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PPP3CB / Serine/threonine-protein phosphatase 2B catalytic subunit beta isoform · IHC design guide
Design Immunohistochemistry for PPP3CB
Plan PPP3CB chromogenic IHC in paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A07303-1). Compare cytoplasmic staining with HPA tissue observations, keeping its uncertain reliability in mind (UniProt; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP3CB 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 (UniProt); extracellular signal in some tissues (HPA tissue IHC) | |
| Staining pattern | Macrophages and neurons: cytoplasmic; some extracellular signal (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A07303-1) | |
| Positive control | Breast+4 more · see all | |
| Negative control | Adrenal gland+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07303-1) | |
| Caveat | Tissue staining has low concordance with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | Four isoforms; check epitope coverage (UniProt) |
Section 1
Recommended PPP3CB IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A07303-1). The published options cover glioma and pancreatic tissue microarrays (PMC10733678; PMC7785039).
| Sample | Paraffin-embedded human rectum adenocarcinoma tissue; fixative not specified (datasheet A07303-1) |
| Fixation | Image fixative and duration unreported (datasheet A07303-1); 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 A07303-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A07303-1) |
| Primary antibody | Rabbit anti-PPP3CB, 2-5 μg/ml (datasheet A07303-1) |
| Primary incubation | Overnight at 4 °C (datasheet A07303-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A07303-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PPP3CB-positive staining in glandular cells of breast (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with extracellular positivity in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A07303-1); citrate pH 6.0 is a published alternative (PMC7785039).
Section 2
What Is the Expected PPP3CB Staining Pattern?
PPP3CB is expected mainly in the cytoplasm, consistent with its lack of a transmembrane segment (UniProt P16298: cytoplasm, topology). HPA reports high IHC staining in breast glandular cells, neurons in several brain regions, and lung macrophages (HPA: tissue IHC). Use these as candidate positive contexts, with caution: HPA rates the tissue staining pattern Uncertain because antibody staining and RNA expression have low consistency (HPA: reliability).
| Cytoplasmic staining in breast glandular cells, neurons, or lung macrophages, with distinguishable cellular boundaries. | This fits the expected compartment (UniProt P16298: cytoplasm) and cells reported as High by HPA (HPA: tissue IHC). Score intensity in the specified cells, not the whole section; the match is supportive, while HPA's Uncertain tissue reliability limits confirmation of specificity (HPA: reliability). |
| Signal is predominantly confined to nuclei or cell membranes, with little cytoplasmic staining. | This conflicts with the principal cytoplasmic assignment and lacks support from UniProt topology (UniProt P16298: cytoplasm, no transmembrane segment). Review morphology and controls before calling it PPP3CB. HPA's ICC-IF record lists nucleoplasm as an uncertain additional location, so occasional nuclear signal alone is inconclusive (HPA: subcellular). |
| Strong staining appears in an unexpected cell population within a section. | First identify the stained cells by morphology and compare them with the relevant HPA cell-specific entry (HPA: tissue IHC). A mismatch may reflect nonspecific binding or endogenous chromogenic detection activity (general IHC practice). It cannot establish a new PPP3CB cell distribution without independent validation, especially given HPA's Uncertain rating (HPA: reliability). |
| Broad haze obscures cells, or extracellular material stains more strongly than cell cytoplasm. | Treat diffuse haze as background until controls and tissue structure clarify it (general IHC practice). Extracellular positivity occurs in several HPA tissues, so extracellular signal is not automatically an artefact; it is also insufficient on its own to establish a cellular PPP3CB pattern (HPA: tissue IHC profile). |
| No visible staining occurs in a candidate positive tissue. | A blank result in a region reported as High by HPA calls for a technical check, including section integrity, antibody detection, and the assay's retrieval conditions (HPA: tissue IHC; general IHC practice). It does not prove absence of PPP3CB, and the HPA reference pattern itself remains Uncertain (HPA: reliability). |
💡Expected PPP3CB appearanceCall a result consistent when cytoplasmic staining is clear in an HPA High cell population, such as lung macrophages, breast glandular cells, or neurons; predominantly membrane-only staining or diffuse haze is suspect, subject to controls (UniProt P16298: cytoplasm, topology; HPA: tissue IHC; general IHC practice).
| Compartment and topology | UniProt places PPP3CB in the cytoplasm and reports no transmembrane segment (UniProt P16298). Use cellular cytoplasm as the main localization criterion; the record gives no basis to expect a membrane-rim pattern. |
| Cell-specific reference patterns | HPA reports High staining in breast glandular cells, caudate, cortical and hippocampal neurons, and lung macrophages; it reports Medium staining in cerebellar Purkinje cells (HPA: tissue IHC). Compare like cell types when judging intensity, rather than treating all cells in a tissue as equivalent. |
| Strength of the reference evidence | HPA marks tissue IHC reliability Uncertain because staining and RNA expression show low consistency, with external verification pending; HPA008233 is also IHC Uncertain (HPA: reliability, antibodies). A pattern match supports interpretation but does not independently validate antibody specificity. |
| Isoforms and processing | UniProt lists four PPP3CB isoforms and a chain spanning residues 2–524, with no signal peptide or propeptide (UniProt P16298). The supplied record gives no antibody epitope, so it cannot establish which isoforms an IHC antibody detects or whether isoform differences alter staining. |
| IF/ICC Q&A: should the same localization be expected? | Mainly cytosolic staining is supported in HPA ICC-IF; nucleoplasm is an uncertain additional location and calyx is approved (HPA: subcellular). HPA cautions that this annotation uses antibodies targeting proteins from multiple genes, so it should not override the uncertain tissue IHC assessment (HPA: subcellular, reliability). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in an HPA High cell population. | The result may reflect an assay failure or absent detectable signal; HPA's Uncertain reference cannot resolve which (HPA: tissue IHC, reliability). | Check section morphology and a working detection control, then review the antibody's IHC dilution and retrieval conditions against its supplied instructions (general IHC practice). Do not infer a PPP3CB-specific fixation effect from this result. |
| All cell types or the entire section show a uniform chromogenic tint. | Widespread tint may arise from background binding or endogenous detection activity (general IHC practice). | Compare a no-primary control; review blocking, washes, and detection chemistry, then reassess whether cytoplasmic signal remains in the expected cells (general IHC practice; UniProt P16298: cytoplasm; HPA: tissue IHC). |
| Predominantly nuclear or membrane-rim staining persists. | That pattern conflicts with the main cytoplasmic assignment; HPA lists nucleoplasm only as an uncertain additional ICC-IF location (UniProt P16298: cytoplasm, topology; HPA: subcellular). | Recheck cell boundaries and counterstain, compare controls, and seek independent specificity evidence before scoring the pattern as PPP3CB (general IHC practice; HPA: reliability Uncertain). |
| Extracellular staining dominates the section. | HPA reports extracellular positivity in several tissues, but does not establish that this alone identifies the source of PPP3CB signal (HPA: tissue IHC profile). | Score intracellular staining separately from extracellular deposits; inspect a no-primary control and report the extracellular observation without treating it as cell-specific evidence (general IHC practice; HPA: tissue IHC profile). |
| A nominally negative comparator stains strongly. | HPA lists adrenal gland glandular cells, ovarian follicle cells, and parathyroid gland glandular cells as Not detected, yet rates the overall tissue pattern Uncertain (HPA: tissue IHC, reliability). | Confirm the actual stained cell type, review the no-primary control, and seek orthogonal validation before interpreting unexpected staining as PPP3CB expression (general IHC practice; HPA: reliability). |
| The expected cytoplasmic cells stain, but the signal is hard to separate from background. | Weak contrast may limit cell-level scoring, while HPA's Uncertain rating limits how much the reference pattern can settle specificity (general IHC practice; HPA: reliability). | Compare matched controls and cell morphology; optimize general IHC conditions within the antibody's validated instructions, then document the cell-specific result and any remaining uncertainty (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing PPP3CB staining
A07303-1 · IHCIHC analysis of PPP3CB using anti-PPP3CB antibody (A07303-1). PPP3CB was detected in a paraffin-embedded section of human rectum adenocarcin
A07303-1 · IHCIHC analysis of PPP3CB using anti-PPP3CB antibody (A07303-1). PPP3CB was detected in a paraffin-embedded section of human tonsil tissue. Hea
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107758-PPP3CB/tissue/cerebral+cortex#imid_2668632
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107758-PPP3CB/tissue/colon#imid_2668566
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107758-PPP3CB/tissue/liver#imid_2668567
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107758-PPP3CB/tissue/kidney#imid_2668560
HPA · ICC-IF GAMGHuman Protein Atlas immunofluorescence, GAMG, antibody HPA074370; green = PPP3CB, blue = nucleus, red = microtubules.
HPA · ICC-IF GAMGHuman Protein Atlas immunofluorescence, GAMG, antibody HPA074370; green = PPP3CB, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP3CB IHC & IF
🧪Run lung first: macrophages should stain strongly (HPA: High in lung macrophages). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected); on the lung slide, adjacent nonmacrophage cells should be assessed for background rather than assumed to be PPP3CB-negative (HPA: High in lung macrophages).
Positive control tissue: Breast (Glandular cells, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP3CB in GAMG, PC-3, U2OS, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG control of the same antibody format (selected-SKU caption: rabbit primary antibody); confirm specificity with PPP3CB knockout material or a peptide block if the immunizing peptide is available. Quench endogenous peroxidase and distinguish DAB signal from pigment in lung macrophages (HPA: High in lung macrophages; selected-SKU caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A07303-1 paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but retrieval dependence and whether frozen sections or IF are easier are unreported (selected-SKU caption: EDTA retrieval). For IF/ICC, cytosol is the supported location, though HPA cautions that its localization evidence uses antibodies targeting proteins from multiple genes; lung macrophage autofluorescence may complicate interpretation (HPA subcellular: cytosol supported; HPA: High in lung macrophages).
HPA tissue IHC evidence for PPP3CB
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
Section 3
Advanced PPP3CB IHC Tips
Troubleshoot PPP3CB staining in paraffin sections by checking retrieval, cell type, compartment and controls before comparing chromogenic IHC signals (UniProt P16298; HPA tissue IHC).
Which retrieval conditions should I try first for weak PPP3CB staining?
Start with heat-mediated retrieval in EDTA buffer at pH 8.0 (datasheet A07303-1). The selected paraffin-section example used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (datasheet A07303-1). If staining is weak, compare a shorter and longer heating exposure on adjacent sections while holding antibody concentration, detection and development constant (standard IHC practice). Include a tissue compartment with reported PPP3CB positivity, such as lung macrophages, to judge whether the change improves specific staining (HPA: High in lung macrophages). Record tissue damage and background alongside signal, since stronger retrieval alone does not establish specificity (standard IHC practice).
How should I troubleshoot fixation-related loss of PPP3CB signal?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet A07303-1). Document the actual fixative, fixation duration, processing history and section age for each specimen before comparing staining (standard IHC practice). Run differently processed sections through the same EDTA pH 8.0 retrieval and 2 μg/ml primary-antibody conditions used in the example, if those conditions suit the comparison (datasheet A07303-1). Compare preserved morphology, internal positive cells and negative controls across sections, and change one processing variable at a time (standard IHC practice). Neither tissue staining patterns nor PPP3CB sequence features establish a fixation-specific effect (HPA tissue IHC; UniProt P16298).
Where should convincing PPP3CB chromogenic staining appear?
Prioritise cellular cytoplasmic staining when evaluating the DAB signal, because PPP3CB is annotated in the cytoplasm and lacks a transmembrane segment (UniProt P16298 topology). The subcellular atlas supports cytosol as the main location, while its nucleoplasmic assignment is uncertain and its antibodies may target proteins from multiple genes (HPA subcellular). Inspect individual cells against the counterstain so diffuse extracellular colour is not scored as intracellular signal (standard IHC practice). Compare the compartment pattern with positive cell populations on the same run; lung macrophages and cortical neuronal cells have reported High staining (HPA tissue IHC). Unexpected nuclear-only or membrane-rim staining warrants control review before biological interpretation (UniProt P16298; standard IHC practice).
Can this staining distinguish PPP3CB isoforms or a modified epitope?
Do not assign an IHC signal to a single splice form without epitope mapping: PPP3CB has 4 annotated isoforms, and the supplied caption does not identify the antibody epitope (UniProt P16298; datasheet A07303-1). The record lists N-acetylalanine at residue 2 and phosphoserine at residue 478, but provides no evidence that either modification changes this antibody’s staining (UniProt P16298). Request or inspect the immunogen sequence and compare it with the isoform sequences before making isoform claims (standard antibody-validation practice). Use an independent antibody against a distinct known epitope, where available, to investigate discordant patterns (standard IHC practice). Report the observed staining as PPP3CB immunoreactivity until specificity is established (standard IHC practice).
How should I adapt the localisation check for a separate IF experiment?
Treat IF/ICC as a separate validation experiment; the selected product example documents chromogenic staining in a paraffin section, with no IF conditions supplied (datasheet A07303-1). Multiplex PPP3CB with an independently validated marker for the expected cell population, such as a macrophage marker when examining lung macrophages (HPA: High in lung macrophages; standard IF practice). Select a fluorophore channel after checking tissue autofluorescence with an unstained control, and include single-stain controls to assess bleed-through (standard IF practice). For an intracellular epitope, assess a permeabilisation condition that permits access to cytosolic PPP3CB, which has no transmembrane segment (UniProt P16298 topology; standard IF practice). Keep IF fixation and permeabilisation conclusions separate from the paraffin-section IHC caption, whose fixative is unreported (datasheet A07303-1).
How can I separate PPP3CB signal from chromogenic background?
Inspect a no-primary control for secondary-reagent binding and endogenous peroxidase colour before interpreting brown deposits (standard IHC practice). Include a peroxidase-blocking step in the HRP workflow, and compare DAB development times across matched sections (standard IHC practice). The selected example used 10% goat-serum block, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A07303-1). If diffuse colour persists, optimise block, primary concentration and wash stringency one variable at a time (standard IHC practice). Do not use extracellular positivity alone to establish specificity, given the reported extracellular pattern and uncertain tissue-IHC reliability (HPA tissue IHC).
What should I score when comparing PPP3CB across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic scoring compartment before reviewing experimental groups (UniProt P16298 cytoplasm; standard IHC practice). For a cell-based comparison, report the percentage of positive cells and an H-score combining staining intensity with the percentage at each intensity; keep the scoring thresholds fixed across slides (standard IHC practice). Where the question concerns infiltrating positive cells, report their density per mm² of viable tissue as well (standard IHC practice). Normalise to the same eligible cell population or viable tissue area, and record section quality and staining batch (standard IHC practice). Interpret differences cautiously because atlas tissue-IHC reliability is Uncertain owing to low staining–RNA consistency (HPA tissue IHC).
Which apparent PPP3CB positives warrant a closer artefact check?
A convincing result should show intracellular staining in identifiable cells with the expected cytoplasmic emphasis and appropriate control behaviour (UniProt P16298 cytoplasm; standard IHC practice). Review nuclear-only deposits cautiously because nucleoplasmic localisation is uncertain in the subcellular atlas, which also cautions that its antibodies may target proteins from multiple genes (HPA subcellular). Check whether the claimed positive cell type fits the reference pattern; lung macrophages are reported High, while adrenal glandular cells are Not detected (HPA tissue IHC). Exclude edge staining, necrotic areas and colour reproduced in a no-primary or peroxidase-control section from the scored signal (standard IHC practice). Treat unexpected extracellular staining cautiously because the atlas reports it alongside Uncertain tissue-IHC reliability (HPA tissue IHC).
Boster reagents
Best PPP3CB / Serine/threonine-protein phosphatase 2B catalytic subunit beta isoform IHC Antibodies
The IHC-validated anti-PPP3CB antibody has paraffin-section images from human rectum adenocarcinoma, tonsil and urothelial carcinoma (catalog IHC captions); its listed reactivity also includes monkey, mouse and rat (catalog reactivity).
Real IHC data
Anti-PPP3CB Antibody ®
Cat # A07303-1
A07303-1 is the sole SKU shown, with IHC images from paraffin sections of human rectum adenocarcinoma, tonsil and urothelial carcinoma (catalog IHC captions). Its application list includes IHC but not IF/ICC, and its listed reactivity is human, monkey, mouse and rat (catalog applications and reactivity).
Which to pick: Choose A07303-1 for human tissue IHC on paraffin sections; its captions document 2 μg/ml antibody after heat retrieval in EDTA, pH 8.0, but do not report the fixative (A07303-1 IHC captions). There is no IF/ICC-validated SKU in this payload (catalog applications; catalog IF images: none). For work in monkey, mouse or rat, A07303-1 lists reactivity with those species, while the supplied IHC images document human tissue only (catalog reactivity; A07303-1 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P16298 (PP2BB_HUMAN, Serine/threonine-protein phosphatase 2B catalytic subunit beta isoform).
- Human Protein Atlas. PPP3CB tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. PPP3CB subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm and calyx. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. PPP3CB antibody validation summary (2 antibodies).
- Significant role of PPP3CB in malignant gliomas development, prognosis and potential therapeutic application-a study based on comprehensive bioinformatics, cell experiments and immunohistochemistry analyses. Biochemistry and biophysics reports 2024 — PMC10733678.
- A novel interaction between ATOH8 and PPP3CB. Histochemistry and cell biology 2016 — PMC4710663.
- The role of phosphoprotein phosphatases catalytic subunit genes in pancreatic cancer. Bioscience reports 2021 — PMC7785039.
- Calcineurin A beta deficiency ameliorates HFD-induced hypothalamic astrocytosis in mice. Journal of neuroinflammation 2018 — PMC5806488.
- PubMed PMID:2556704 — UniProt-cited evidence.
- PubMed PMID:1848109 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP3CB (Serine/threonine-protein phosphatase 2B catalytic subunit beta isoform). Expected localisation Cytoplasmic (UniProt); extracellular signal in some tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A07303-1). Validated IHC antibody: A07303-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Glandular cells); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human rectum adenocarcinoma tissue; fixative not specified (datasheet A07303-1); 2. Fixation: Image fixative and duration unreported (datasheet A07303-1); 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 A07303-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A07303-1); 8. Primary antibody: Rabbit anti-PPP3CB, 2-5 μg/ml (datasheet A07303-1); 9. Primary incubation: Overnight at 4 °C (datasheet A07303-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07303-1); Caveat: Tissue staining has low concordance with RNA data (HPA tissue IHC); Isoform / epitope: Four isoforms; check epitope coverage (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt P16298, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A07303-1, PMC10733678, PMC7785039. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP3CB/immunohistochemistry-ihc-guide.