibody at dilution of 1:100 .Perform microwave antigen retriev
A07661 · IHCImmunohistochemistry of paraffin-embedded human colon carcinoma using PP2A Catalytic β antibody at dilution of 1:100 .Perform microwave anti
M07661 · IHCImmunohistochemical analysis of paraffin-embedded human colon, using PP2A alpha + beta Antibody.
A07661-1 · IFICC staining PP2A alpha + beta in PANC-1 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, perm
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104695-PPP2CB/tissue/cerebral+cortex#imid_12596377
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000104695-PPP2CB/tissue/esophagus#imid_12596341
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104695-PPP2CB/tissue/colon#imid_12596295
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104695-PPP2CB/tissue/liver#imid_12596296
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB018600; green = PPP2CB, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB018600; green = PPP2CB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP2CB IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain strongly (HPA: High in bone marrow hematopoietic cells). HPA detects PPP2CB in all 45 scored tissues, so there is no supported negative tissue; no-primary and isotype sections provide the negative controls, and cells on the positive slide should show no specific staining in those control sections (HPA: no negative tissue rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: PPP2CB is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP2CB in A-431, U-251MG, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an isotype control matched to the primary antibody’s host species, immunoglobulin class and concentration; use PPP2CB knockout material as a biological negative where available (standard IHC practice). Block endogenous peroxidase in bone marrow before chromogenic detection because hematopoietic cells can produce peroxidase-related background (HPA: High in bone marrow hematopoietic cells; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07661 rat-kidney paraffin-section caption does not state a fixative (A07661 tissue-IHC caption). That caption uses microwave retrieval in 10 mM PBS, pH 7.2, but does not establish that retrieval is required for bone marrow (A07661 tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF would be easier; for bone marrow chromogenic IHC, endogenous peroxidase is a practical background risk (HPA: High in bone marrow hematopoietic cells; standard IHC practice).
HPA tissue IHC evidence for PPP2CB
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PPP2CB is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PPP2CB IHC Tips
Use compartment, cell type, and matched controls to troubleshoot PPP2CB staining in paraffin sections; interpret tissue patterns cautiously because the reference antibody may recognize more than one gene (HPA).
How should I adjust retrieval when PPP2CB staining is weak in paraffin sections?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). If staining remains weak, test a separate section with 10 mM PBS, pH 7.2, using microwave retrieval; that condition accompanied 1:100 staining of paraffin-embedded rat kidney with A07661 (A07661 tissue-IHC caption). Keep section thickness, primary incubation, and chromogen development matched so retrieval is the main changed variable (standard IHC practice). Compare signal in intact cytoplasm and nuclei, both reported locations, while checking whether stronger retrieval also increases nonspecific staining or damages morphology (UniProt P62714 localisation; HPA tissue-IHC profile; standard IHC practice).
Could fixation explain uneven PPP2CB staining across my paraffin sections?
PPP2CB-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (A07661 tissue-IHC caption). Record the fixative, fixation duration, specimen thickness, and processing history for each section before comparing staining (standard IHC practice). If those histories differ, stain adjacent sections in one run with the same Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 minutes (page retrieval setting; standard IHC practice). Compare preserved morphology and within-section staining patterns, then test a different retrieval condition on matched material if needed; neither tissue distribution nor PPP2CB modifications establish a fixation effect (HPA tissue-IHC profile; UniProt P62714 modified residues; standard IHC practice).
Is predominantly nuclear PPP2CB staining credible when I expected cytoplasmic signal?
Yes: PPP2CB is annotated in both cytoplasm and nucleus, while tissue IHC reports ubiquitous cytoplasmic staining and nuclear staining in several tissues (UniProt P62714 localisation; HPA tissue-IHC profile). Nucleoplasmic localisation is also supported by subcellular imaging, so nuclear signal alone is not grounds for rejection (HPA subcellular). Score nuclear and cytoplasmic staining separately within the same identified cell population, using matched exposure and chromogen development across sections (standard IHC practice). Centromere localisation is described in prometaphase but not anaphase, and spindle-pole localisation occurs during mitosis; evaluate such puncta only in morphologically appropriate dividing cells (UniProt P62714 localisation; standard IHC practice).
Can this stain distinguish PPP2CB from related PP2A catalytic proteins?
Do not infer gene-level specificity from a positive chromogenic stain: the tissue reference warns that its antibody targets proteins from more than one gene (HPA tissue-IHC reliability description). The supplied PPP2CB record lists 0 alternative isoforms, but that annotation does not establish the catalog antibody's epitope or exclude recognition of related proteins (UniProt P62714 isoforms; standard antibody validation practice). PPP2CB has a phosphotyrosine at residue 307 and a methylated leucine at 309; modification-dependent recognition is plausible only if the mapped epitope includes those residues (UniProt P62714 modified residues; standard immunochemistry practice). Check the antibody's disclosed immunogen and independent specificity controls before assigning compartment differences to PPP2CB itself (standard antibody validation practice).
How can IF help resolve ambiguous PPP2CB cell and compartment staining?
Use IF as a separate validation experiment and multiplex PPP2CB with a marker independently validated for the cell population being scored (standard IF practice). This matters when comparing a proposed endothelial signal with the high staining reported for colon endothelial cells, for example (HPA: High in colon endothelial cells). Choose a spectrally separated far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and include single-label and unstained controls (standard IF practice). PPP2CB lacks a transmembrane segment and is reported in cytoplasm and nucleus; use controlled intracellular permeabilisation, such as 0.1% Triton X-100 for 5 minutes, then assess morphology and background (UniProt P62714 topology and localisation; standard IF practice).
What should I check when PPP2CB chromogenic staining is widespread or granular?
Begin with a no-primary control and inspect staining in tissue edges, damaged regions, and extracellular spaces before adjusting antibody concentration (standard IHC practice). Block endogenous peroxidase, for example with 3% hydrogen peroxide for 10 minutes, when using peroxidase detection; this is a general chromogenic workflow step, not PPP2CB-specific evidence (standard IHC practice). Titrate around the reported 1:100 dilution for A07661 while keeping retrieval and detection constant, and stop DAB development at a matched endpoint (A07661 tissue-IHC caption; standard IHC practice). Because PPP2CB can be cytoplasmic and nuclear, broad cellular signal is possible, but control-positive haze should not be scored as antigen staining (UniProt P62714 localisation; standard IHC practice).
How should I quantify PPP2CB staining across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since tissue IHC reports cytoplasmic expression broadly and nuclear expression in several tissues (HPA tissue-IHC profile). For a chosen population, record the percentage of positive cells and staining intensity, or calculate a compartment-specific H-score from 0 to 300 using percentages at intensity levels 0–3 (standard IHC scoring practice). For sparse populations, report positive cells per mm² of evaluable tissue and the denominator used (standard IHC scoring practice). Normalise comparisons to the same cell type, compartment, tissue area, staining batch, and scoring threshold; exclude folds, necrosis, and edges consistently (standard IHC scoring practice).
Which staining patterns would make a PPP2CB positive call unreliable?
A credible call should fit intact-cell morphology and a plausible cytoplasmic or nuclear compartment; PPP2CB has both reported locations (UniProt P62714 localisation; HPA tissue-IHC profile). Treat signal confined to cut edges, necrosis, extracellular deposits, or a no-primary control as suspect, and check whether endogenous peroxidase persists after blocking (standard IHC practice). An apparent strong signal restricted to cardiomyocytes or skeletal myocytes warrants scrutiny because those cells are listed with low staining in the tissue reference, although low is not absent (HPA: Low in cardiomyocytes and skeletal myocytes). Even a matching pattern cannot prove PPP2CB specificity because the reference tissue antibody may recognize proteins from more than one gene (HPA tissue-IHC reliability description).
Boster reagents
Best PPP2CB / Serine/threonine-protein phosphatase 2A catalytic subunit beta isoform IHC Antibodies
The catalog includes antibodies with IHC images from paraffin-embedded rat kidney, mouse kidney, human colon carcinoma, and human colon, plus an ICC image from PANC-1 cells (catalog image captions).
Real IHC data
Anti-PPP2CB Antibody
Cat # A07661
Real IHC data
Anti-PP2A alpha + beta Rabbit Monoclonal Antibody
Cat # M07661
Real IF data
Anti-PP2A Alpha / Beta PPP2CB Antibody
Cat # A07661-1
A07661 has IHC images from paraffin-embedded rat kidney, mouse kidney, and human colon carcinoma (A07661 image captions); M07661 has an IHC image from paraffin-embedded human colon (M07661 image caption). A07661-1 lists IHC and ICC applications and has an ICC image from PANC-1 cells fixed in paraformaldehyde (A07661-1 catalog; A07661-1 image caption).
Which to pick: For tissue IHC, choose A07661 when its rat kidney, mouse kidney, or human colon carcinoma examples match your sample (A07661 image captions), or M07661 for a rabbit monoclonal with a human colon example (M07661 catalog; M07661 image caption); both IHC captions report paraffin embedding but do not report the fixative (A07661 and M07661 image captions). For ICC, A07661-1 has an image from paraformaldehyde-fixed, permeabilised PANC-1 cells (A07661-1 image caption); for IF, A07661 and M07661 list that application, though their supplied figures show IHC (A07661 and M07661 catalogs; image captions). For broader listed species coverage, A07661-1 includes zebrafish alongside human, mouse, and rat, while its supplied figure documents ICC in PANC-1 cells (A07661-1 catalog; A07661-1 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62714 (PP2AB_HUMAN, Serine/threonine-protein phosphatase 2A catalytic subunit beta isoform).
- Human Protein Atlas. PPP2CB tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PPP2CB subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. PPP2CB antibody validation summary (2 antibodies).
- Comprehensive Analysis of Glycolysis-Related Genes for Prognosis, Immune Features, and Candidate Drug Development in Colon Cancer. Frontiers in cell and developmental biology 2021 — PMC8377503.
- PubMed PMID:2849765 — UniProt-cited evidence.
- PubMed PMID:2837763 — UniProt-cited evidence.
- PubMed PMID:1846293 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP2CB (Serine/threonine-protein phosphatase 2A catalytic subunit beta isoform). Expected localisation Cytoplasmic; nuclear in some tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07661 at 1:100-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat kidney tissue; fixative not specified (datasheet A07661); 2. Fixation: Image fixative and duration unreported (datasheet A07661); 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 Tris-EDTA buffer, pH 9.0 , 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-PPP2CB, 1:100-1:200 (datasheet A07661); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent. (standard IHC practice; not target-specific); Caveat: Staining may include protein from another gene (HPA tissue IHC); Isoform / epitope: No isoforms or cleavage; intracellular epitope access matters (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt P62714, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A07661, PMC8377503. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP2CB/immunohistochemistry-ihc-guide.