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PIK3CG / Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform · IHC design guide
Design Immunohistochemistry for PIK3CG
Plan paraffin-section PIK3CG IHC with the catalog antibody at 2–5 μg/ml (datasheet A01517-2). Interpret cytoplasmic and membranous staining across tissue types while accounting for presumed off-target binding (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PIK3CG 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 | Cytoplasmic/membranous staining across several tissue types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01517-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may confound staining (HPA tissue IHC) | |
| Regulation | Stimulus-dependent expression is unreported (UniProt) | |
| Isoform / epitope | 0 listed isoforms; one full-length intracellular chain (UniProt) |
Section 1
Recommended PIK3CG IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published PIK3CG IHC protocol for hepatocellular carcinoma tissue (PMC8805872).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01517-2) |
| Fixation | Image fixative and duration unreported (datasheet A01517-2); 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 A01517-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01517-2) |
| Primary antibody | Rabbit anti-PIK3CG, 2-5μg/ml (datasheet A01517-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01517-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01517-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PIK3CG-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and membranous expression in several different tissue types, including lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A01517-2); the published protocol does not specify retrieval (PMC8805872).
Section 2
What Is the Expected PIK3CG Staining Pattern?
PIK3CG staining should be cytoplasmic and membranous in several tissue types, including lymphoid tissues (HPA: tissue IHC profile). Bone marrow hematopoietic cells and several glandular cell populations show medium staining (HPA: tissue IHC). A cell membrane signal is plausible for this cytoplasmic, membrane associated protein without a transmembrane segment (UniProt P48736: localization and topology). Interpret the pattern with care: HPA rates tissue IHC reliability as Enhanced but reports presumed off target binding (HPA: reliability description).
| Cytoplasmic staining, with membrane accentuation, in bone marrow hematopoietic cells. | This fits a documented medium staining cell population and the reported compartments (HPA: bone marrow IHC; HPA: tissue IHC profile). Assess the hematopoietic cells themselves; staining elsewhere in the section does not establish that they are positive (general IHC practice). |
| Predominantly nuclear staining with little cytoplasmic or membrane signal. | This does not fit the reported localization (UniProt P48736: cytoplasm and cell membrane; HPA: tissue IHC profile). Treat an isolated nuclear pattern as suspect and compare its distribution with controls before scoring it as PIK3CG (general IHC practice). |
| Prominent staining in adipocytes or cardiomyocytes. | HPA reports PIK3CG as not detected in these cells (HPA: adipose tissue and heart muscle IHC). Review cell identity, nearby infiltrating cells and control staining; off target antibody binding or endogenous detection activity could explain the discrepancy (HPA: reliability description; general IHC practice). |
| Chromogen haze spreads across cells, stroma and blank areas. | A field wide deposit lacks the cell and compartment pattern described for PIK3CG (HPA: tissue IHC profile). Consider background from the detection workflow, including insufficient blocking or washing, and inspect a no primary control (general IHC practice). |
| No staining in bone marrow hematopoietic cells. | This conflicts with their reported medium staining (HPA: bone marrow IHC). First confirm the cells are present and the detection controls worked; then review the IHC run before calling another specimen negative (general IHC practice). |
💡Expected PIK3CG appearanceA credible positive result is cytoplasmic staining with membrane accentuation in identifiable hematopoietic or documented glandular cells, with medium staining reported for those cells; isolated nuclear staining or equally prominent staining of HPA negative cell populations is suspect (HPA: tissue IHC; UniProt P48736: localization).
| Compartment and topology | PIK3CG is cytoplasmic and associated with the cell membrane, with no transmembrane segment (UniProt P48736: localization and topology). Read membrane accentuation as localization, not evidence that the protein spans the membrane; evaluate it together with cytoplasmic staining (HPA: tissue IHC profile). |
| Cell context | HPA reports medium staining in bone marrow hematopoietic cells and in glandular cells from several organs (HPA: tissue IHC). It reports no detection in adipocytes and cardiomyocytes (HPA: tissue IHC). Score identified cells rather than assigning one result to an entire mixed tissue section (general IHC practice). |
| Evidence and antibody validation | Tissue IHC reliability is Enhanced, while the record notes medium agreement with RNA data and presumed off target binding that was disregarded (HPA: reliability description). Antibody HPA069976 is rated Enhanced for IHC and Supported for ICC (HPA: antibody validation); neither rating resolves every unexpected cell specific result. |
| Heart interpretation | UniProt lists heart in tissue specificity and describes cardiac functions (UniProt P48736: tissue specificity and function), whereas HPA reports no detection in cardiomyocytes (HPA: heart muscle IHC). Record the cell type and assay when comparing these findings; do not convert a tissue level annotation into an expected cardiomyocyte IHC positive. |
| IF/ICC Q&A: Where should signal appear? | Mainly at the plasma membrane, with additional cytosolic signal in the HPA ICC-IF record (HPA: subcellular localization). This supports a compartment check for IF/ICC; its Supported ICC rating and cell images do not set an IHC intensity threshold (HPA: antibody validation and subcellular record). |
| Situation | Likely cause | Next action |
|---|---|---|
| Bone marrow hematopoietic cells lack signal. | A run or detection failure is possible when a documented medium staining population appears blank (HPA: bone marrow IHC; general IHC practice). | Confirm cell identity, check positive and no primary controls, then review the catalog antibody's IHC-P procedure and the run's retrieval and detection records (general IHC practice). Target specific fixation sensitivity is unreported in the supplied sources. |
| Nuclei dominate the staining. | The compartment conflicts with cytoplasmic and membrane localization (UniProt P48736: localization; HPA: tissue IHC profile). | Compare nuclear staining with the no primary control and review counterstain and detection background; score PIK3CG only where the cell pattern is credible (general IHC practice). |
| Adipocytes or cardiomyocytes appear broadly positive. | These cells are reported as not detected; off target binding or endogenous detection activity is possible (HPA: adipose tissue and heart muscle IHC; HPA: reliability description; general IHC practice). | Verify cell identity and inspect neighboring hematopoietic cells. Check no primary and detection controls before accepting the unexpected signal (general IHC practice). |
| The whole section has diffuse brown haze. | Nonspecific detection or inadequate washing may obscure the reported cell pattern (HPA: tissue IHC profile; general IHC practice). | Inspect no primary and detection controls, blocking, washes and chromogen development; repeat scoring only after the background is resolved (general IHC practice). |
| A lymphoid region looks uneven or faint. | HPA describes expression in lymphoid tissues but does not assign every cell in those regions an intensity in this payload (HPA: tissue IHC profile). | Identify and score individual cell populations, then compare with a documented positive such as bone marrow hematopoietic cells from the same validated workflow (HPA: bone marrow IHC; general IHC practice). |
| IHC and IF/ICC seem to show different membrane emphasis. | HPA tissue IHC reports both cytoplasmic and membranous staining, while its ICC-IF summary says mainly plasma membrane with additional cytosol (HPA: tissue IHC profile and subcellular record). | Compare compartments within each assay and its controls; do not require identical visual intensity across chromogenic sections and fluorescent cells (general IHC/IF practice). |
Experiment evidence
IHC & IF images showing PIK3CG staining
A01517-2 · IHCIHC analysis of PI3K-gamma/PIK3CG using anti PI3K-gamma/PIK3CG antibody (A01517-2). PI3K-gamma/PIK3CG was detected in paraffin-embedded sect
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000105851-PIK3CG/tissue/cerebral+cortex#imid_20552843
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000105851-PIK3CG/tissue/colon#imid_20552753
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000105851-PIK3CG/tissue/liver#imid_20552770
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000105851-PIK3CG/tissue/kidney#imid_20552755
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA069976; green = PIK3CG, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA069976; green = PIK3CG, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PIK3CG IHC & IF
🧪Start with breast tissue: glandular cells should show medium PIK3CG staining (HPA: breast glandular cells, Medium). Run adipose tissue as the negative, where adipocytes are not detected (HPA: adipose adipocytes, Not detected); on the breast slide, use non-glandular cells as an internal background comparator only if they remain unstained.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PIK3CG in HEL, HUVEC/TERT2, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality when known (selected tissue-IHC caption: rabbit primary). Use PIK3CG knockout tissue or a validated peptide-blocking control to assess target specificity; for the caption’s SABC/DAB detection, check endogenous biotin and peroxidase background (selected tissue-IHC caption: SABC/DAB).
⚠️Feasibility: A PIK3CG-specific fixation window or fixation effect is unreported, and the selected A01517-2 paraffin-section caption does not state a fixative (selected tissue-IHC caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected tissue-IHC caption). The supplied evidence does not establish whether frozen sections or IF are easier; membrane and cytosolic signal provide IF localization checks (HPA subcellular: plasma membrane and cytosol), while endogenous biotin or peroxidase can complicate interpretation of the caption’s SABC/DAB breast staining (selected tissue-IHC caption: SABC/DAB).
HPA tissue IHC evidence for PIK3CG
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | 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 → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PIK3CG IHC Tips
Troubleshoot PIK3CG staining by checking retrieval, compartment, cell identity and detection controls before assigning biological meaning to chromogenic signal.
Which retrieval conditions should I try first for PIK3CG in paraffin sections?
Start PIK3CG paraffin-section IHC with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A01517-2). The selected tissue image used that retrieval before overnight incubation at 4°C with 2 μg/mL catalog antibody (datasheet A01517-2). If staining is weak, first check that sections were fully deparaffinised and that heating and cooling were consistent across the run (standard IHC practice). Compare any retrieval adjustment on adjacent sections with the same antibody concentration and detection conditions, because excessive heating can damage morphology (standard IHC practice). Judge improvement by interpretable cytoplasmic or membranous staining, not by stronger background alone (HPA tissue IHC; standard IHC practice).
How should I assess fixation when PIK3CG staining is weak or uneven?
The selected image identifies a paraffin-embedded human breast cancer section, but does not report its fixative (datasheet A01517-2). Target-specific fixation sensitivity is therefore unknown; the image, HPA tissue patterns and PIK3CG topology do not establish a preferred fixative (datasheet A01517-2; HPA tissue IHC; UniProt P48736 topology). Record each specimen’s fixative and fixation duration, then compare similarly processed sections in the same staining run (standard IHC practice). Check whether weak regions follow tissue thickness, folds or uneven processing before changing antibody concentration (standard IHC practice). Retain morphology and a consistent positive control when comparing fixation conditions, so differences in signal remain interpretable (standard IHC practice).
Should PIK3CG staining be cytoplasmic, membranous or nuclear?
Expect cytoplasmic and membranous PIK3CG staining in tissue sections (HPA tissue IHC; UniProt P48736 subcellular location). Supported subcellular imaging places the protein mainly at the plasma membrane, with additional cytosolic localisation (HPA subcellular). PIK3CG has no transmembrane segment, so membrane-associated signal need not resemble the continuous outline of an integral membrane protein (UniProt P48736 topology; standard IHC interpretation). Assess whether staining follows cell boundaries or cytoplasm in intact cells, using the counterstain to identify nuclei and tissue structure (standard IHC practice). Predominantly nuclear DAB should prompt review of controls and morphology before being called PIK3CG (HPA tissue IHC; standard IHC practice).
Can this stain distinguish PIK3CG isoforms or modification states?
The supplied record lists 0 alternative isoforms for PIK3CG, but the selected image does not identify the catalog antibody’s epitope (UniProt P48736 isoforms; datasheet A01517-2). Its DAB pattern therefore cannot establish recognition of a particular domain or modification state (datasheet A01517-2; standard IHC interpretation). PIK3CG contains a catalytic region at residues 797–1080 and reported phosphorylation sites at 1024 and 1101 (UniProt P48736 domains and modified residues). Check epitope documentation before interpreting staining after epitope-altering treatment or comparing antibodies raised against different regions (standard IHC practice). Use an independent specificity control when a domain- or phosphorylation-specific conclusion matters (standard IHC practice).
How can IF help resolve which cells carry the PIK3CG signal?
Use IF as a separate localisation check alongside the chromogenic tissue readout, and multiplex PIK3CG with a marker such as CD45 when assessing hematopoietic cells (HPA tissue IHC; standard IF practice). The tissue profile includes medium PIK3CG staining in bone-marrow hematopoietic cells (HPA tissue IHC). Choose a fluorophore toward the red end of the spectrum when tissue autofluorescence obscures shorter-wavelength signal, and include single-channel controls (standard IF practice). If the antibody epitope faces the cytosol, optimise permeabilisation to expose it without losing membrane detail; PIK3CG lacks a transmembrane segment (UniProt P48736 topology; standard IF practice). Compare cell-associated signal with the supported plasma-membrane and cytosolic locations (HPA subcellular).
How do I separate PIK3CG signal from diffuse DAB background?
The selected IHC image used 10% goat-serum blocking, a biotinylated secondary antibody, a streptavidin–biotin complex and DAB (datasheet A01517-2). For diffuse staining, review blocking and washing, then compare a no-primary section processed through the same detection steps (standard IHC practice). Include a peroxidase block for DAB detection, and assess endogenous biotin when background tracks the biotin-based detection system (standard IHC practice; datasheet A01517-2 detection method). Keep DAB development and counterstaining consistent between control and test sections so intensity comparisons are meaningful (standard IHC practice). Treat signal in damaged tissue or at section edges cautiously, even when adjacent intact cells stain (standard IHC practice).
What should I score when comparing PIK3CG across IHC sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue PIK3CG staining can be cytoplasmic or membranous (HPA tissue IHC). For a defined population, report the percentage of positive cells and intensity, or calculate an H-score from intensity categories with a possible range of 0–300 (standard IHC scoring practice). When immune-cell abundance varies, report positive-cell density per mm² of viable tissue and normalise positive counts to the relevant cell population (standard IHC quantification practice; UniProt P48736 immune function). Exclude folds, necrosis and poorly preserved regions using consistent rules (standard IHC practice). Apply identical exposure-independent scoring criteria and detection conditions across sections (standard chromogenic IHC practice).
Which staining patterns warrant caution before calling a PIK3CG-positive sample?
A credible call places signal in intact cells with cytoplasmic or membranous distribution and fits the identified cell population (HPA tissue IHC; UniProt P48736 subcellular location). The HPA tissue profile reports medium staining in bone-marrow hematopoietic cells and no detected staining in heart-muscle cardiomyocytes, so cell identity matters more than the organ name alone (HPA tissue IHC). Predominantly nuclear staining, staining confined to necrosis or section edges, and signal reproduced without primary antibody warrant investigation before scoring (HPA tissue IHC; standard IHC practice). Check endogenous peroxidase and biotin-related background when using the selected DAB and biotin-based detection workflow (datasheet A01517-2; standard IHC practice). Interpret biological differences only after controls and tissue morphology support the staining pattern (standard IHC practice).
Boster reagents
Best PIK3CG / Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform IHC Antibodies
A01517-2 has real IHC data from a human paraffin-embedded breast cancer section (image caption). Catalog reactivity includes human, mouse, and rat (catalog: reactivity); no IF data are supplied (catalog: IF images).
Real IHC data
Anti-PI3K-gamma/PIK3CG Antibody ®
Cat # A01517-2
A01517-2 is listed for IHC and was shown on a human paraffin-embedded breast cancer section (catalog: applications; image caption). Its listed reactivity is human, mouse, and rat, while the supplied IHC image documents human tissue only (catalog: reactivity; image caption).
Which to pick: Choose A01517-2 for paraffin-section IHC: its image shows staining at 2 μg/ml after heat-mediated EDTA retrieval at pH 8.0 (image caption). For mouse or rat tissue, treat the listed reactivity as a starting point for validation; the supplied IHC image is human, the fixative is unreported, and clonality is unreported (catalog: reactivity; image caption; catalog: clone). No IF/ICC application or IF image is supplied for A01517-2, so this payload does not support an IF/ICC pick (catalog: applications; catalog: IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P48736 (PK3CG_HUMAN, Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform).
- Human Protein Atlas. PIK3CG tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PIK3CG subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the cytosol..
- Human Protein Atlas. PIK3CG antibody validation summary (1 antibodies).
- Revealing the Effects of the Herbal Pair of Euphorbia kansui and Glycyrrhiza on Hepatocellular Carcinoma Ascites with Integrating Network Target Analysis and Experimental Validation. International journal of biological sciences 2016 — PMC4852206.
- The potential therapeutic regimen for overcoming resistance to osimertinib due to rare mutations in NSCLC. iScience 2023 — PMC10320197.
- Inhibition of PI3K Isoform p110γ Increases Both Anti-Tumor and Immunosuppressive Responses to Aggressive Murine Head and Neck Squamous Cell Carcinoma with Low Immunogenicity. Cancers 2021 — PMC7956466.
- Repressing phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma by microRNA-142-3p restrains the progression of hepatocellular carcinoma. Bioengineered 2022 — PMC8805872.
- PubMed PMID:7624799 — UniProt-cited evidence.
- PubMed PMID:12853948 — UniProt-cited evidence.
- PubMed PMID:12690205 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PIK3CG (Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit gamma isoform). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01517-2). Validated IHC antibody: A01517-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A01517-2); 2. Fixation: Image fixative and duration unreported (datasheet A01517-2); 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 A01517-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01517-2); 8. Primary antibody: Rabbit anti-PIK3CG, 2-5μg/ml (datasheet A01517-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01517-2). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding may confound staining (HPA tissue IHC); Isoform / epitope: 0 listed isoforms; one full-length intracellular chain (UniProt); Regulation: Stimulus-dependent expression is unreported (UniProt). Evidence: UniProt P48736, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A01517-2, PMC8805872. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PIK3CG/immunohistochemistry-ihc-guide.