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PYCR1 / Pyrroline-5-carboxylate reductase 1, mitochondrial · IHC design guide
Design Immunohistochemistry for PYCR1
Plan PYCR1 staining in paraffin sections using cytoplasmic tissue staining as the observed pattern (HPA tissue IHC) and mitochondrial location as the molecular expectation (UniProt). The guide covers fixation, controls and interpretation, including the reported low consistency between staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PYCR1 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 staining (HPA tissue IHC); mitochondrial location (UniProt) | |
| Staining pattern | Cytoplasmic in most tissues; high in select glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06018-1) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Oral mucosa+2 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 and RNA expression have low consistency (HPA tissue IHC) | |
| Regulation | Pancreas and salivary gland RNA enhanced (HPA RNA) | |
| Isoform / epitope | 3 isoforms; epitope impact is unknown (UniProt) |
Section 1
Recommended PYCR1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with four published PYCR1 staining workflows for renal, osteosarcoma, esophageal, and pancreatic tissues (PMC6641676; PMC13219957; PMC9218025; PMC8965111).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A06018-1) |
| Fixation | Image fixative and duration unreported (datasheet A06018-1); 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 A06018-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06018-1) |
| Primary antibody | Rabbit anti-PYCR1, 2-5 μg/ml (datasheet A06018-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06018-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06018-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PYCR1-positive staining in basal cells of bronchus (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 retrieval at pH 8.0 for the catalog antibody (datasheet A06018-1); published citrate protocols used different antibodies (PMC6641676; PMC13219957; PMC8965111).
Section 2
What Is the Expected PYCR1 Staining Pattern?
PYCR1 localises to mitochondria and has no annotated transmembrane segment (UniProt P32322: location and topology). In paraffin-section IHC, expect cytoplasmic staining in many tissues, including strong staining in pancreatic exocrine glandular cells and bronchial basal cells (HPA: tissue IHC). HPA rates its antibody staining Approved, while reporting low consistency with RNA expression; interpret the pattern alongside controls rather than treating intensity alone as proof of specificity (HPA: reliability).
| Cytoplasmic signal in pancreatic exocrine glandular cells, with little signal in ovarian stroma. | This fits a useful positive–negative comparison: HPA reports High staining in pancreatic exocrine glandular cells and Not detected in ovarian stroma cells (HPA: tissue IHC). A granular cytoplasmic appearance can be compatible with mitochondrial localisation, but chromogenic IHC alone does not resolve individual mitochondria (UniProt P32322: location; general IHC practice). |
| Predominantly nuclear or surface-restricted signal, with little cytoplasmic staining. | That distribution conflicts with the reported mitochondrial location and HPA’s cytoplasmic tissue profile (UniProt P32322: location and topology; HPA: tissue IHC). Check the counterstain and detection controls before assigning it to PYCR1; the pattern could reflect artefact or another antigen (general IHC practice). |
| Strong staining in ovarian stroma cells or oral squamous epithelial cells. | HPA reports PYCR1 as Not detected in those cell populations (HPA: tissue IHC). Reproducible staining there warrants a specificity check, including a no-primary control; cross-reactivity or endogenous detection activity are possibilities, not diagnoses from appearance alone (general IHC practice). |
| Broad, diffuse colour across cells and surrounding tissue without clear cell boundaries. | This is difficult to score as the reported cell-associated cytoplasmic pattern (HPA: tissue IHC). Compare a no-primary control and review blocking, washes and chromogen development for background from the detection workflow (general IHC practice). |
| No signal in pancreatic exocrine glandular cells while the section is otherwise interpretable. | An expected strong reference population is missing (HPA: High in pancreatic exocrine glandular cells). First verify tissue identity and assay controls, then review the antibody’s IHC-P instructions and retrieval and detection steps; a blank slide alone cannot establish absent PYCR1 (general IHC practice). |
💡Expected PYCR1 appearanceCall a positive result when cell-associated cytoplasmic staining is clear in an HPA High population such as pancreatic exocrine glandular cells; isolated nuclear or diffuse tissue-wide colour is suspect (HPA: tissue IHC; UniProt P32322: location; general IHC practice).
| Compartment and optical resolution | PYCR1 is mitochondrial and lacks an annotated transmembrane segment (UniProt P32322: location and topology). HPA describes cytoplasmic tissue staining; a chromogenic section can support that compartment call without proving mitochondrial colocalisation (HPA: tissue IHC; general IHC practice). |
| Reference cell populations | HPA reports High staining in bronchial basal cells, cortical and hippocampal neuronal cells, and pancreatic exocrine glandular cells; oral squamous epithelial cells, ovarian stroma cells and soft-tissue fibroblasts are Not detected (HPA: tissue IHC). Score the named cells within each tissue, rather than the whole section. |
| Strength of antibody evidence | The listed antibody HPA047660 has Approved IHC status; HPA also flags low agreement between antibody staining and RNA expression (HPA: antibody and tissue records). This supports cautious use of the observed pattern, with controls, rather than treating every coloured cell as confirmed PYCR1. |
| Isoforms and processing | UniProt lists three PYCR1 isoforms and a chain spanning residues 2–319 (UniProt P32322: isoforms and processing). These annotations do not identify the catalog antibody’s epitope or establish different staining among isoforms; avoid inferring either from an IHC intensity difference. |
| IF/ICC: what pattern is established? | Mitochondrial localisation predicts intracellular signal (UniProt P32322: location), but HPA supplies no main ICC-IF location or cell-line images, and the listed antibody has no ICC status (HPA: subcellular and antibody records). An IF result needs its own validation; no IF protocol is established here. |
| Situation | Likely cause | Next action |
|---|---|---|
| Pancreatic exocrine glandular cells remain blank. | A reference population reported High by HPA is missing; the slide alone cannot distinguish assay failure from sample variation (HPA: tissue IHC; general IHC practice). | Confirm cell identity and control performance, then check the catalog antibody’s IHC-P instructions, retrieval, dilution and detection steps (general IHC practice). |
| Nuclei dominate the stain. | A nuclear-dominant result conflicts with mitochondrial location and the reported cytoplasmic tissue profile (UniProt P32322: location; HPA: tissue IHC). | Compare the counterstain and no-primary control; review whether chromogen development or nonspecific binding obscures the cytoplasm (general IHC practice). |
| Oral squamous epithelium stains strongly. | HPA reports this cell population as Not detected; cross-reactivity or endogenous detection activity could explain an unexpected signal (HPA: tissue IHC; general IHC practice). | Run a no-primary control and review blocking and detection controls before scoring that staining as PYCR1 (general IHC practice). |
| Colour covers tissue and empty spaces evenly. | A field-wide deposit is inconsistent with a cell-associated cytoplasmic pattern (HPA: tissue IHC); excess detection background is possible (general IHC practice). | Inspect the no-primary control, washing and chromogen development, then rescore only clearly localised cellular signal (general IHC practice). |
| All cell types appear equally intense. | HPA reports different levels across specified cell populations, from High to Not detected (HPA: tissue IHC). Uniform colour can make those differences unreadable (general IHC practice). | Check exposure to chromogen and compare named positive and negative populations on controlled sections before assigning intensity categories (general IHC practice). |
| A granular pattern is hard to distinguish from background. | Mitochondrial location is annotated, while HPA reports tissue staining at the broader cytoplasmic level (UniProt P32322: location; HPA: tissue IHC). | Score reproducible cellular cytoplasmic staining against controls; reserve a mitochondrial colocalisation claim for separately validated IF evidence (general IHC practice). |
Experiment evidence
IHC & IF images showing PYCR1 staining
A06018-1 · IHCIHC analysis of PYCR1 using anti-PYCR1 antibody (A06018-1). PYCR1 was detected in a paraffin-embedded section of human lung cancer tissue. H
A06018-1 · IHCIHC analysis of PYCR1 using anti-PYCR1 antibody (A06018-1). PYCR1 was detected in a paraffin-embedded section of human ovarian cancer tissue
A06018-1 · IFIF analysis of PYCR1 using anti-PYCR1 antibody (A06018-1). PYCR1 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000183010-PYCR1/tissue/cerebral+cortex#imid_13165575
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000183010-PYCR1/tissue/colon#imid_13165509
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000183010-PYCR1/tissue/liver#imid_13165510
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000183010-PYCR1/tissue/kidney#imid_13165511
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PYCR1 IHC & IF
🧪Run bronchus first and score staining in basal cells, which HPA rates High (HPA: bronchus basal cells, High). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected); non-basal cells on the bronchus slide can be compared with background, but HPA does not establish that they are PYCR1-negative.
Positive control tissue: Bronchus (Basal cells, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PYCR1; derive a cell-line control from the positive tissue's cell type (Basal cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, matching the primary antibody’s clonality where known (caption: rabbit anti-PYCR1); use a PYCR1 knockout sample as a biological negative if available. Block endogenous peroxidase and check inflammatory cells for residual chromogen signal in bronchus sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the catalog antibody in a paraffin-embedded lung cancer section, but its dependence on that retrieval condition in bronchus is unreported (caption: A06018-1). The supplied evidence does not establish that frozen sections or IF are easier; endogenous peroxidase in bronchial inflammatory cells can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for PYCR1
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 |
|---|---|---|---|---|
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced PYCR1 IHC Tips
Troubleshoot PYCR1 staining in paraffin sections by checking retrieval, controls, cellular distribution and scoring before interpreting chromogenic signal.
Which retrieval conditions should I try first for PYCR1 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06018-1). The selected IHC image used this condition before incubation with 2 μg/ml primary antibody overnight at 4°C (datasheet A06018-1). If staining is weak, check that heating and cooling were consistent across sections before testing another retrieval condition (standard IHC practice). Compare candidate conditions on adjacent sections with the same antibody concentration, detection reagents and exposure to DAB (standard IHC practice). Judge improvement by cellular staining and preserved morphology, not by stronger background alone (standard IHC practice).
Could fixation explain weak or uneven PYCR1 staining?
The selected image identifies a paraffin-embedded section but does not state its fixative, so PYCR1-specific fixation sensitivity is unknown (datasheet A06018-1). Record the specimen's fixative and fixation interval, and compare sections processed together when investigating weak staining (standard IHC practice). Examine morphology and staining at section edges before changing retrieval, because uneven processing can confound a comparison (standard IHC practice). Keep EDTA at pH 8.0, antibody at 2 μg/ml and overnight incubation at 4°C constant during that comparison (datasheet A06018-1). Do not infer a fixation preference from PYCR1 localisation or tissue staining patterns (UniProt P32322; HPA tissue IHC).
Where should convincing PYCR1 staining appear in chromogenic IHC?
Expect intracellular staining compatible with mitochondrial PYCR1, while recognising that light-microscope DAB signal may appear broadly cytoplasmic (UniProt P32322 subcellular location; HPA tissue IHC profile). The protein has no annotated transmembrane segment, so a crisp cell-surface outline needs independent validation before being called PYCR1 (UniProt P32322 topology). Compare the stained cells with section morphology and a known positive compartment, such as pancreatic exocrine glandular cells (HPA: High in pancreatic exocrine glandular cells). Include a no-primary control to assess detection-system signal (standard IHC practice). Document diffuse cytoplasmic, granular and nuclear patterns separately rather than merging them into one positive score (standard IHC practice).
How can isoforms or epitope accessibility complicate PYCR1 IHC?
PYCR1 has 3 annotated isoforms, but the supplied product image does not identify its antibody epitope or establish isoform coverage (UniProt P32322 isoforms; datasheet A06018-1). Its annotated chain spans residues 2–319, with N-acetylserine at residue 2 and phosphoserines at 278 and 301 (UniProt P32322 processing and modified residues). Do not attribute a staining difference to an isoform or modification without epitope mapping and a suitable validation control (standard IHC practice). First compare adjacent sections under identical EDTA pH 8.0 retrieval and detection conditions (datasheet A06018-1). If patterns still disagree, report the antibody and conditions alongside the observed cellular distribution (standard IHC practice).
How should I assess PYCR1 in a multiplex IF experiment?
Treat IF as a separate assay: the supplied antibody evidence is from paraffin-section chromogenic IHC, with no IF image or IF fixation condition supplied (datasheet A06018-1; HPA subcellular data). Multiplex PYCR1 with a validated exocrine glandular-cell marker when examining pancreas, where those cells show high tissue-IHC staining (HPA: High in pancreatic exocrine glandular cells). Select spectrally separated fluorophores, check unstained tissue for autofluorescence and place a weaker signal in a cleaner channel (standard IF practice). Permeabilise sufficiently for an intracellular mitochondrial antigen, while recognising that the epitope's exact membrane-facing side is unspecified (UniProt P32322 subcellular location and topology). Optimise IF fixation and permeabilisation with IF controls rather than importing IHC conditions (standard IF practice).
What should I check when PYCR1 DAB staining is widespread?
Check a no-primary section for signal from the detection system and inspect unstained tissue for pigment that could mimic DAB (standard IHC practice). Include an endogenous peroxidase block in the chromogenic workflow, then assess whether background falls without losing cellular staining (standard IHC practice). The selected image used 10% goat serum block and peroxidase-linked detection with DAB; these are reported image conditions, not proof that every dark deposit is PYCR1 (datasheet A06018-1). Compare antibody dilution or incubation changes against the reported 2 μg/ml overnight at 4°C condition (datasheet A06018-1). Score only signal with plausible cellular distribution and adequate morphology (UniProt P32322 subcellular location; standard IHC practice).
How should PYCR1 staining be scored across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then record the percentage of positive cells and staining intensity separately (standard IHC practice). An H-score can combine intensity categories with their positive-cell percentages; report the scoring rules and blinded review procedure (standard IHC practice). For spatial comparisons, positive-cell density per mm² can supplement the H-score when the analysed tissue area is measured (standard IHC practice). Normalise cell counts to eligible cells or area within the same compartment, and exclude folds, necrosis and section edges by a stated rule (standard IHC practice). Keep retrieval at EDTA pH 8.0 and detection settings consistent across compared slides (datasheet A06018-1; standard IHC practice).
How do I distinguish credible PYCR1 staining from artefact?
A credible pattern places signal in intact cells with intracellular distribution compatible with mitochondrial PYCR1 and the cytoplasmic tissue-IHC profile (UniProt P32322 subcellular location; HPA tissue IHC profile). Compare expected positive cells, such as pancreatic exocrine glandular cells, with a low or undetected compartment such as ovarian stroma (HPA: High in pancreatic exocrine glandular cells; Not detected in ovarian stroma). Investigate predominantly nuclear or cell-surface staining, edge-only staining, necrotic deposits and no-primary signal before calling a section positive (UniProt P32322 subcellular location; standard IHC practice). Endogenous enzyme activity can also produce chromogenic signal, so assess the peroxidase-block control (standard IHC practice). Interpret tissue differences cautiously because HPA reports low consistency between antibody staining and RNA expression (HPA reliability description).
Boster reagents
Best PYCR1 / Pyrroline-5-carboxylate reductase 1, mitochondrial IHC Antibodies
A06018-1 has IHC data from human lung and ovarian cancer paraffin sections and IF data from U2OS cells and human glioma paraffin sections (catalog image captions).
Real IHC data
Anti-PYCR1 Antibody ®
Cat # A06018-1
A06018-1 is listed for human IHC and IF (catalog applications and reactivity). Its captions show IHC in human lung and ovarian cancer paraffin sections, plus IF in U2OS cells and human glioma paraffin sections (catalog image captions).
Which to pick: Choose A06018-1 for human tissue IHC on paraffin sections; its IHC captions document those sections, while the fixative is unreported (catalog IHC image captions). For IF/ICC, A06018-1 is listed for both applications and has IF images from U2OS cells and human glioma paraffin sections (catalog applications and IF image captions). No cross-species choice is supported: A06018-1 lists human reactivity only, and its clonality is unreported (catalog reactivity and clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P32322 (P5CR1_HUMAN, Pyrroline-5-carboxylate reductase 1, mitochondrial).
- Human Protein Atlas. PYCR1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PYCR1 subcellular location (ICC-IF): Highest expression in Hep-G2: 475.1 nTPM.
- Human Protein Atlas. PYCR1 antibody validation summary (1 antibodies).
- The clinical significance of PYCR1 expression in renal cell carcinoma. Medicine 2019 — PMC6641676.
- Machine Learning-Based Identification and Validation of PYCR1 and PYGM as Prognostic Biomarkers for Osteosarcoma. Clinical Medicine Insights. Oncology 2026 — PMC13219957.
- Circ_0000705 facilitates proline metabolism of esophageal squamous cell carcinoma cells by targeting miR-621/PYCR1 axis. Discover oncology 2022 — PMC9218025.
- PYCR1: A Potential Prognostic Biomarker in Pancreatic Ductal Adenocarcinoma. Journal of Cancer 2022 — PMC8965111.
- PubMed PMID:1730675 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PYCR1 (Pyrroline-5-carboxylate reductase 1, mitochondrial). Expected localisation Cytoplasmic staining (HPA tissue IHC); mitochondrial location (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06018-1). Validated IHC antibody: A06018-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Basal cells); negative Oral mucosa. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A06018-1); 2. Fixation: Image fixative and duration unreported (datasheet A06018-1); 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 A06018-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06018-1); 8. Primary antibody: Rabbit anti-PYCR1, 2-5 μg/ml (datasheet A06018-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06018-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression have low consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope impact is unknown (UniProt); Regulation: Pancreas and salivary gland RNA enhanced (HPA RNA). Evidence: UniProt P32322, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in Hep-G2: 475.1 nTPM, Datasheet A06018-1, PMC6641676, PMC13219957. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PYCR1/immunohistochemistry-ihc-guide.