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PRKCI / Protein kinase C iota type · IHC design guide
Design Immunohistochemistry for PRKCI
Plan PRKCI chromogenic IHC around the cytoplasmic staining reported in most tissues (HPA tissue IHC). Use the catalog antibody's IHC-P dilution of 0.5–1 μg/ml (datasheet) and compare staining with a matched control section (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRKCI 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 in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Fallopian tube+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 low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | High expression in non-small cell lung cancer (UniProt) | |
| Isoform / epitope | No listed isoforms; mature chain spans aa 2–596 (UniProt) |
Section 1
Recommended PRKCI IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published PRKCI protocol using mouse paraffin sections (PMC5538434).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet PB9321); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-PRKCI, 0.5-1μg/ml (datasheet PB9321) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRKCI-positive staining in glandular cells of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page antigen retrieval); compare the published citrate retrieval if needed (PMC5538434).
Section 2
What Is the Expected PRKCI Staining Pattern?
PRKCI should show mainly cytoplasmic staining in many tissues, including glandular and epithelial cells (HPA tissue IHC: cytoplasmic expression in most tissues). Membrane, endosomal and nuclear localization are also reported, and PRKCI has no transmembrane segment (UniProt P41743). Interpret staining cautiously: the tissue IHC assessment is Approved but shows low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in fallopian tube or stomach glandular cells, or tonsil squamous epithelial cells. | This fits the reported High staining in those cell types and the predominant tissue pattern (HPA tissue IHC). Judge the signal within the identified cells; staining of every structure in the section is less convincing (general IHC practice). |
| Cytoplasmic staining with a localized membrane associated, punctate, or nuclear component. | These compartments are plausible because PRKCI is reported in cytoplasm, membrane, endosomes and nucleus (UniProt P41743). A nuclear component alone is not proof of specificity; assess its cell distribution and controls (general IHC practice). |
| Staining confined to extracellular material or other compartments without a credible cellular pattern. | An exclusively extracellular pattern does not fit the reported cellular localization (UniProt P41743; HPA tissue IHC). Consider precipitated chromogen, nonspecific binding or tissue debris, then inspect a detection control (general IHC practice). |
| Strong signal in cells reported as undetected, such as adipocytes or bone marrow hematopoietic cells. | This conflicts with those cell specific observations (HPA tissue IHC: Not detected). Cross reactivity or endogenous detection activity is possible; compare morphology and matched controls before assigning PRKCI positivity (general IHC practice). |
| No signal in fallopian tube or stomach glandular cells while section morphology and controls remain interpretable. | These are reported High staining examples (HPA tissue IHC). Check the assay and antibody performance before treating a negative result as biological absence; HPA also reports low staining to RNA consistency (HPA tissue IHC; general IHC practice). |
💡Expected PRKCI appearanceA convincing positive is chiefly cytoplasmic, with High staining possible in fallopian tube or stomach glandular cells and tonsil squamous epithelium; uniform extracellular color or strong signal in reported undetected cell types calls for control review (HPA tissue IHC; UniProt P41743; general IHC practice).
| Tissue and cell selection (HPA tissue IHC) | Fallopian tube and stomach glandular cells and tonsil squamous epithelial cells are High; bronchial respiratory epithelium and cerebral cortex neurons are Medium (HPA tissue IHC). Compare the named cell type, not the whole tissue average (general IHC practice). |
| Intracellular distribution (UniProt P41743) | PRKCI is reported in cytoplasm, membrane, endosomes and nucleus (UniProt P41743). SQSTM1 interaction is associated with endosomal transport, and SRC phosphorylation with KPNB1 mediated nuclear transport (UniProt P41743). These reports allow localized departures from an otherwise cytoplasmic pattern. |
| Topology and processing (UniProt P41743) | PRKCI has no transmembrane segment or signal peptide; the annotated protein chain spans residues 2–596 (UniProt P41743). An exclusively extracellular deposit is therefore a poor localization match. This annotation does not predict antibody epitope accessibility in paraffin sections. |
| Antibody evidence (HPA antibodies; HPA tissue IHC) | One listed antibody, HPA026574, has Approved IHC status; HPA038635 has no listed IHC status (HPA antibodies). The tissue profile is also Approved but has low staining to RNA consistency, so individual unexpected cells warrant control based review (HPA tissue IHC). |
| Assay chemistry (general IHC practice) | In chromogenic IHC, endogenous enzyme activity, nonspecific secondary binding or chromogen deposits can add color unrelated to primary antibody binding (general IHC practice). A detection control helps separate these sources from the cellular pattern; no PRKCI specific fixation sensitivity is supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in a reported High cell population (HPA tissue IHC). | The assay may have failed, or staining may fall below detection; the HPA pattern alone cannot identify which (HPA tissue IHC; general IHC practice). | Check tissue preservation, primary antibody identity, detection reagents and a known positive control; review retrieval against the IHC validated antibody's instructions (general IHC practice). |
| Uniform weak color obscures cell boundaries. | Background can arise from nonspecific binding, residual detection activity or excess chromogen development (general IHC practice). | Compare a matched detection control; review blocking, washes, antibody concentration and development time using the IHC assay instructions (general IHC practice). |
| Dark deposits appear outside recognizable cells. | Precipitated chromogen or tissue debris may mimic signal; extracellular deposition does not match annotated PRKCI localization (general IHC practice; UniProt P41743). | Inspect the section at higher magnification and compare the same area with a detection control and counterstained morphology (general IHC practice). |
| Strong staining appears in adipocytes or bone marrow hematopoietic cells. | Both are reported Not detected; cross reactivity or endogenous detection activity is possible, though an HPA result is not an absolute exclusion (HPA tissue IHC; general IHC practice). | Confirm cell identity and compare matched negative and positive controls before scoring those cells as PRKCI positive (general IHC practice). |
| A sample shows nuclear staining alongside cytoplasmic staining. | Nuclear localization is reported after SRC associated transport through KPNB1, while cytoplasm is also reported (UniProt P41743). The staining pattern alone does not establish that pathway's activity. | Record the compartments separately and assess the nuclear signal against morphology and controls; avoid scoring it as an artifact solely because it is nuclear (UniProt P41743; general IHC practice). |
| IF/ICC question: should fluorescence match paraffin IHC exactly? | The assays sample different preparations; HPA reports mainly cytosolic ICC/IF signal, with additional microtubule, cytokinetic bridge and primary cilium localization (HPA subcellular). | Use the separate IF/ICC guide for that assay. Compare each result with its own localization evidence and controls rather than requiring identical compartment patterns (HPA subcellular; HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing PRKCI staining
PB9321 · IHCAnti-PKC iota vendor antibody, PB9321,IHC(P) IHC(P): Mouse Brain Tissue
PB9321 · IHCAnti-PKC iota vendor antibody, PB9321,IHC(P) IHC(P): Rat Brain Tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163558-PRKCI/tissue/cerebral+cortex#imid_6542623
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163558-PRKCI/tissue/colon#imid_6543348
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163558-PRKCI/tissue/liver#imid_6543341
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163558-PRKCI/tissue/kidney#imid_6543342
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026574; green = PRKCI, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026574; green = PRKCI, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRKCI IHC & IF
🧪Run stomach first: its glandular cells should stain (HPA: High in stomach glandular cells). Use adipose tissue as a negative comparator, focusing on adipocytes (HPA: Not detected in adipocytes); on the stomach slide, score neighboring cells without signal as internal negatives only after confirming their identity, since the supplied HPA row establishes staining only for glandular cells.
Positive control tissue: Fallopian tube (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRKCI in A-431, U-251MG, U2OS, MCF-7, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), KOLF2.1J, NIH 3T3, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a nonspecific IgG control matched to the primary antibody’s host species and, for a monoclonal antibody, its isotype. Use PRKCI-knockout material or an immunizing-peptide block, if available, as a biological specificity control; for stomach IHC, quench endogenous peroxidase and inspect gland lumens for nonspecific chromogen before scoring (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence; the selected PB9321 mouse-brain IHC(P) caption also leaves its fixative unreported (selected-SKU caption: fixative not stated). Retrieval dependency is unreported, so compare retrieval conditions using the positive and negative tissues rather than assuming one is required (HPA: High in stomach glandular cells; Not detected in adipocytes). The evidence does not establish that frozen sections or IF are easier; for IF interpretation, cytosolic signal is expected (HPA subcellular: Cytosol supported), and stomach glandular signal should be distinguished from nonspecific luminal chromogen in IHC (standard IHC practice).
HPA tissue IHC evidence for PRKCI
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Stomach | Glandular cells | High | Protein (IHC) | HPA → |
| Tonsil | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PRKCI IHC Tips
Troubleshoot PRKCI staining in paraffin sections by checking retrieval, cell compartment, antibody specificity and matched controls before scoring chromogenic signal.
How should I retrieve PRKCI in paraffin sections when staining is weak?
Start with Tris-EDTA, pH 9.0, heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Keep heating, cooling and section thickness consistent across control and study slides so changes in staining can be compared (standard IHC practice). If staining remains weak, test a citrate buffer at pH 6.0 on adjacent sections as a fallback, with the same antibody and detection conditions (standard IHC practice). Compare signal and tissue morphology in a positive control, such as HPA-reported fallopian tube glandular cells (HPA: High), rather than judging intensity alone. An altered retrieval result does not establish which PRKCI epitope the catalog antibody recognizes (record: epitope unspecified).
Could fixation explain weak or uneven PRKCI staining?
Target-specific PRKCI sensitivity to fixation is unknown from the supplied evidence; the selected PB9321 IHC(P) mouse brain caption does not state a fixative (caption: PB9321). Record the fixative, fixation duration and processing history for each block before comparing staining across samples (standard IHC practice). When possible, stain adjacent sections from differently processed material in one run, holding retrieval, antibody incubation and detection constant (standard IHC practice). Review nuclear detail and section integrity alongside PRKCI signal, because damaged morphology makes cellular assignment unreliable (standard IHC practice). Do not use HPA staining patterns or PRKCI topology and modifications to infer a fixation effect (HPA: tissue profile; UniProt P41743: topology and modified residues).
Which staining compartments are plausible for PRKCI in IHC?
Expect cytoplasmic staining as the broad tissue pattern, and assess any membrane or nuclear signal separately (HPA: cytoplasmic expression in most tissues; UniProt P41743: cytoplasm, membrane, nucleus). PRKCI has no transmembrane segment, so membranous staining should be interpreted as localization rather than evidence of a membrane-spanning protein (UniProt P41743: topology). Endosomal localization is reported with SQSTM1 interaction, while SRC phosphorylation and KPNB1 interaction are associated with nuclear transport (UniProt P41743: subcellular location). Score each compartment within identifiable cells and compare it with the corresponding negative control (standard IHC practice). Diffuse color outside intact cells or concentrated at section edges warrants a staining-artifact check (standard IHC practice).
How can I assess whether staining reflects PRKCI rather than another epitope?
The supplied record lists 0 isoforms but does not identify the catalog antibody's immunogen or epitope (UniProt P41743: isoforms; record: epitope unspecified). PRKCI contains a PB1 region at residues 25–108, a kinase domain at 254–522 and an AGC C-terminal region at 523–594 (UniProt P41743: domains). Its recorded modifications include phosphorylation near the N terminus, but their effect on this antibody's staining is unknown without epitope information (UniProt P41743: modified residues; record: epitope unspecified). Check the antibody's documented immunogen before making claims about domain or modification sensitivity (standard antibody-validation practice). Compare staining with an independently validated PRKCI antibody or a suitable loss-of-target control where available (standard antibody-validation practice).
How should I follow up a chromogenic PRKCI pattern with IF?
Use IF/ICC as a separate validation experiment; the supplied PB9321 tissue image documents IHC(P) in mouse brain, not IF performance (caption: PB9321). Multiplex PRKCI with a marker of the cell population being assessed, and check each channel separately for bleed-through (standard IF practice). Choose fluorophores and detection channels after examining unstained tissue autofluorescence, particularly if the chromogenic section shows pigment or strong background (standard IF practice). Because PRKCI lacks a transmembrane segment and has cytoplasmic and nuclear locations, optimize permeabilisation for intracellular access without assuming the antibody epitope is known (UniProt P41743: topology and location; record: epitope unspecified). Compare the resulting cellular pattern with matched IHC sections and IF controls (standard IF practice).
What should I check when PRKCI staining is widespread or granular?
First inspect a no-primary control to separate detection-system color from antibody-dependent staining (standard IHC practice). Quench endogenous peroxidase before chromogenic detection and compare the no-primary slide, especially where pigment or blood products complicate interpretation (standard IHC practice). Titrate the catalog antibody and review blocking, washes and development time on adjacent sections while keeping retrieval fixed (standard IHC practice). Cytoplasmic signal across many tissues can be plausible for PRKCI, but that pattern alone does not establish specificity (HPA: cytoplasmic expression in most tissues; HPA: low staining–RNA consistency). Reassess granular deposits against tissue architecture and an independent control before assigning them to PRKCI-positive endosomes (UniProt P41743: endosome; standard IHC practice).
How should I score PRKCI IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; PRKCI can be cytoplasmic, membranous or nuclear (UniProt P41743: subcellular location). For cellular staining, report the percentage of positive cells and an H-score from intensity categories 0–3, using the same threshold across slides (standard IHC scoring practice). If counting positive cells per area, report density per mm² of viable, evaluable tissue rather than the whole scanned section (standard image-analysis practice). Normalize comparisons to the number of eligible cells or the measured viable area within the same annotated compartment (standard IHC scoring practice). Keep acquisition, counterstain and chromogen-development settings consistent, and document exclusions for folds, edges and necrosis (standard IHC practice).
When is a PRKCI-positive IHC result convincing?
A convincing result localizes within intact, identifiable cells and recurs in a matched control without corresponding no-primary signal (standard IHC practice). Cytoplasmic staining agrees with the broad HPA tissue profile, while nuclear or membrane staining is biologically plausible but requires compartment-specific review (HPA: cytoplasmic expression in most tissues; UniProt P41743: location). HPA reports High staining in fallopian tube glandular cells and no detection in adipocytes, yet notes low consistency between antibody staining and RNA expression (HPA: tissue IHC). Exclude section-edge enhancement, necrotic areas, pigment and endogenous enzyme color before assigning a positive score (standard IHC practice). Confirm unexpected cell types or compartments with independent antibody or loss-of-target evidence where feasible (standard antibody-validation practice).
Boster reagents
Best PRKCI / Protein kinase C iota type IHC Antibodies
PB9321 has IHC images from mouse and rat brain and human lung cancer tissue (catalog IHC image captions). Its listed reactivity is human, mouse, and rat (catalog applications/reactivity); no IF data are supplied (catalog).
Real IHC data
Anti-PKC iota/PRKCI Antibody ®
Cat # PB9321
PB9321 is the only card and lists IHC for human, mouse, and rat (catalog applications/reactivity). Its IHC(P) captions show mouse brain, rat brain, and human lung cancer tissue (catalog IHC image captions).
Which to pick: Choose PB9321 for paraffin-section tissue IHC: its own IHC(P) captions show mouse brain, rat brain, and human lung cancer tissue (catalog IHC image captions); the fixative is unreported (catalog). It is a rabbit antibody listed as reactive with human, mouse, and rat, making it the available cross-species choice (catalog host/reactivity). No listed antibody supports an IF/ICC recommendation here because PB9321 has no IF application or image in the payload (catalog applications/IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P41743 (KPCI_HUMAN, Protein kinase C iota type).
- Human Protein Atlas. PRKCI tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PRKCI subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the microtubules, cytokinetic bridge and primary cilium..
- Human Protein Atlas. PRKCI antibody validation summary (2 antibodies).
- Integrated genomic analysis of clear cell ovarian cancers identified PRKCI as a potential therapeutic target. Oncotarget 2017 — PMC5722499.
- Clinical significance of atypical protein kinase C (PKCι and PKCζ) and its relationship with yes-associated protein in lung adenocarcinoma. BMC cancer 2019 — PMC6693135.
- Downregulation of PRKCI inhibits osteosarcoma cell growth by inactivating the Akt/mTOR signaling pathway. Frontiers in oncology 2024 — PMC11249533.
- PRKCI promotes immune suppression in ovarian cancer. Genes & development 2017 — PMC5538434.
- PubMed PMID:8226978 — UniProt-cited evidence.
- PubMed PMID:7607695 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRKCI (Protein kinase C iota type). Expected localisation Cytoplasmic in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: PB9321 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Glandular 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 PB9321); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-PRKCI, 0.5-1μg/ml (datasheet PB9321); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No listed isoforms; mature chain spans aa 2–596 (UniProt); Regulation: High expression in non-small cell lung cancer (UniProt). Evidence: UniProt P41743, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet PB9321, PMC5538434. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRKCI/immunohistochemistry-ihc-guide.