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PRKCE / Protein kinase C epsilon type · IHC design guide
Design Immunohistochemistry for PRKCE
Plan chromogenic PRKCE IHC in paraffin sections around cytoplasmic and membranous staining, with high signal in cerebral cortical neurons and kidney distal tubules (HPA tissue IHC). The catalog antibody lists an IHC concentration of 0.5–1 μg/ml (datasheet: PB9320); compare stained cells with documented negative tissue populations (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRKCE 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | High in cortical neurons and kidney distal tubules (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9320) | |
| Positive control | Cerebral cortex+3 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9320) | |
| Caveat | HGF can shift epithelial PRKCE to the plasma membrane (UniProt) | |
| Regulation | HGF-dependent membrane redistribution (UniProt) | |
| Isoform / epitope | No listed isoforms or cleavage; no extracellular domain (UniProt) |
Section 1
Recommended PRKCE IHC & IF Protocols
Compare the catalog antibody's IHC-P protocol (datasheet: PB9320) with three published PRKCE IHC protocols (PMC7533283; PMC9293992; PMC5520703).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PB9320); 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 PB9320); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9320) |
| Primary antibody | Rabbit anti-PRKCE, 0.5-1μg/ml (datasheet PB9320) |
| Primary incubation | Overnight at 4 °C (datasheet PB9320) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9320) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRKCE-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in few tissues, most abundant expression in CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: PB9320). The published protocols specify citric acid or microwave retrieval where reported (PMC7533283; PMC9293992).
Section 2
What Is the Expected PRKCE Staining Pattern?
PRKCE is expected mainly in cytoplasmic and membranous compartments on tissue IHC, with the strongest listed staining in cerebral cortical neurons and kidney distal tubules (HPA: tissue IHC). HPA rates its tissue staining reliability Enhanced (HPA: reliability). A peripheral signal can fit its membrane localisation, but PRKCE has no transmembrane segment; a purely surface-restricted pattern needs scrutiny (UniProt Q02156: topology and subcellular location).
| Cortical neurons or kidney distal tubules show strong cytoplasmic staining, with some membrane-associated signal (HPA: High; tissue IHC). | This matches the reported cell types and compartments. Compare intensity within the named cells, since adjacent unstained cells do not by themselves contradict the cell-specific HPA observations (HPA: tissue IHC). |
| A section shows mainly nuclear staining, with little cytoplasmic signal (HPA: cytoplasmic and membranous tissue profile). | Treat this as a localisation mismatch and investigate artefact. UniProt also lists the nucleus and reports stimulus-associated nuclear translocation by similarity, so nuclear staining alone cannot establish either an error or activation (UniProt Q02156: subcellular location). |
| Strong staining appears in appendix glandular cells or bone marrow hematopoietic cells (HPA: Not detected in those cells). | This conflicts with the listed HPA observations and warrants a specificity check. Cross-reactivity or endogenous detection activity are possibilities, not proven causes; assess a control lacking primary antibody (HPA: tissue IHC; general IHC practice). |
| Chromogen covers tissue broadly, including cells expected to be unstained (HPA: cell-specific tissue IHC). | Diffuse deposit obscures the reported cell pattern. Check primary-free and detection controls, blocking, washes, and chromogen development before assigning the signal to PRKCE (general IHC practice; HPA: tissue IHC). |
| No signal is visible in cerebral cortical neurons or kidney distal tubules (HPA: High in both cell types). | A failed positive control is plausible. Review tissue identity, antibody suitability, retrieval, detection and counterstain before interpreting a study section as PRKCE-negative (HPA: tissue IHC; general IHC practice). |
💡Expected PRKCE appearanceCall a result positive when cortical neurons or kidney distal tubules show strong, predominantly cytoplasmic staining with possible membrane signal (HPA: High; cytoplasmic and membranous); broad staining of HPA-undetected cell types is suspect (HPA: tissue IHC).
| Cell-specific reference pattern (HPA: tissue IHC) | Cerebral cortical neurons and kidney distal tubules are High; caudate neurons and hippocampal synapses are Medium. Use the named cells when comparing sections, because HPA levels do not describe every cell in each tissue (HPA: tissue IHC). |
| Lower and undetected reference cells (HPA: tissue IHC) | Cerebellar granular-layer cells are Low; appendix glandular cells and bone marrow hematopoietic cells are Not detected. These are cell-level observations, not claims that entire tissue sections lack PRKCE (HPA: tissue IHC). |
| Antibody evidence (HPA: antibody validation) | The tissue profile is rated Enhanced. HPA lists IHC Enhanced for HPA044496 and CAB001948, while HPA008236 has ICC Supported with no IHC status listed. Match validation to the assay being interpreted (HPA: antibodies; reliability). |
| Compartment and stimulus (UniProt Q02156: subcellular location) | UniProt lists cytoplasm, cytoskeleton, cell membrane, perinuclear region and nucleus. It reports HGF-associated plasma-membrane translocation in stimulated epithelial cells; its Golgi and PMA-related nuclear notes are by similarity. The observed context matters (UniProt Q02156). |
| Topology (UniProt Q02156: topology) | PRKCE has no transmembrane segment, signal peptide or propeptide and is listed as one chain spanning residues 1–737. Membrane-associated staining is compatible with its localisation, but topology alone cannot locate the antibody epitope or predict staining after retrieval (UniProt Q02156). |
| IF/ICC Q&A: Is its pattern the same? (HPA: subcellular ICC-IF) | HPA supports cytosol and intermediate filaments, with additional plasma membrane and vesicle localisation in ICC-IF. These cultured-cell observations help assess compartments; they do not prescribe an IF protocol or replace tissue IHC controls (HPA: subcellular ICC-IF; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Cortical neurons or distal tubules are blank (HPA: High in these cells). | The positive control may have failed; the image alone cannot identify which workflow step failed (HPA: tissue IHC; general IHC practice). | Confirm the named cells are present, then check antibody IHC validation, retrieval, primary incubation and detection with a known-positive section (HPA: antibodies and tissue IHC; general IHC practice). |
| Staining is mostly nuclear (HPA: cytoplasmic and membranous tissue profile). | Localisation is discordant with the tissue profile; UniProt also allows context-dependent nuclear localisation (UniProt Q02156: subcellular location). | Compare with a validated positive section and primary-free control; record stimulation context before interpreting nuclear signal as PRKCE (HPA: tissue IHC; UniProt Q02156; general IHC practice). |
| Appendix glandular cells stain strongly (HPA: Not detected). | The result conflicts with HPA's cell-specific observation; nonspecific binding or endogenous detection activity may contribute (HPA: tissue IHC; general IHC practice). | Inspect a primary-free control, review blocking and detection, and compare the same run with a named positive cell population (HPA: tissue IHC; general IHC practice). |
| Deposit is diffuse across cells and extracellular areas (general IHC practice). | Background can arise from nonspecific detection or excessive chromogen development; appearance alone cannot assign a cause (general IHC practice). | Review primary-free and detection controls, blocking, washes and development time; score cells only after background is resolved (general IHC practice). |
| A cell reported Low looks as strong as a High reference cell (HPA: tissue IHC). | Run conditions or image display may distort relative intensity; HPA categories are observations rather than a quantitative assay scale (HPA: tissue IHC; general IHC practice). | Compare matched sections stained and viewed under the same conditions, then describe intensity and cell type rather than forcing a numerical threshold (HPA: tissue IHC; general IHC practice). |
| ICC-IF shows filaments while tissue IHC looks broadly cytoplasmic (HPA: subcellular ICC-IF; tissue IHC). | HPA reports supported intermediate filaments in ICC-IF and a cytoplasmic tissue profile; these are different preparations (HPA: subcellular ICC-IF; tissue IHC). | Interpret each assay against its own HPA reference and validation status; use tissue positive and detection controls for the IHC call (HPA: antibodies and tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PRKCE staining
PB9320 · IHCAnti-PKC epsilon vendor antibody, PB9320, IHC(P) IHC(P): Mouse Intestine Tissue
PB9320 · IHCAnti-PKC epsilon vendor antibody, PB9320, IHC(P) IHC(P): Rat Intestine Tissue
A01151-1 · IFIF analysis of PRKCE4 using anti-PRKCE4 antibody (A01151-1) and anti-Tubulin Alpha antibody (M03989-3). PRKCE4 was detected in immunocytoche
HPA · HippocampusHuman Protein Atlas IHC, Hippocampus. Source: https://www.proteinatlas.org/ENSG00000171132-PRKCE/tissue/hippocampal+formation#imid_1222
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000171132-PRKCE/tissue/cerebral+cortex#imid_20149237
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000171132-PRKCE/tissue/colon#imid_20149163
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000171132-PRKCE/tissue/liver#imid_20149172
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA008236; green = PRKCE, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA008236; green = PRKCE, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRKCE IHC & IF
🧪Run cerebral cortex first; neuronal cells should stain (HPA: High in cerebral-cortex neuronal cells). Use adipose tissue as the negative tissue, focusing on adipocytes (HPA: Not detected in adipocytes); adjacent non-neuronal cells on the cortex slide should be assessed for background staining as an internal comparator, without assuming they are PRKCE-negative.
Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRKCE in Rh30, SK-MEL-30, U2OS, with annotated localisation: Intermediate filaments (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and a biological specificity control using PRKCE-knockout material or immunizing-peptide blocking if available (standard IHC practice). Quench endogenous peroxidase in cerebral-cortex sections for chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected PB9320 intestine IHC(P) caption does not state a fixative (selected-SKU caption: fixative unreported). Retrieval dependency is also unreported, so compare retrieved and unretrieved paraffin sections during optimization (standard IHC practice). Whether frozen sections or IF are easier is unestablished; IF may help assess cytosol and intermediate-filament localization, while cerebral-cortex pigment can complicate signal interpretation (HPA: supported cytosol and intermediate-filament localization; standard IHC/IF practice).
HPA tissue IHC evidence for PRKCE
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. External characterization data supports antibody staining.). 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 |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Kidney | Distal tubules | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Synapses | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PRKCE IHC Tips
Troubleshoot PRKCE staining in paraffin sections by comparing compartment-specific signal with matched controls and documented processing conditions.
How should I optimize retrieval when PRKCE staining is weak in paraffin sections?
Start with heat-mediated EDTA retrieval at pH 8.0 for PRKCE paraffin-section IHC (datasheet PB9320). Keep heating conditions consistent across test and control sections, then compare shorter and longer retrieval on adjacent sections while checking tissue morphology (standard IHC practice). Use neuronal cells in cerebral cortex or distal tubules in kidney as positive reference patterns, and assess cytoplasmic and membranous staining separately (HPA: High in cerebral cortex neuronal cells and kidney distal tubules; HPA: cytoplasmic and membranous tissue expression). If signal remains weak, test citrate at pH 6.0 as a fallback on paired sections, holding antibody dilution and detection conditions constant (standard IHC practice).
Can fixation explain inconsistent PRKCE IHC staining?
Target-specific sensitivity of PRKCE staining to fixative chemistry or fixation duration is unknown from the supplied evidence (PB9320 tissue-IHC caption: fixative not stated). The selected catalog image shows mouse intestine IHC on paraffin sections, but its caption gives no fixative, so it cannot establish fixation tolerance (PB9320 tissue-IHC caption). Record fixative, fixation duration and processing history, and compare sections prepared alike before changing antibody conditions (standard IHC practice). Run a matched control through EDTA retrieval at pH 8.0; evaluate both staining and morphology before attributing differences to fixation (datasheet PB9320 retrieval; standard IHC practice).
Which cellular PRKCE staining patterns should I accept in chromogenic IHC?
Assess cytoplasmic and membranous staining within identifiable cells, consistent with the reported tissue pattern for PRKCE (HPA: cytoplasmic and membranous expression). Perinuclear, cytoskeletal and nuclear localisation are also annotated, while stimulus-associated movement to the plasma membrane has been reported in epithelial cells (UniProt Q02156 subcellular). Because PRKCE has no transmembrane segment, a membrane-associated pattern does not establish that the protein spans the membrane (UniProt Q02156 topology). Compare compartments in matched sections and record staining intensity and cell type separately; isolated edge staining or signal outside intact cells needs control-based review (standard IHC practice).
How can epitope location affect interpretation of PRKCE IHC?
The supplied record lists 0 annotated isoforms, but that annotation alone does not establish antibody specificity in tissue (UniProt Q02156 isoforms; standard IHC practice). PRKCE contains a C2 region at residues 1–117 and a kinase domain at 408–668, with multiple modified residues that could matter if they overlap the antibody epitope (UniProt Q02156 domains and modified residues). The PB9320 tissue-IHC caption does not identify that epitope, so avoid assigning a particular staining pattern to phosphorylation or domain accessibility from this image alone (PB9320 tissue-IHC caption). Check documented immunogen information and specificity controls before interpreting differences between compartments or processing conditions (standard IHC practice).
How should I assess PRKCE by IF alongside this IHC guide?
For a separate IF/ICC comparison, multiplex PRKCE with a marker that identifies the expected cell population, such as neuronal cells when examining cerebral cortex (HPA: High in cerebral cortex neuronal cells; standard IF practice). Choose a fluorophore channel with low measured tissue autofluorescence and inspect an unstained control in every channel before calling faint signal positive (standard IF practice). PRKCE is mainly cytosolic and associated with intermediate filaments, with additional membrane and vesicle localisation, so permeabilisation should suit the intracellular side of the epitope being tested (HPA subcellular; UniProt Q02156 topology; standard IF practice). The antibody epitope is unspecified here; establish access empirically and interpret IF compartment patterns alongside matched chromogenic IHC controls (PB9320 tissue-IHC caption; standard IF/IHC practice).
What should I check when PRKCE IHC has diffuse or patchy background?
Compare stained sections with a no-primary control before changing PRKCE antibody concentration, because detection chemistry and tissue components can contribute signal (standard IHC practice). For peroxidase-based DAB detection, verify the peroxidase block and examine endogenous pigment or precipitate under the same illumination used for scoring (standard IHC practice). Check whether diffuse staining crosses cell boundaries or concentrates at folds, cut edges or damaged areas; these patterns warrant technical review before biological interpretation (standard IHC practice). Evaluate intact neuronal cells or kidney distal tubules as positive references, while remembering that PRKCE can show cytoplasmic and membranous staining (HPA: High in cerebral cortex neuronal cells and kidney distal tubules; HPA tissue expression).
How should I quantify PRKCE staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cell population before scoring, then apply identical retrieval, detection and image settings across comparison sections (standard IHC practice). For cellular staining, report the percentage of positive cells and an H-score from 0–300, calculated from percentages at intensity grades 0–3 (standard IHC practice). For spatially restricted populations, report positive-cell density per mm² of viable tissue and record cytoplasmic and membranous scores separately (standard IHC practice; HPA: cytoplasmic and membranous tissue expression). Normalise counts to eligible cells or viable area, and compare like cell populations because HPA reports markedly different PRKCE levels across tissues (standard IHC practice; HPA tissue IHC).
How can I distinguish true PRKCE signal from an IHC artefact?
A plausible positive pattern places cytoplasmic or membranous signal in intact, identifiable cells and agrees with an appropriate positive control (HPA: cytoplasmic and membranous expression; standard IHC practice). Cerebral cortex neuronal cells and kidney distal tubules are high-expression references, whereas appendix glandular cells are reported as not detected (HPA tissue IHC). Review unexpected staining in the wrong cell population or isolated at section edges, folds or necrotic areas against adjacent morphology and a no-primary control (standard IHC practice). If DAB persists without primary antibody, investigate endogenous peroxidase or detection background before assigning PRKCE expression; nuclear staining alone needs independent support for that condition (standard IHC practice; UniProt Q02156 subcellular).
Boster reagents
Best PRKCE / Protein kinase C epsilon type IHC Antibodies
Anti-PRKCE antibodies have IHC images from mouse and rat intestine and paraffin sections of human brain and stomach, plus IF data from A549 cells (PB9320 IHC captions; A01151-1 IF caption).
Real IHC data
Anti-PKC epsilon/PRKCE Antibody ®
Cat # PB9320
Real IF data
Anti-PRKCE Antibody ®
Cat # A01151-1
PB9320 is listed for IHC in human, mouse and rat, with IHC images from mouse and rat intestine and paraffin sections of human brain and stomach (PB9320 applications/reactivity; PB9320 IHC captions). A01151-1 is listed for IF/ICC in human, mouse and rat, with an IF image from A549 cells (A01151-1 applications/reactivity; A01151-1 IF caption).
Which to pick: Choose PB9320 for chromogenic tissue IHC: it is a rabbit polyclonal listed for IHC, and its captions show paraffin-section staining; the fixative is unreported (PB9320 dilution_raw; PB9320 applications; PB9320 IHC captions). Choose A01151-1 for IF/ICC: it is a rabbit polyclonal listed for those applications, with an A549 cell IF image (A01151-1 dilution_raw; A01151-1 applications; A01151-1 IF caption). For cross-species work, both list human, mouse and rat reactivity; choose according to the application and confirm performance in the intended sample (PB9320 reactivity/applications; A01151-1 reactivity/applications).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q02156 (KPCE_HUMAN, Protein kinase C epsilon type).
- Human Protein Atlas. PRKCE tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PRKCE subcellular location (ICC-IF): Mainly localized to the cytosol and intermediate filaments. In addition localized to the plasma membrane and vesicles..
- Human Protein Atlas. PRKCE antibody validation summary (3 antibodies).
- Overexpression of microRNA-205-5p exerts suppressive effects on stem cell drug resistance in gallbladder cancer by down-regulating PRKCE. Bioscience reports 2020 — PMC7533283.
- Proteomic analysis reveals key differences between squamous cell carcinomas and adenocarcinomas across multiple tissues. Nature communications 2022 — PMC9293992.
- miR-218-5p restores sensitivity to gemcitabine through PRKCE/MDR1 axis in gallbladder cancer. Cell death & disease 2017 — PMC5520703.
- PubMed PMID:1382605 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRKCE (Protein kinase C epsilon type). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9320). Validated IHC antibody: PB9320 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet PB9320); 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 PB9320); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9320); 8. Primary antibody: Rabbit anti-PRKCE, 0.5-1μg/ml (datasheet PB9320); 9. Primary incubation: Overnight at 4 °C (datasheet PB9320). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9320); Caveat: HGF can shift epithelial PRKCE to the plasma membrane (UniProt); Isoform / epitope: No listed isoforms or cleavage; no extracellular domain (UniProt); Regulation: HGF-dependent membrane redistribution (UniProt). Evidence: UniProt Q02156, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol and intermediate filaments. In…, Datasheet PB9320, PMC7533283, PMC9293992. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRKCE/immunohistochemistry-ihc-guide.