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PRKCA / Protein kinase C alpha type · IHC design guide
Design Immunohistochemistry for PRKCA
Plan chromogenic PRKCA IHC in paraffin sections with tissue controls, fixation, detection, and scoring guidance. Expect mainly cytoplasmic staining and high CNS expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRKCA 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 | Cytoplasm in most tissues; Purkinje cell membrane (HPA tissue IHC) | |
| Staining pattern | Mostly cytoplasmic; strong CNS neuronal staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9318) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Breast+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); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image PB9318) | |
| Caveat | Staining and RNA show only medium consistency (HPA tissue IHC) | |
| Regulation | Elevated in some cancer cells (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; chain 2–672; no TM segment (UniProt) |
Section 1
Recommended PRKCA IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published PRKCA staining protocols for NP tissue, rat brain, and mouse lung (PMC10249188; PMC10707860; PMC3895109).
| Sample | Paraffin-embedded mouse intestine tissues; fixative not specified (datasheet PB9318) |
| Fixation | Image fixative and duration unreported (datasheet PB9318); 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 PB9318) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9318) |
| Primary antibody | Rabbit anti-PRKCA, 0.5-1μg/ml (datasheet PB9318) |
| Primary incubation | Overnight at 4 °C (datasheet PB9318) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9318) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRKCA-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues, high expression in CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: PB9318); consider EDTA for NP tissue sections (PMC10249188: 20 min).
Section 2
What Is the Expected PRKCA Staining Pattern?
PRKCA should appear mainly in the cytoplasm, with membrane staining in some cells; it has no transmembrane segment (HPA: tissue IHC; UniProt P17252: topology). High staining is reported in adrenal glandular cells, cerebellar Purkinje cells and hippocampal neurons (HPA: tissue IHC). HPA rates the tissue IHC pattern Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Strong cytoplasmic staining in Purkinje cells, with some membrane staining. | This matches the reported cerebellar pattern (HPA: Purkinje cells, High, cytoplasm/membrane). Judge the cell and compartment together; membrane staining alone is less informative than the full pattern (HPA: tissue IHC). |
| Strong signal in adrenal glandular cells or hippocampal neurons. | Both are useful known-positive reference populations (HPA: High in adrenal glandular cells and hippocampal neurons). Compare their staining with adjacent cells rather than assigning one intensity threshold to every tissue (HPA: tissue IHC). |
| Predominantly nuclear staining where cytoplasmic staining was expected. | Check for an artefact or a context-dependent distribution before calling it specific: tissue IHC is mainly cytoplasmic (HPA: tissue IHC), while nuclear localisation is recorded for PRKCA (UniProt P17252: subcellular location). |
| Strong staining in breast adipocytes or parathyroid glandular cells. | These populations were not detected in the reported tissue IHC data (HPA: breast adipocytes; parathyroid glandular cells). Investigate cross-reactivity or endogenous chromogen activity (standard IHC practice); the HPA result is a comparator, not proof that every specimen must be negative. |
| Uniform colour across cells and extracellular areas, or no signal in a known-positive tissue. | Uniform colour suggests background from detection or washing (standard IHC practice). A blank positive reference makes a negative test section uninterpretable until the staining run is checked (standard IHC practice; HPA: High reference populations). |
💡Expected PRKCA appearanceCall a positive result when cytoplasmic, sometimes membrane-associated, staining is clear in the expected cells, especially the High Purkinje, adrenal glandular or hippocampal neuronal populations; broad colour outside cells or dominant staining of reported negative populations is suspect (HPA: tissue IHC; standard IHC practice).
| Tissue and cell choice | HPA reports High staining in Purkinje cells, hippocampal neurons and adrenal glandular cells, but Low staining in endometrial glandular cells (HPA: tissue IHC). A weak section from a Low population is a poor sole positive control. |
| Compartment and topology | HPA tissue staining is mainly cytoplasmic, and Purkinje staining includes membrane signal (HPA: tissue IHC). PRKCA lacks a transmembrane segment; membrane association should not be interpreted as evidence of an integral membrane protein (UniProt P17252: topology). |
| Strength of IHC evidence | HPA assigns Enhanced IHC status to HPA006563, HPA006564, CAB003844 and CAB016290 (HPA: antibodies). The tissue profile has medium staining–RNA consistency, so compare a questionable pattern with expected cell populations and controls (HPA: tissue IHC; standard IHC practice). |
| IF/ICC Q: What localisation is expected? | A: Mainly cytosolic signal, with supported plasma membrane signal; vesicle localisation is uncertain (HPA: subcellular ICC-IF). This is an IF/ICC interpretation cue, not an IHC-P staining condition. |
| Situation | Likely cause | Next action |
|---|---|---|
| A known-positive section has no visible PRKCA signal. | The run may have failed, or the chosen area may lack the reported positive cells (standard IHC practice; HPA: tissue IHC). | Check that Purkinje cells, hippocampal neurons or adrenal glandular cells are present; review the run's controls, antibody application, retrieval and detection steps (HPA: High populations; standard IHC practice). |
| A test section is weak while the positive reference stains clearly. | The sampled population may normally stain at a lower level; HPA reports Low staining in endometrial glandular cells (HPA: tissue IHC). | Score identifiable cells against the positive reference and the relevant HPA population; avoid increasing detection solely to force a High-like result in a Low population (HPA: tissue IHC; standard IHC practice). |
| The slide has widespread diffuse brown background. | Nonspecific detection, endogenous activity or inadequate washing can produce diffuse colour (standard chromogenic IHC practice). | Review reagent and no-primary controls, blocking and wash steps; assess whether signal remains confined to cells with an expected pattern (standard IHC practice; HPA: tissue IHC). |
| Signal is predominantly nuclear. | The tissue profile is mainly cytoplasmic, although UniProt also records nuclear localisation (HPA: tissue IHC; UniProt P17252: subcellular location). | Compare the same run's positive reference and controls before treating nuclear signal as specific; record the specimen context if the pattern persists (standard IHC practice). |
| Reported negative cell populations stain strongly. | Cross-reactivity or endogenous detection activity is possible (standard IHC practice); HPA reports no detection in breast adipocytes and parathyroid glandular cells (HPA: tissue IHC). | Confirm cell identity and review no-primary and detection controls. Treat the HPA negatives as reference observations rather than universal absence claims (HPA: tissue IHC; standard IHC practice). |
| Only slide edges or isolated damaged areas stain intensely. | Uneven reagent coverage or local section damage can distort a chromogenic pattern (standard IHC practice). | Inspect intact central tissue and control sections, then repeat with even reagent coverage if the pattern cannot be scored reliably (standard IHC practice). |
Experiment evidence
IHC & IF images showing PRKCA staining
PB9318 · IHCIHC analysis of PKC alpha using anti-PKC alpha antibody (PB9318). PKC alpha was detected in paraffin-embedded section of mouse intestine tis
PB9318 · IHCIHC analysis of PKC alpha using anti-PKC alpha antibody (PB9318). PKC alpha was detected in paraffin-embedded section of rat intestine tissu
PA1065 · IHCIHC analysis of PKC alpha using anti-PKC alpha antibody (PA1065). PKC alpha was detected in a paraffin-embedded section of Human Mammary Can
M00743 · IHCImmunohistochemical analysis of paraffin-embedded Human pituitary tumor, using the Antibody at 1:50 dilution.
M00743 · IHCImmunohistochemical analysis of paraffin-embedded Human thymoma, using the Antibody at 1:300 dilution.
M00743 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
M00743-6 · IHCMouse liver was stained with anti-PKC α rabbit antibody
M00743-6 · IHCRat liver was stained with anti-PKC α rabbit antibody
A00743T495 · IHCImmunohistochemistry (IHC) analyzes of PKC-pan (T495) pAb in paraffin-embedded human breast carcinoma tissue at 1:100.
A00743T497 · IHCImmunohistochemistry (IHC) analyzes of p-PKC-pan (αT497) pAb in paraffin-embedded human breast carcinoma tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000154229-PRKCA/tissue/cerebral+cortex#imid_2007408
HPA · HippocampusHuman Protein Atlas IHC, Hippocampus. Source: https://www.proteinatlas.org/ENSG00000154229-PRKCA/tissue/hippocampal+formation#imid_2007425
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000154229-PRKCA/tissue/colon#imid_2007350
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000154229-PRKCA/tissue/liver#imid_2007343
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006563; green = PRKCA, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA006563; green = PRKCA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRKCA IHC & IF
🧪Run cerebellum first: Purkinje cells should show strong cytoplasmic or membrane staining (HPA: High in Purkinje cells). Run parathyroid gland as the negative tissue (HPA: Not detected in glandular cells); on the cerebellum slide, cells without specific staining should show only background chromogen, though HPA does not establish them as PRKCA-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Breast (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRKCA in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a concentration-matched rabbit isotype control (PB9318 caption: rabbit primary), and PRKCA knockout tissue as a biological negative. With SABC/DAB detection, check endogenous peroxidase and biotin background in cerebellum (PB9318 caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the PB9318 paraffin-section caption does not state the fixative. Heat retrieval in citrate buffer at pH 6 for 20 minutes was used for mouse intestine; whether cerebellum staining depends on that retrieval is unreported (PB9318 caption). The supplied evidence does not establish that frozen sections or IF are easier; endogenous biotin can produce background with SABC detection in cerebellum (PB9318 caption: SABC; standard IHC practice).
HPA tissue IHC evidence for PRKCA
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skin | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PRKCA IHC Tips
Troubleshoot PRKCA staining in paraffin sections by checking retrieval, compartment, cell type, detection background, and scoring against the available tissue evidence.
How should I adjust retrieval when PRKCA staining is weak?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet PB9318). The selected paraffin-section example used this treatment before staining mouse intestine with 1 µg/mL antibody overnight at 4°C (datasheet PB9318). If staining is weak, compare a modestly shorter and longer heating period on serial sections while keeping buffer and detection conditions constant (standard IHC practice). Check tissue integrity and background alongside signal, because excessive heating can damage morphology or increase nonspecific staining (standard IHC practice). Document the actual heating conditions and compare expected cytoplasmic staining in the same anatomical compartments (HPA tissue IHC profile).
Could fixation explain weak or uneven PRKCA staining?
PRKCA-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state its fixative (datasheet PB9318). Record the fixative, fixation duration, section thickness, and time before processing for each specimen, then compare sections handled under matched conditions (standard IHC practice). If staining varies across a batch, review fixation and retrieval together before increasing antibody concentration, since both affect antigen accessibility in paraffin-section IHC (standard IHC practice). Use the documented citrate pH 6, 20-minute retrieval as a starting condition (datasheet PB9318). Tissue staining patterns alone cannot establish how fixation affects this antibody (HPA tissue IHC profile).
Which compartments should show convincing PRKCA staining?
Expect predominantly cytoplasmic staining in tissue sections, with cytoplasm and membrane staining specifically reported in cerebellar Purkinje cells (HPA tissue IHC profile). PRKCA can also occupy the cell membrane, mitochondrial membrane, and nucleus (UniProt P17252 subcellular annotation). Because PRKCA has no transmembrane segment, a membrane-associated pattern should be assessed as localisation of a peripheral kinase rather than evidence of a membrane-spanning protein (UniProt P17252 topology). Score cytoplasmic, membranous, and nuclear signal separately when compartment shifts are the question (standard IHC practice). Interpret isolated nuclear staining in its biological context: nuclear translocation is described during macrophage differentiation, but it is not a universal baseline pattern (UniProt P17252 function).
Could epitope selection change the apparent PRKCA pattern?
Check the antibody's stated immunogen and epitope before comparing it with another PRKCA reagent; the supplied caption does not identify either (datasheet PB9318). The record lists 0 annotated isoforms, so these data do not support an isoform-specific explanation for discordant staining (UniProt P17252 isoform annotation). PRKCA has a C2 domain at residues 158–275 and a kinase domain at 339–597, with multiple annotated modified residues (UniProt P17252 domains and modified residues). An epitope near a modified site could merit investigation, but the payload does not show that phosphorylation alters this antibody's IHC binding (UniProt P17252 modified residues; datasheet PB9318). Compare antibody results by compartment and cell type under matched processing conditions (standard IHC practice).
How can IF help resolve ambiguous PRKCA localisation?
Use IF as a separate localisation check and multiplex PRKCA with a marker identifying the cell population under study (standard IF practice). Cytosol is the supported main location; plasma membrane is additionally supported, whereas vesicle localisation is uncertain (HPA subcellular profile). Choose fluorophores after inspecting tissue autofluorescence in unstained controls, and place the weaker signal in a spectrally cleaner channel when possible (standard IF practice). Match permeabilisation to the proposed epitope: access to cytosolic PRKCA generally requires membrane permeabilisation, while the antibody's epitope side is not specified here (UniProt P17252 topology; datasheet PB9318). Do not treat the paraffin IHC caption as evidence of IF fixation or staining performance (datasheet PB9318).
What should I check when DAB obscures PRKCA staining?
First inspect a section processed without primary antibody to locate detection-system background (standard IHC practice). The selected example used 10% goat serum block, biotinylated goat anti-rabbit secondary antibody, a streptavidin-biotin complex, and DAB (datasheet PB9318). Include an endogenous peroxidase block before DAB development and assess whether endogenous biotin contributes signal with this detection scheme (standard IHC practice). Titrate antibody and shorten chromogen development if diffuse brown staining overwhelms cellular detail, changing one condition at a time (standard IHC practice). Compare results with the expected broadly cytoplasmic tissue pattern and appropriate cellular controls (HPA tissue IHC profile; standard IHC practice).
How should I quantify PRKCA across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the anatomical compartment and cell population before scoring, since PRKCA expression varies among tissues and cell types (HPA tissue IHC profile). For cellular staining, record the percentage of positive cells and an H-score based on intensity categories; score cytoplasmic, membrane, and nuclear signal separately when relevant (standard IHC practice; UniProt P17252 subcellular annotation). If the question concerns infiltrating positive cells, report positive-cell density per mm² of viable tissue (standard IHC practice). Normalise comparisons to the same cell population or viable tissue area, using matched section thickness, imaging, and detection settings (standard IHC practice). Apply one prespecified positivity threshold to all compared sections (standard IHC practice).
When is PRKCA staining likely to be artefactual?
A convincing result places signal in plausible cells and compartments: cytoplasm is common across tissues, while cerebellar Purkinje cells can show cytoplasmic and membranous staining (HPA tissue IHC profile). Treat staining confined to cut edges or necrotic areas cautiously, especially if comparable signal appears in a no-primary control (standard IHC practice). Diffuse DAB deposits may reflect endogenous enzyme activity or detection background; verify the peroxidase block and inspect controls before assigning them to PRKCA (standard IHC practice). An exclusively extracellular pattern warrants review because PRKCA is annotated in intracellular and membrane-associated compartments and has no transmembrane segment (UniProt P17252 subcellular annotation and topology). Reconcile unexpected cells with tissue context rather than declaring every brown cell positive (HPA tissue IHC profile; standard IHC practice).
Boster reagents
Best PRKCA / Protein kinase C alpha type IHC Antibodies
IHC images cover human and mouse tissues; the catalog lists human, mouse, and rat reactivity, and M00743 has IF images (catalog image captions; catalog reactivity and IF images).
Real IHC data
Anti-PKC alpha/PRKCA Antibody ®
Cat # PB9318
Real IHC data
Anti-PKC alpha/PRKCA Antibody ®
Cat # PA1065
Real IHC data
Anti-PKC alpha PRKCA Rabbit Monoclonal Antibody
Cat # M00743
Real IHC data
Anti-PKC α Rabbit Monoclonal Antibody
Cat # M00743-6
Real IHC data
Anti-PKC-pan (T495) PRKCA Antibody
Cat # A00743T495
Real IHC data
Anti-PKC-pan ( Alpha T497) PRKCA Antibody
Cat # A00743T497
PB9318 shows IHC in mouse intestine, PA1065 in human mammary cancer, and M00743 in human pituitary tumor (respective IHC image captions). M00743-6 shows mouse liver, A00743T495 shows human breast carcinoma, and A00743T497 shows human breast carcinoma with a phospho-PKC-pan antibody (respective IHC image captions).
Which to pick: For paraffin-section IHC, choose PB9318 for its mouse intestine example or PA1065 for its human mammary cancer example; neither caption reports the fixative (respective IHC image captions). For IF/ICC, choose M00743: its application list includes both, it is a rabbit monoclonal, and it has IF images (M00743 catalog applications, clone, and IF image captions). For work across human, mouse, and rat, PB9318 has IHC captions for all three species; M00743-6 also has human, mouse, and rat reactivity and is listed as a recombinant rabbit monoclonal (PB9318 IHC image captions; M00743-6 catalog reactivity and description).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P17252 (KPCA_HUMAN, Protein kinase C alpha type).
- Human Protein Atlas. PRKCA tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PRKCA subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the plasma membrane and vesicles..
- Human Protein Atlas. PRKCA antibody validation summary (4 antibodies).
- tsRNA-04002 alleviates intervertebral disk degeneration by targeting PRKCA to inhibit apoptosis of nucleus pulposus cells. Journal of orthopaedic surgery and research 2023 — PMC10249188.
- Clinical significance of the correlation between PLCE 1 and PRKCA in esophageal inflammation and esophageal carcinoma. Oncotarget 2017 — PMC5464868.
- Network Analysis and Experimental Verification of the Mechanisms of Hydroxysafflor Yellow A in Ischemic Stroke Following Atherosclerosis. Molecules (Basel, Switzerland) 2023 — PMC10707860.
- Protein kinase Cα suppresses Kras-mediated lung tumor formation through activation of a p38 MAPK-TGFβ signaling axis. Oncogene 2014 — PMC3895109.
- PubMed PMID:2336401 — UniProt-cited evidence.
- PubMed PMID:19054851 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRKCA (Protein kinase C alpha type). Expected localisation Cytoplasm in most tissues; Purkinje cell membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9318). Validated IHC antibody: PB9318 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Breast. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse intestine tissues; fixative not specified (datasheet PB9318); 2. Fixation: Image fixative and duration unreported (datasheet PB9318); 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 PB9318); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9318); 8. Primary antibody: Rabbit anti-PRKCA, 0.5-1μg/ml (datasheet PB9318); 9. Primary incubation: Overnight at 4 °C (datasheet PB9318). Watch for: Fixation: Keep fixation consistent across sections (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 PB9318); Caveat: Staining and RNA show only medium consistency (HPA tissue IHC); Isoform / epitope: 0 annotated isoforms; chain 2–672; no TM segment (UniProt); Regulation: Elevated in some cancer cells (UniProt). Evidence: UniProt P17252, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the plasma…, Datasheet PB9318, PMC10249188, PMC10707860. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRKCA/immunohistochemistry-ihc-guide.