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PPP1R14A / Protein phosphatase 1 regulatory subunit 14A · IHC design guide
Design Immunohistochemistry for PPP1R14A
Plan PPP1R14A chromogenic IHC-P with the catalog antibody at 0.5–1 μg/mL (datasheet A05752-1). Compare high staining in lung alveolar type II cells with undetected staining in adipocytes, and assess cytoplasmic and nuclear signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP1R14A 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 and nuclear tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining across tissues and vessels (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A05752-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium agreement with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage unknown (UniProt) |
Section 1
Recommended PPP1R14A IHC & IF Protocols
The catalog antibody protocol (datasheet A05752-1) is accompanied by published PPP1R14A IHC examples in human penile tissue (PMC13097017) and colon tissue (PMC11632118).
| Sample | Paraffin-embedded mouse testis tissues; fixative not specified (datasheet A05752-1) |
| Fixation | Image fixative and duration unreported (datasheet A05752-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A05752-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05752-1) |
| Primary antibody | Rabbit anti-PPP1R14A, 0.5-1μg/ml (datasheet A05752-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05752-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05752-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PPP1R14A-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues and in blood vessles. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for the catalog antibody (datasheet A05752-1); neither cited article excerpt specifies retrieval conditions (PMC13097017; PMC11632118).
Section 2
What Is the Expected PPP1R14A Staining Pattern?
PPP1R14A is annotated as cytoplasmic and has no transmembrane segment (UniProt Q96A00). In paraffin IHC, HPA reports cytoplasmic and nuclear staining across several tissues, including blood vessels; high staining occurs in adrenal glandular cells, brain glial cells and lung alveolar type II cells (HPA tissue IHC). HPA rates the tissue profile Enhanced, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Distinct staining in adrenal glandular cells, brain glial cells or lung alveolar type II cells. | These are reported high-staining cell populations (HPA tissue IHC). Score the identified cells and their compartments; a positive area elsewhere in the section does not establish staining in the intended population (general IHC practice). |
| Cytoplasmic staining, with nuclear staining in some cells. | Both compartments occur in the reported tissue profile (HPA tissue IHC). UniProt annotates cytoplasmic localization (UniProt Q96A00); nuclear signal in a tissue section therefore needs evaluation alongside cell identity and controls. |
| Predominantly extracellular or sharply cell-surface staining, or staining across unrelated cell types. | The reported tissue pattern is cytoplasmic and nuclear (HPA tissue IHC), and no transmembrane segment is annotated (UniProt Q96A00). Check for nonspecific staining, cross-reactivity or detection artefact before assigning these patterns to PPP1R14A (general IHC practice). |
| Diffuse chromogen over tissue and spaces between cells, with poor cell boundaries. | This does not resolve the reported positive cell populations (HPA tissue IHC). Background from detection reagents or incomplete blocking can obscure interpretation; compare a negative reagent control and inspect the counterstain (general IHC practice). |
| No signal in a reported high-staining cell population. | A negative section alone cannot distinguish a technical failure from variation in the specimen or antibody performance (general IHC practice). Recheck the identity of the expected cells; HPA reports high staining in specific cell populations, not every cell in their tissues (HPA tissue IHC). |
💡Expected PPP1R14A appearanceCall a result positive when identifiable reported high-staining cells show resolved cytoplasmic staining, with possible nuclear staining (HPA tissue IHC); diffuse extracellular or indiscriminate staining warrants investigation (general IHC practice).
| Tissue and cell choice | Adrenal glandular cells, brain glial cells and lung alveolar type II cells are reported High; adipocytes in adipose tissue and heart cardiomyocytes are Not detected (HPA tissue IHC). Judge each named cell population, not its whole tissue. |
| Compartment and topology | UniProt annotates cytoplasm and no transmembrane segment (UniProt Q96A00). HPA tissue IHC also reports nuclear staining (HPA tissue IHC); this difference makes cell-resolved assessment useful and does not establish a membrane pattern. |
| Isoforms and processing | UniProt lists two isoforms: isoform 1 in aorta and testis, and isoform 2 in aorta; it annotates no signal peptide or propeptide (UniProt Q96A00). The supplied IHC evidence does not assign staining to either isoform (HPA tissue IHC). |
| Phosphorylation | PPP1R14A has annotated phosphorylation sites, and phosphorylation markedly increases its inhibitory activity (UniProt Q96A00). Total-protein IHC staining does not establish phosphorylation state or inhibitory activity without a separately validated state-specific assay (general IHC practice). |
| Antibody evidence | HPA lists HPA042097 and CAB005044 as IHC Enhanced, and CAB009803 as IHC Supported (HPA antibodies). Apply those ratings to the listed antibodies; the tissue profile still has medium staining–RNA agreement and awaits external verification (HPA tissue IHC). |
| IF/ICC Q&A: Is nuclear signal plausible? | Yes. HPA ICC-IF reports mainly nucleoplasmic localization, with additional nuclear bodies (HPA subcellular). This cell-image result can inform interpretation, while the paraffin IHC tissue profile reports both cytoplasmic and nuclear staining (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported high-staining population appears entirely negative. | The expected cell type may be absent from the section, or the staining run may have failed (general IHC practice); HPA levels describe specific cell populations (HPA tissue IHC). | Confirm cell identity on the counterstain, then review the IHC-validated antibody and run controls. Review retrieval and detection settings as general IHC checks; PPP1R14A-specific fixation sensitivity is unreported in these sources. |
| Chromogen covers the section without clear cell boundaries. | Diffuse background may arise from detection chemistry or nonspecific reagent binding (general IHC practice). It cannot be scored against HPA's cell-specific pattern (HPA tissue IHC). | Compare the negative reagent control, check blocking and washes, and reassess the IHC-validated antibody's working conditions (general IHC practice). Score only staining that can be assigned to intact cells. |
| Staining appears mainly extracellular or confined to cell surfaces. | That distribution departs from the reported cytoplasmic and nuclear tissue pattern (HPA tissue IHC) and the absence of a transmembrane segment (UniProt Q96A00). | Check morphology and counterstain, inspect negative controls, and repeat with an independently validated IHC antibody if available (general IHC practice). Treat the unusual compartment as unresolved until it reproduces. |
| Cells listed as Not detected are strongly stained. | The cell type may be misidentified, or staining may reflect cross-reactivity or endogenous detection activity (general IHC practice). HPA lists adipocytes in adipose tissue and heart cardiomyocytes as Not detected (HPA tissue IHC). | Verify the precise cell type and compare a negative reagent control. If using enzyme-based detection, check the relevant endogenous enzyme block (general IHC practice); avoid judging an entire tissue by one negative cell category. |
| Positive cells show mixed cytoplasmic and nuclear intensity. | Both compartments are reported in tissue IHC (HPA tissue IHC), while HPA ICC-IF emphasizes the nucleoplasm (HPA subcellular); compartment intensity alone does not settle specificity. | Score cytoplasmic and nuclear staining separately in identified cells and compare run controls (general IHC practice). Record the antibody used because HPA validation ratings differ by antibody (HPA antibodies). |
| A faint signal is seen in a reported low-staining population. | HPA reports Low staining in bronchial respiratory epithelial cells and urinary bladder urothelial cells (HPA tissue IHC). Weak signal can be difficult to separate from background (general IHC practice). | Compare cells with nearby background and the negative reagent control, then record intensity with cell identity (general IHC practice). Use a reported high-staining population to check run performance (HPA tissue IHC). |
Experiment evidence
IHC & IF images showing PPP1R14A staining
A05752-1 · IHCIHC analysis of CPI17 alpha using anti-CPI17 alpha antibody (A05752-1). CPI17 alpha was detected in paraffin-embedded section of mouse testi
A05752-1 · IHCIHC analysis of CPI17 alpha using anti-CPI17 alpha antibody (A05752-1). CPI17 alpha was detected in paraffin-embedded section of rat lung ti
A05752-1 · IFIF analysis of CPI17 alpha using anti-CPI17 alpha antibody (A05752-1). CPI17 alpha was detected in immunocytochemical section of U20S cells.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000167641-PPP1R14A/tissue/cerebral+cortex#imid_2018720
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000167641-PPP1R14A/tissue/adrenal+gland#imid_3714992
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000167641-PPP1R14A/tissue/colon#imid_2018654
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000167641-PPP1R14A/tissue/liver#imid_2018655
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB005044; green = PPP1R14A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB005044; green = PPP1R14A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP1R14A IHC & IF
🧪Run adrenal gland first and score its glandular cells, which are High for PPP1R14A (HPA: High in adrenal gland glandular cells). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the adrenal slide, treat non-glandular cells as internal background comparators rather than assuming they are target-negative (HPA: High designation applies to glandular cells).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP1R14A in A-431, U2OS, A-549, CACO-2, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit isotype control for the rabbit catalog antibody (caption: rabbit primary); a PPP1R14A knockout specimen, if available, provides a biological specificity control. Quench endogenous peroxidase and check endogenous biotin background on adrenal sections because the reported IHC detection uses biotin–streptavidin and DAB (caption: SABC with DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A05752-1 paraffin-section caption does not state the fixative (caption: fixative not stated). The reported IHC procedure uses heat retrieval in citrate buffer at pH 6 for 20 minutes; whether retrieval is required for other preparations and whether frozen sections or IF are easier is unreported (caption: citrate retrieval). No adrenal-specific artefact is established by the supplied evidence; assess background with the controls above (HPA: High in adrenal gland glandular cells).
HPA tissue IHC evidence for PPP1R14A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Granular cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PPP1R14A IHC Tips
Troubleshooting PPP1R14A staining in paraffin sections requires attention to retrieval, compartment, cell type and the limits of antibody validation.
How should I adjust retrieval when PPP1R14A staining is weak?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes on paraffin sections (datasheet A05752-1). The reported mouse testis IHC used that condition before overnight incubation with 1 μg/ml primary antibody at 4°C (datasheet A05752-1). If staining remains weak, compare a small matched section series using a longer citrate heating interval or EDTA at pH 9 as fallback conditions (standard IHC practice). Keep detection and exposure to chromogen constant, and assess signal against tissue morphology and a no-primary control so increased background is not mistaken for improved retrieval (standard IHC practice).
Could fixation explain inconsistent PPP1R14A staining between paraffin blocks?
The selected mouse testis caption identifies paraffin sections but does not state the fixative, so target-specific fixation sensitivity is unknown (datasheet A05752-1). Record fixative, fixation duration and tissue processing for each block before comparing stain intensity; variation in these steps can alter IHC performance generally (standard IHC practice). For a controlled pilot, process matched samples with a documented condition such as 10% neutral buffered formalin for 24 hours, then apply the same citrate pH 6 retrieval (standard IHC practice; datasheet A05752-1). Treat any difference as an empirical finding for those samples, rather than inferring PPP1R14A fixation sensitivity from tissue expression or protein features (standard IHC practice).
Should PPP1R14A signal be cytoplasmic or nuclear in tissue sections?
UniProt lists PPP1R14A as cytoplasmic and identifies no transmembrane segment (UniProt Q96A00 subcellular location and topology). HPA tissue IHC describes cytoplasmic and nuclear expression, while its cell imaging supports nucleoplasmic localisation with additional nuclear bodies (HPA tissue IHC; HPA subcellular). Score cytoplasmic and nuclear chromogen separately within defined cell types instead of calling either compartment universally nonspecific; include a nuclear counterstain to resolve the boundary (HPA tissue IHC; standard IHC practice). If the nuclear pattern varies sharply across adjacent sections, review retrieval, background controls and tissue preservation before assigning a biological explanation (standard IHC practice).
Can this IHC stain distinguish PPP1R14A isoforms or phosphorylation states?
PPP1R14A has 2 annotated isoforms: isoform 1 is reported in aorta and testis, and isoform 2 in aorta (UniProt Q96A00 isoforms and tissue specificity). Its annotated modifications include PKC-associated phosphothreonine at residue 38 and phosphoserines at residues 26, 128, 134 and 136 (UniProt Q96A00 modified residues). The supplied IHC caption gives no epitope map or evidence of isoform or phospho-state selectivity for A05752-1 (datasheet A05752-1). Interpret chromogenic intensity as antibody staining until epitope mapping or suitable isoform and phospho-specific controls establish what the assay distinguishes (standard IHC practice).
How should I investigate a PPP1R14A pattern by multiplex IF?
On the separate IF/ICC guide, pair PPP1R14A with a marker for the cell population under study, such as an established smooth muscle marker when examining aorta (UniProt Q96A00 tissue specificity and function; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel for the weaker signal after checking tissue autofluorescence with an unstained control (standard IF practice). Because reported PPP1R14A locations include cytoplasm and nucleoplasm, use controlled permeabilisation for intracellular access and compare it with a milder condition if morphology suffers (UniProt Q96A00 subcellular location; HPA subcellular; standard IF practice). Include single-stain controls to identify bleed-through before interpreting apparent colocalisation (standard IF practice).
What should I check when PPP1R14A DAB staining is diffuse?
The reported IHC used 10% goat serum block, 1 μg/ml rabbit primary overnight at 4°C, and biotin-based detection with DAB (datasheet A05752-1). Run a no-primary control and inspect it for endogenous peroxidase activity or biotin-related staining before assigning diffuse brown signal to PPP1R14A (standard IHC practice). Apply a peroxidase block and, if background persists with this detection system, compare a biotin-free detection method on matched sections (standard IHC practice). Check whether background tracks tissue edges, damaged areas or every cell equally, then titrate primary concentration and washing conditions while preserving a positive reference section (standard IHC practice).
How can I score PPP1R14A IHC across heterogeneous tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical region and cell type before scoring, since HPA reports different staining levels across tissues and cell populations (HPA tissue IHC). For each compartment, record the percentage of positive cells and intensity on a 0–3 scale; an H-score can combine them over a 0–300 range (standard IHC practice). When counting discrete positive cells, report density per mm² of viable, evaluable tissue and exclude folds, necrosis and empty space (standard IHC practice). Normalize comparisons to the same cell population, section area, staining run and reference control, and keep cytoplasmic and nuclear scores separate (HPA tissue IHC; standard IHC practice).
How do I distinguish plausible PPP1R14A staining from artefact?
Expect a pattern that is reproducible in the same cell population across intact sections and supported by a clean no-primary control (standard IHC practice). HPA reports high staining in glial cells in several brain regions and in adrenal glandular cells, but its tissue IHC has only medium agreement with RNA data and awaits external verification (HPA tissue IHC). Compare cytoplasmic and nuclear signal independently, because UniProt lists cytoplasm while HPA also reports nuclear staining; neither location alone proves specificity (UniProt Q96A00 subcellular location; HPA tissue IHC). Discount edge-only staining, necrotic deposits and signal reproduced by the no-primary control, including endogenous enzyme activity (standard IHC practice).
Boster reagents
Best PPP1R14A / Protein phosphatase 1 regulatory subunit 14A IHC Antibodies
A05752-1 has IHC images from paraffin sections of mouse testis, rat lung, and human intestinal and lung cancers (IHC image captions), plus IF data from U20S cells (IF image caption).
Real IHC data
Anti-CPI17 alpha/PPP1R14A Antibody ®
Cat # A05752-1
A05752-1 is listed for IHC and IF/ICC, with human, mouse, and rat reactivity (catalog applications/reactivity). Its IHC captions show the paraffin sections listed above, while its IF caption shows U20S cells (catalog image captions).
Which to pick: Choose A05752-1 for paraffin-section chromogenic IHC; its captions document citrate retrieval and DAB detection, with a listed IHC concentration of 0.5–1 μg/ml (IHC image captions; catalog dilution). For IF/ICC, A05752-1 has an IF image from U20S cells and a listed concentration of 2 μg/ml (IF image caption; catalog dilution). It is listed as reactive with human, mouse, and rat; the host is rabbit, clonality is unreported, and the IHC captions do not report the fixative (catalog reactivity/host/clone; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96A00 (PP14A_HUMAN, Protein phosphatase 1 regulatory subunit 14A).
- Human Protein Atlas. PPP1R14A tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PPP1R14A subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies..
- Human Protein Atlas. PPP1R14A antibody validation summary (4 antibodies).
- PPP1R14A in male erectile dysfunction: key role in smooth muscle cells. Sexual medicine 2026 — PMC13097017.
- Role of eosinophil counts in mediating the association between asthma and colon cancer. Clinical and translational allergy 2024 — PMC11632118.
- PubMed PMID:11467857 — UniProt-cited evidence.
- PubMed PMID:12974676 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP1R14A (Protein phosphatase 1 regulatory subunit 14A). Expected localisation Cytoplasmic and nuclear tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A05752-1). Validated IHC antibody: A05752-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse testis tissues; fixative not specified (datasheet A05752-1); 2. Fixation: Image fixative and duration unreported (datasheet A05752-1); 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: Citrate pH 6 , 20 min (datasheet A05752-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05752-1); 8. Primary antibody: Rabbit anti-PPP1R14A, 0.5-1μg/ml (datasheet A05752-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05752-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium agreement with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q96A00, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A05752-1, PMC13097017, PMC11632118. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP1R14A/immunohistochemistry-ihc-guide.