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PPP1R12A / Protein phosphatase 1 regulatory subunit 12A · IHC design guide
Design Immunohistochemistry for PPP1R12A
Plan PPP1R12A staining in paraffin sections around the cytoplasmic and membranous pattern seen in most cells (HPA tissue IHC). Breast glandular cells stain strongly, while adipocytes are reported as undetected (HPA tissue IHC); start the catalog antibody at 0.5–1 µg/mL (datasheet: PB9737).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PPP1R12A 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 membranous (HPA tissue IHC) | |
| Staining pattern | Most cells show cytoplasmic and membranous staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9737) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes may lack staining despite broad tissue expression (HPA tissue IHC) | |
| Regulation | No stimulus-dependent regulation specified (UniProt) | |
| Isoform / epitope | Five isoforms; confirm antibody epitope coverage (UniProt) |
Section 1
Recommended PPP1R12A IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published PPP1R12A staining methods for tumor sections (PMC8210562) and pancreatic tissues (PMC4843125).
| Sample | Paraffin-embedded Human Glioma tissue; fixative not specified (datasheet PB9737) |
| Fixation | Image fixative and duration unreported (datasheet PB9737); 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 PB9737) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9737) |
| Primary antibody | Rabbit anti-PPP1R12A, 0.5-1μg/ml (datasheet PB9737) |
| Primary incubation | Overnight at 4 °C (datasheet PB9737) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9737) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PPP1R12A-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6, as specified by the catalog datasheet and used in the tumor-section protocol (datasheet: PB9737; PMC8210562).
Section 2
What Is the Expected PPP1R12A Staining Pattern?
PPP1R12A is a cytoplasmic protein associated with stress fibers and actomyosin filaments; it has no transmembrane segment (UniProt O14974). In paraffin-section IHC, expect cytoplasmic staining, sometimes with a membranous appearance, across many cell types (HPA tissue IHC). HPA rates the tissue staining Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic chromogen in breast or endometrial glandular cells, with some apparent membrane staining. | This fits the reported cytoplasmic and membranous pattern; both named glandular populations stain High (HPA tissue IHC). Judge the signal within cells against adjacent background and a negative reagent control (general IHC practice). |
| Strong, widespread nuclear staining replaces the cytoplasmic pattern. | Treat this as a possible artefact and review the controls and staining conditions: cytoplasm and stress fibers are the UniProt locations (UniProt O14974). HPA reports uncertain nucleolar localization in ICC-IF, which does not establish widespread nuclear IHC staining (HPA subcellular). |
| Adipocytes stain as strongly as the selected positive tissue. | Adipocytes are reported Not detected, so this result warrants a specificity check (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible; compare a negative reagent control and an independently validated antibody before assigning the signal to PPP1R12A (general IHC practice). |
| Chromogen covers tissue spaces or obscures cell boundaries. | Diffuse background cannot establish a PPP1R12A-positive cell or compartment (general IHC practice). Check the negative reagent control, blocking, washes, and detection reagent; retain the tissue context because HPA reports expression in most cells (HPA tissue IHC). |
| No signal appears in a breast glandular-cell or bronchial epithelial-cell section. | Both populations are reported High, so first check tissue preservation and the run controls (HPA tissue IHC; general IHC practice). A failed run, unsuitable retrieval, or antibody conditions could explain absent signal; HPA supplies no target-specific fixation-sensitivity claim. |
💡Expected PPP1R12A appearanceCall a positive result when cellular cytoplasmic chromogen is clear in an HPA High population, such as breast glandular cells; diffuse background or strong staining of adipocytes is suspect (HPA tissue IHC; general IHC practice).
| Cellular location | Cytoplasm and stress fibers are reported locations, including association with actomyosin filaments (UniProt O14974). HPA tissue IHC also describes membranous expression; assess that appearance alongside cytoplasmic staining (HPA tissue IHC). |
| Tissue and cell choice | Breast glandular cells, bronchial respiratory epithelium, and cortical neurons are High; adipocytes are Not detected (HPA tissue IHC). Low tissue specificity means a positive result elsewhere is not automatically unexpected (HPA tissue IHC). |
| Antibody evidence | HPA071956 is Approved for IHC, while HPA041296 has no stated IHC status (HPA antibodies). HPA's overall Approved tissue rating has medium RNA–staining consistency; neither statement validates an unspecified catalog antibody (HPA tissue IHC; HPA antibodies). |
| IF/ICC Q: What location should a separate IF guide assess? | A: Mainly cytosol and actin filaments; HPA marks both Supported. Nucleoli and plasma membrane are additional, Uncertain locations (HPA subcellular). This ICC-IF evidence is a localization comparison, not an IHC protocol or an IHC validation claim. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive tissue gives no chromogen. | The assay may have failed, or retrieval and antibody conditions may be unsuitable (general IHC practice). HPA lists breast glandular cells as High (HPA tissue IHC). | Check run controls and section integrity, then review the IHC-validated antibody's documented retrieval and dilution conditions (general IHC practice). Do not infer PPP1R12A-specific fixation sensitivity from the HPA tissue score. |
| Adipocytes show strong cellular staining. | This conflicts with the reported Not detected adipocyte result (HPA tissue IHC). Cross-reactivity or endogenous detection activity may contribute (general IHC practice). | Compare a negative reagent control and an HPA High cell population on the same run; investigate detection activity and antibody specificity before scoring adipocytes positive (HPA tissue IHC; general IHC practice). |
| Most nuclei are dark while cytoplasm is weak. | A predominantly nuclear result does not match UniProt's cytoplasmic and stress-fiber locations (UniProt O14974). HPA's additional nucleolar ICC-IF signal is Uncertain (HPA subcellular). | Inspect controls, counterstain, and chromogen distribution; repeat under documented IHC conditions if needed (general IHC practice). Do not use uncertain ICC-IF nucleolar staining to validate broad nuclear IHC. |
| The whole section has hazy chromogen. | Background from detection reagents, inadequate blocking, or insufficient washing can obscure cellular localization (general IHC practice). HPA's broad expression pattern alone cannot identify a diffuse haze as specific staining (HPA tissue IHC). | Review the negative reagent control, blocking, washes, and chromogen development (general IHC practice). Score only discernible cellular signal in the expected context (HPA tissue IHC). |
| Membrane-like staining is the only prominent signal. | HPA describes membranous tissue staining, but plasma-membrane localization is Uncertain in ICC-IF (HPA tissue IHC; HPA subcellular). The two observations do not establish a membrane-only target pattern. | Look for accompanying cytoplasmic staining and compare a High reference population and assay controls (HPA tissue IHC; general IHC practice). Report the observed distribution without claiming confirmed plasma-membrane localization. |
| Signal varies markedly between sections in one run. | Section quality, retrieval, and reagent handling can alter IHC signal (general IHC practice). HPA's Approved rating still notes medium consistency with RNA expression (HPA tissue IHC). | Compare run controls and the same HPA High cell population across sections; review processing and assay records before interpreting variation as biology (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PPP1R12A staining
PB9737 · IHCIHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737). PPP1R12A was detected in paraffin-embedded section of Human Glioma Tissue. H
PB9737 · IHCIHC analysis of PPP1R12A using anti-PPP1R12A antibody (PB9737). PPP1R12A was detected in paraffin-embedded section of Human Glioma Tissue. H
M01743-1 · IHCHuman placenta was stained with anti-MYPT1 rabbit antibody
PA1681 · IFIF analysis of PPP1R12A using anti-PPP1R12A antibody (PA1681). PPP1R12A was detected in immunocytochemical section of SiHa cells. Enzyme ant
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000058272-PPP1R12A/tissue/cerebral+cortex#imid_20166373
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000058272-PPP1R12A/tissue/colon#imid_20165435
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000058272-PPP1R12A/tissue/liver#imid_20165452
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000058272-PPP1R12A/tissue/kidney#imid_20165437
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041296; green = PPP1R12A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041296; green = PPP1R12A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PPP1R12A IHC & IF
🧪Run appendix first and assess PPP1R12A staining in its lymphoid tissue (HPA: High in appendix lymphoid tissue). Use adipose tissue as the negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); HPA does not identify a confirmed negative cell population within appendix, so treat other cells on that slide as internal negatives only if independently validated.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PPP1R12A in A-431, U-251MG, U2OS, RT-4, SiHa, RPTEC/TERT1, hTERT-RPE1 (serum starved), NIH 3T3, with annotated localisation: Actin filaments (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a concentration-matched rabbit isotype control for the rabbit catalog antibody, and a PPP1R12A knockout control or peptide block if a suitable blocking peptide is available (caption: rabbit anti-PPP1R12A; standard IHC control practice). For appendix, quench endogenous peroxidase and check for endogenous biotin background when using the reported avidin–biotin/DAB detection (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9737 paraffin-section caption does not state the fixative (caption: fixative not stated). The caption reports heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish that retrieval is required under other processing conditions (caption: citrate retrieval). There is no supplied evidence that frozen sections are easier; IF may help assess cytosol and actin-filament localization (HPA: supported main locations).
HPA tissue IHC evidence for PPP1R12A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PPP1R12A IHC Tips
Troubleshoot PPP1R12A chromogenic IHC in paraffin sections using the PB9737 tissue caption, protein localisation data, and matched staining controls.
What retrieval should I try first when PPP1R12A staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet PB9737 tissue-IHC caption). The reported paraffin-section image used this retrieval before overnight incubation with 1 μg/mL catalog antibody at 4°C (datasheet PB9737 tissue-IHC caption). If staining remains weak, compare a longer citrate retrieval with the original condition on adjacent sections, while holding antibody concentration and detection constant (standard IHC practice). Check tissue integrity and background as well as signal: excessive heating can damage morphology and make cytoplasmic staining harder to assign to individual cells (standard IHC practice).
Could fixation explain weak or patchy PPP1R12A staining?
The PB9737 tissue-IHC caption identifies a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet PB9737 tissue-IHC caption). Record the fixative, fixation duration, tissue thickness, and processing history for each specimen before comparing staining intensity (standard IHC practice). On matched sections, keep citrate retrieval at pH 6 for 20 minutes and the catalog antibody at 1 μg/mL while examining whether staining tracks processing differences (datasheet PB9737 tissue-IHC caption; standard IHC practice). Evaluate morphology alongside staining, because uneven preservation can make cytoplasmic signal appear patchy without establishing a PPP1R12A-specific fixation effect (standard IHC practice).
Where should convincing PPP1R12A staining appear in tissue sections?
Expect predominantly cytoplasmic staining, potentially following contractile structures, because PPP1R12A is reported in cytoplasm, stress fibers, and along actomyosin filaments (UniProt O14974 localisation). Tissue IHC also reports cytoplasmic and membranous expression in most cells, while its staining-to-RNA consistency is medium (HPA tissue IHC). Interpret apparent membrane outlines cautiously: supported cell-imaging locations are cytosol and actin filaments, whereas plasma membrane localisation is uncertain (HPA subcellular). Compare staining with cell boundaries and a morphology-preserving counterstain; a sharply nuclear-only pattern warrants review of controls and staining conditions (HPA subcellular; standard IHC practice).
Could an isoform or modified epitope change the staining pattern?
PPP1R12A has 5 annotated isoforms and multiple modified residues, including hydroxyasparagines and phosphoserines (UniProt O14974 isoforms; UniProt O14974 modified residues). The supplied PB9737 caption does not identify the antibody epitope, so it cannot establish which isoforms or modification states contribute to staining (datasheet PB9737 tissue-IHC caption). If specimens differ unexpectedly, check the catalog antibody’s immunogen or epitope documentation before attributing the difference to alternative splicing or phosphorylation (standard IHC practice). Compare adjacent sections under identical retrieval and detection conditions, then seek an independent epitope or orthogonal assay before making an isoform-specific claim (standard IHC practice).
How can IF help assess an ambiguous chromogenic IHC pattern?
Use IF as a separate localisation check, since the PB9737 evidence supplied here documents chromogenic tissue IHC rather than an IF procedure (datasheet PB9737 tissue-IHC caption). Multiplex PPP1R12A with a validated marker for the expected cell type in the specimen, and assess whether signals occupy the same cells (standard IF practice). Choose spectrally separated fluorophores, favouring a far-red channel when tissue autofluorescence obscures shorter-wavelength signal, and include single-channel controls (standard IF practice). Because PPP1R12A has no transmembrane segment and its supported locations include cytosol and actin filaments, assess intracellular access with appropriate permeabilisation for the IF preparation (UniProt O14974 topology; HPA subcellular; standard IF practice).
How do I distinguish diffuse PPP1R12A staining from detection background?
The reported PB9737 workflow uses biotinylated secondary antibody, streptavidin-biotin detection, and DAB, so inspect controls for background from those detection steps (datasheet PB9737 tissue-IHC caption; standard IHC practice). Run a no-primary section through the same secondary and chromogen steps, and compare it with a section stained using the complete workflow (standard IHC practice). If the no-primary section develops color, review endogenous biotin, endogenous peroxidase blocking, secondary-antibody binding, and DAB development before changing the primary antibody (standard IHC practice). Keep the reported 10% goat-serum block and 1 μg/mL primary concentration as reference conditions when testing one change at a time (datasheet PB9737 tissue-IHC caption; standard IHC practice).
What is a defensible way to quantify PPP1R12A IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, since HPA reports PPP1R12A across many cell types and variable staining levels (HPA tissue IHC). For cellular staining, report the percentage of positive cells and an H-score from 0–300, based on percentages at 0, 1, 2, and 3 intensity levels (standard IHC practice). If measuring stained area, report positive area per mm² and normalise it to viable tissue area or the counted target-cell population (standard IHC practice). Apply one threshold and imaging setup across batches, exclude artefactual regions by predefined rules, and retain representative controls for review (standard IHC practice).
Which findings would make a PPP1R12A-positive result unconvincing?
A convincing result should fit predominantly cytoplasmic PPP1R12A localisation and be interpretable within identifiable cells (UniProt O14974 localisation; HPA subcellular). HPA reports high staining in several populations, including breast glandular cells and cerebral-cortex neurons, but reports adipocytes as not detected; use only tissue-matched expectations (HPA tissue IHC). Treat nuclear-only staining, section-edge enhancement, staining confined to necrotic areas, or color in a no-primary control as reasons to investigate artefact (HPA subcellular; standard IHC practice). In the reported biotin-based DAB workflow, check endogenous enzyme activity and detection background before assigning such color to PPP1R12A (datasheet PB9737 tissue-IHC caption; standard IHC practice).
Boster reagents
Best PPP1R12A / Protein phosphatase 1 regulatory subunit 12A IHC Antibodies
Real IHC images cover human glioma paraffin sections (PB9737 caption) and human placenta (M01743-1 caption); an IF/ICC image covers SiHa cells (PA1681 caption).
Real IHC data
Anti-Myosin Phosphatase/PPP1R12A Antibody ®
Cat # PB9737
Real IHC data
Anti-MYPT1 Rabbit Monoclonal Antibody
Cat # M01743-1
Real IF data
Anti-Myosin Phosphatase/PPP1R12A Antibody ®
Cat # PA1681
PB9737 has an IHC image of human glioma paraffin sections and lists IHC and IF applications (PB9737 caption; catalog applications); M01743-1 has a human placenta IHC image and lists IHC and IF applications (M01743-1 caption; catalog applications). PA1681 has an IF/ICC image of SiHa cells and lists IF and ICC applications (PA1681 caption; catalog applications).
Which to pick: For tissue IHC, choose PB9737 for its human glioma paraffin-section image (PB9737 caption), or M01743-1 for its human placenta IHC image and monoclonal format (M01743-1 caption; catalog); neither caption reports a fixative (PB9737 caption; M01743-1 caption). For IF/ICC, PA1681 has a SiHa cell image but no listed IHC application (PA1681 caption; catalog applications). For work across species, PB9737 lists human, monkey, mouse and rat reactivity, while its pictured tissue IHC result is human (PB9737 catalog reactivity; PB9737 caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14974 (MYPT1_HUMAN, Protein phosphatase 1 regulatory subunit 12A).
- Human Protein Atlas. PPP1R12A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PPP1R12A subcellular location (ICC-IF): Mainly localized to the cytosol and actin filaments. In addition localized to the nucleoli and plasma membrane..
- Human Protein Atlas. PPP1R12A antibody validation summary (2 antibodies).
- MicroRNA-455-5p Contributes to Cholangiocarcinoma Growth and Mediates Galangin's Anti-Tumor Effects. Journal of Cancer 2021 — PMC8210562.
- Identification and Validation of a PPP1R12A-Related Five-Gene Signature Associated With Metabolism to Predict the Prognosis of Patients With Prostate Cancer. Frontiers in genetics 2021 — PMC8414655.
- Trans-Synaptic Signaling through the Glutamate Receptor Delta-1 Mediates Inhibitory Synapse Formation in Cortical Pyramidal Neurons. Neuron 2019 — PMC6926483.
- Using Quantitative Seroproteomics to Identify Antibody Biomarkers in Pancreatic Cancer. Cancer immunology research 2016 — PMC4843125.
- PubMed PMID:9286714 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PPP1R12A (Protein phosphatase 1 regulatory subunit 12A). Expected localisation Cytoplasmic and membranous (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9737). Validated IHC antibody: PB9737 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human Glioma tissue; fixative not specified (datasheet PB9737); 2. Fixation: Image fixative and duration unreported (datasheet PB9737); 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 PB9737); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9737); 8. Primary antibody: Rabbit anti-PPP1R12A, 0.5-1μg/ml (datasheet PB9737); 9. Primary incubation: Overnight at 4 °C (datasheet PB9737). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Adipocytes may lack staining despite broad tissue expression (HPA tissue IHC); Isoform / epitope: Five isoforms; confirm antibody epitope coverage (UniProt); Regulation: No stimulus-dependent regulation specified (UniProt). Evidence: UniProt O14974, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol and actin filaments. In addition…, Datasheet PB9737, PMC8210562, PMC4843125. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PPP1R12A/immunohistochemistry-ihc-guide.