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PCYOX1 / Prenylcysteine oxidase 1 · IHC design guide
Design Immunohistochemistry for PCYOX1
Plan paraffin-section PCYOX1 IHC around cytoplasmic staining in most tissues, including high staining in hepatocytes and kidney tubule cells (HPA tissue IHC). Use the catalog antibody’s IHC-P conditions (datasheet A11180-1), and interpret staining with HPA’s reported low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PCYOX1 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 tissue staining (HPA tissue IHC); lysosomal protein (UniProt) | |
| Staining pattern | Cytoplasmic staining in many cell types across most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A11180-1) | |
| Positive control | Heart muscle+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11180-1) | |
| Caveat | Staining may diverge from RNA expression (HPA tissue IHC) | |
| Regulation | Widely expressed; no regulator specified (UniProt) | |
| Isoform / epitope | Two isoforms; signal peptide cleaved; epitope impact unknown (UniProt) |
Section 1
Recommended PCYOX1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with a published protocol for human atherosclerotic lesions (PMC8455616).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A11180-1) |
| Fixation | Image fixative and duration unreported (datasheet A11180-1); 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 A11180-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A11180-1) |
| Primary antibody | Rabbit anti-PCYOX1, 2-5 μg/ml (datasheet A11180-1) |
| Primary incubation | Overnight at 4 °C (datasheet A11180-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A11180-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PCYOX1-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A11180-1). Use citrate pH 6.0 to reproduce the published protocol (PMC8455616).
Section 2
What Is the Expected PCYOX1 Staining Pattern?
PCYOX1 is lysosomal and has no transmembrane segment (UniProt Q9UHG3). In paraffin sections, expect cytoplasmic staining in the relevant cells, including hepatocytes, kidney tubule cells and cardiomyocytes, which stain High in HPA tissue IHC (HPA: tissue IHC). HPA describes cytoplasmic expression in most tissues but rates the staining Approved with low consistency between antibody staining and RNA expression; interpret unexpected patterns cautiously (HPA: tissue IHC).
| Cytoplasmic signal in hepatocytes, kidney tubule cells or cardiomyocytes, with visible cell boundaries. | These are useful positive reference populations: each is rated High in HPA tissue IHC (HPA: liver, kidney and heart muscle). A lysosomal protein can give intracellular staining (UniProt Q9UHG3); HPA reports a broad cytoplasmic pattern and does not establish that every positive cell must show discrete puncta (HPA: tissue IHC). |
| Predominantly nuclear staining or a sharp outline confined to the plasma membrane. | That distribution does not match the reported cytoplasmic tissue pattern or lysosomal location (HPA: tissue IHC; UniProt Q9UHG3). Treat it as suspect until it is reproduced with a suitable control. A compartment mismatch alone cannot identify whether the cause is nonspecific binding, detection background or an interpretation error. |
| Strong signal in hematopoietic cells of bone marrow or germinal center cells of tonsil. | Those particular cell populations are Not detected in HPA tissue IHC (HPA: bone marrow; HPA: tonsil). Check cell identity and a no-primary control. Persistent staining could reflect antibody cross-reactivity or endogenous detection activity; a discordant slide alone cannot establish which explanation applies. |
| Diffuse chromogen across cells and extracellular spaces, obscuring intracellular detail. | The result is hard to score as PCYOX1 because HPA describes cytoplasmic expression in most tissues (HPA: tissue IHC). General IHC practice: compare a no-primary control and background in adjacent tissue, then review blocking, washes and detection chemistry. Widespread color without recognizable cell-specific staining is insufficient evidence of a positive result. |
| No visible signal in hepatocytes, kidney tubule cells or cardiomyocytes. | Absence in one of these High populations conflicts with the HPA reference pattern (HPA: liver, kidney and heart muscle). First confirm that the expected cell population is present and that the run's positive control worked. If both checks pass, review the antibody and detection workflow before calling the specimen negative. |
💡Expected PCYOX1 appearanceCall a result positive when cytoplasmic staining is clear in an expected cell population, such as High-staining hepatocytes or kidney tubule cells (HPA: liver; HPA: kidney); dominant nuclear or membrane-only color is discordant with the reported cytoplasmic pattern and lysosomal location (HPA: tissue IHC; UniProt Q9UHG3).
| Cell population and tissue | PCYOX1 is widely expressed (UniProt Q9UHG3), yet HPA levels vary by cell population: Leydig cells are High, while ovarian stroma cells are Not detected (HPA: testis; HPA: ovary). Score the identified cells, not the whole section. |
| Antibody evidence | HPA035193 is a rabbit polyclonal antibody with Approved IHC status (HPA: antibodies). HPA also reports low consistency between staining and RNA expression (HPA: tissue IHC), so the rating is not independent proof of every observed signal. |
| Protein processing and glycosylation | The annotated signal peptide spans residues 1–27; the mature chain spans 28–505, with three glycosylation sites (UniProt Q9UHG3). Epitope position is unspecified here, so these annotations do not predict retrieval needs or staining intensity. |
| Isoforms | Two isoforms are listed (UniProt Q9UHG3). The supplied HPA IHC observations do not distinguish them (HPA: tissue IHC); an IHC-positive cell therefore cannot be assigned to isoform 1 or 2 from these data. |
| IF/ICC Q&A: Is there a confirmed cellular image pattern? | No HPA ICC-IF image, main location or ICC validation entry is supplied (HPA: subcellular; HPA: antibodies). Lysosomal localization is an expectation from UniProt Q9UHG3, not an HPA-confirmed IF pattern; use the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue is blank. | The reference cell type may be absent, or the staining run may have failed; hepatocytes, kidney tubule cells and cardiomyocytes are High references (HPA: tissue IHC). | Verify the cells on the section and compare an independent positive control. General IHC practice: check reagent sequence, detection and the validated retrieval conditions before interpreting a negative result. |
| Signal is much weaker than the reference image. | HPA's High, Medium and Not detected calls apply to specified cell populations (HPA: tissue IHC). A different population or scoring threshold can make a section appear weaker. | Compare like cells at similar magnification and document the proportion and intensity of stained cells. General IHC practice: assess the run control before changing staining conditions. |
| Nuclear or membrane-only color dominates. | This conflicts with HPA's cytoplasmic tissue pattern and UniProt's lysosomal location (HPA: tissue IHC; UniProt Q9UHG3). The supplied sources do not identify a PCYOX1-specific cause. | Check the no-primary control, tissue morphology and antibody localization in a High reference population (HPA: tissue IHC). Reassess the pattern only after background has been excluded. |
| Bone marrow hematopoietic cells or tonsil germinal centers stain strongly. | Both populations are Not detected in HPA tissue IHC (HPA: bone marrow; HPA: tonsil). Cross-reactivity or endogenous chromogenic activity is possible, but neither is established by the slide alone. | Confirm the cell type and compare a no-primary control. General IHC practice: if enzyme-based detection is used, check the appropriate endogenous-activity control. |
| Color covers most of the section and masks cell detail. | Diffuse background can obscure the cytoplasmic pattern reported by HPA (HPA: tissue IHC). The supplied evidence does not establish a target-specific fixation or retrieval effect. | General IHC practice: inspect a no-primary control, blocking, washes and detection exposure; then repeat with a positive reference tissue in the same run. |
| An IF/ICC image seems to show a different compartment. | HPA supplies no ICC-IF images or main subcellular location for PCYOX1 (HPA: subcellular). UniProt assigns PCYOX1 to lysosomes (UniProt Q9UHG3), but that does not validate the image. | Treat the image as unconfirmed for IF/ICC and consult that application's separate guide. Do not transfer an IHC-P staining pattern into an IF/ICC validation claim. |
Experiment evidence
IHC & IF images showing PCYOX1 staining
A11180-1 · IHCIHC analysis of PCYOX1 using anti-PCYOX1 antibody (A11180-1). PCYOX1 was detected in a paraffin-embedded section of human breast cancer tiss
A11180-1 · IHCIHC analysis of PCYOX1 using anti-PCYOX1 antibody (A11180-1). PCYOX1 was detected in a paraffin-embedded section of human breast cancer tiss
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000116005-PCYOX1/tissue/cerebral+cortex#imid_8370388
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000116005-PCYOX1/tissue/colon#imid_8370314
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000116005-PCYOX1/tissue/liver#imid_8370307
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000116005-PCYOX1/tissue/kidney#imid_8370308
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PCYOX1 IHC & IF
🧪Run kidney first: cells in tubules must stain (HPA: High in kidney tubules). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in hematopoietic cells); the supplied rows do not identify a validated internal-negative population in kidney, so use only independently verified negative cells on that slide, which should show counterstain without specific DAB signal (HPA: kidney and bone marrow rows; standard IHC practice).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PCYOX1; derive a cell-line control from the positive tissue's cell type (Cardiomyocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; a concentration-matched rabbit IgG isotype control matched to the primary’s class and clonality when known; and PCYOX1-knockout material as a biological negative (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase in kidney sections before HRP/DAB detection and assess residual tissue pigment against the controls (caption: HRP/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11180-1 paraffin-section caption does not state the fixative (caption: fixative unreported). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required; frozen sections and IF/ICC have no comparative feasibility evidence in the supplied payload (caption: EDTA retrieval; HPA: no ICC-IF images listed). For kidney, assess endogenous peroxidase and tissue pigment when interpreting DAB staining (caption: HRP/DAB; standard IHC practice).
HPA tissue IHC evidence for PCYOX1
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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Liver | Hepatocytes | High | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PCYOX1 IHC Tips
Troubleshoot PCYOX1 staining by checking retrieval, cellular pattern, and controls before scoring signal (datasheet A11180-1; UniProt Q9UHG3; standard IHC practice).
How should I retrieve PCYOX1 antigen in paraffin sections with weak staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections stained with A11180-1 (datasheet A11180-1). Keep heating and cooling conditions consistent across comparison slides, because retrieval variation can alter chromogenic intensity (standard IHC practice). The selected image used 2 μg/mL primary antibody overnight at 4°C, so first check whether those incubation conditions were followed before changing retrieval chemistry (datasheet A11180-1). If staining remains weak, compare a small retrieval time series on adjacent sections while monitoring tissue integrity (standard IHC practice). Record the chosen heating duration and compare it against a positive control processed in the same run (standard IHC practice).
Could fixation explain weak or uneven PCYOX1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A11180-1). Record the actual fixative, fixation duration, section age, and processing history for each specimen before attributing signal differences to PCYOX1 biology (standard IHC practice). Compare sections processed under matched conditions, with a known positive tissue on the same run, and check whether weak areas coincide with poor morphology or uneven reagent access (standard IHC practice; HPA: high in kidney tubule cells). If fixation conditions differ, treat staining intensity as potentially confounded until matched specimens or an independent assay support the comparison (standard IHC practice).
What staining pattern is plausible for PCYOX1 in tissue?
Expect intracellular staining compatible with a lysosomal protein, although chromogenic IHC may resolve it only as cytoplasmic signal (UniProt Q9UHG3: lysosome; HPA: cytoplasmic expression in most tissues; standard IHC practice). PCYOX1 has no annotated transmembrane segment, so a sharp plasma-membrane outline would need independent validation rather than automatic acceptance (UniProt Q9UHG3 topology). Compare the pattern within identifiable cell populations: HPA reports high staining in hepatocytes and kidney tubule cells, but no detection in bone-marrow hematopoietic cells (HPA tissue IHC). Use morphology and a negative control to distinguish granular intracellular signal from diffuse chromogen deposits (standard IHC practice).
Can processing or isoforms change what this antibody detects?
PCYOX1 has 2 annotated isoforms and a signal peptide spanning residues 1–27; the annotated mature chain spans 28–505 (UniProt Q9UHG3). Its 3 annotated glycosylation sites are residues 196, 323, and 353, but the supplied evidence does not map this antibody's epitope (UniProt Q9UHG3; datasheet A11180-1). Consequently, do not assign a weak or absent tissue stain to a particular isoform, cleavage event, or glycosylation state without epitope mapping and independent validation (standard IHC practice). When comparing specimens, keep retrieval and detection conditions matched so technical differences do not masquerade as epitope-dependent biology (standard IHC practice).
How can I assess PCYOX1 by IF alongside a cell-type marker?
Treat IF as a separate optimisation: the selected A11180-1 evidence describes chromogenic staining of a paraffin section, while HPA supplies no PCYOX1 ICC/IF images (datasheet A11180-1; HPA subcellular). Multiplex PCYOX1 with a marker for the cell population being examined, such as hepatocytes in liver or tubule cells in kidney, and check each channel separately before judging overlap (HPA tissue IHC; standard IF practice). Select fluorophores after measuring tissue autofluorescence, favouring a brighter, spectrally separated channel for a weak target (standard IF practice). Because PCYOX1 is lysosomal but the antibody epitope's accessibility is unspecified, titrate permeabilisation and verify access empirically rather than assuming which side of a membrane contains the epitope (UniProt Q9UHG3; standard IF practice).
How do I reduce diffuse or misleading DAB background?
The selected tissue image used 10% goat serum blocking, 2 μg/mL primary antibody overnight at 4°C, and an anti-rabbit peroxidase secondary for 30 minutes at 37°C (datasheet A11180-1). For diffuse background, compare a primary-antibody titration with matched wash and DAB development times, while retaining an on-run positive control (standard IHC practice; HPA: high in hepatocytes). Include a no-primary control to reveal secondary-reagent binding or endogenous peroxidase contribution; apply a peroxidase block as a general chromogenic IHC step (standard IHC practice). Inspect precipitate outside cells and at section edges before interpreting granular cytoplasmic colour as PCYOX1 (standard IHC practice; HPA: cytoplasmic expression).
How should I score PCYOX1 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Choose a prespecified cell population and report the percentage of positive cells plus intensity, or calculate an H-score from intensity categories 0–3 for a possible range of 0–300 (standard IHC practice). Normalise counts to the number of eligible cells, or report positive-cell density per mm² of viable tissue when cell counting is impractical (standard IHC practice). Score matched anatomical regions and exclude necrosis, folds, and section edges using rules fixed before review (standard IHC practice). Keep staining runs and image exposure comparable, and report the cell compartment assessed because HPA describes PCYOX1 staining as cytoplasmic in most tissues (standard IHC practice; HPA tissue IHC).
When is a PCYOX1-positive stain convincing rather than artefactual?
A convincing result shows reproducible intracellular signal in morphologically identifiable cells, with little signal in the no-primary control (UniProt Q9UHG3: lysosome; HPA: cytoplasmic expression; standard IHC practice). HPA reports high staining in kidney tubule cells and hepatocytes, yet labels its tissue IHC reliability Approved with low consistency between staining and RNA expression, so those patterns are useful checks rather than definitive proof (HPA tissue IHC). Question a predominantly nuclear or crisp plasma-membrane pattern, edge-only colour, necrotic deposits, or staining explained by endogenous peroxidase (UniProt Q9UHG3 topology; standard IHC practice). For unexpected cell populations, confirm the observation with an independent method before assigning it to PCYOX1 (standard IHC practice).
Boster reagents
Best PCYOX1 / Prenylcysteine oxidase 1 IHC Antibodies
A11180-1 has real IHC images from human paraffin sections of breast cancer and testicular seminoma (A11180-1 image captions). No IF/ICC data are listed (catalog applications and image payload).
Real IHC data
Anti-PCYOX1 Antibody ®
Cat # A11180-1
A11180-1 is listed for human IHC and is shown staining paraffin sections of human breast cancer (catalog applications and reactivity; A11180-1 image captions). The same SKU is shown staining paraffin sections of human testicular seminoma (A11180-1 image captions).
Which to pick: For human paraffin-section IHC, choose A11180-1: its captions document EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (A11180-1 image captions), within the listed 2–5 μg/ml IHC range (catalog dilution). No listed SKU has IF/ICC validation or an IF image (catalog applications and image payload). No cross-species choice is supported because A11180-1 lists human reactivity only; its captions specify paraffin sections but do not report the fixative (catalog reactivity; A11180-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UHG3 (PCYOX_HUMAN, Prenylcysteine oxidase 1).
- Human Protein Atlas. PCYOX1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PCYOX1 subcellular location (ICC-IF): Highest expression in ASC52telo: 149.3 nTPM.
- Human Protein Atlas. PCYOX1 antibody validation summary (1 antibodies).
- Prenylcysteine oxidase 1, an emerging player in atherosclerosis. Communications biology 2021 — PMC8455616.
- PubMed PMID:10585463 — UniProt-cited evidence.
- PubMed PMID:10048485 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PCYOX1 (Prenylcysteine oxidase 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); lysosomal protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A11180-1). Validated IHC antibody: A11180-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A11180-1); 2. Fixation: Image fixative and duration unreported (datasheet A11180-1); 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 A11180-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A11180-1); 8. Primary antibody: Rabbit anti-PCYOX1, 2-5 μg/ml (datasheet A11180-1); 9. Primary incubation: Overnight at 4 °C (datasheet A11180-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11180-1); Caveat: Staining may diverge from RNA expression (HPA tissue IHC); Isoform / epitope: Two isoforms; signal peptide cleaved; epitope impact unknown (UniProt); Regulation: Widely expressed; no regulator specified (UniProt). Evidence: UniProt Q9UHG3, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in ASC52telo: 149.3 nTPM, Datasheet A11180-1, PMC8455616. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PCYOX1/immunohistochemistry-ihc-guide.