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PRDX6 / Peroxiredoxin-6 · IHC design guide
Design Immunohistochemistry for PRDX6
This guide uses PRDX6’s widespread cytoplasmic tissue staining (HPA tissue IHC) to plan paraffin-section controls and scoring. It covers fixation consistency, the catalog antibody’s 0.5–1 μg/mL IHC range (datasheet: PB9350), and chromogenic detection.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRDX6 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 in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet PB9350) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); optimize locally. | |
| Caveat | Urothelial cells show low staining (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; chain spans residues 2–224 (UniProt) |
Section 1
Recommended PRDX6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: PB9350) is accompanied by published PRDX6 IHC methods from a rat myocardial infarction study (PMC3459299) and human abdominal aortic aneurysm tissue (PMC5146935).
| Sample | Paraffin-embedded Human Mammary Cancer tissues; fixative not specified (datasheet PB9350) |
| Fixation | Image fixative and duration unreported (datasheet PB9350); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet PB9350) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9350) |
| Primary antibody | Rabbit anti-PRDX6, 0.5-1μg/ml (datasheet PB9350) |
| Primary incubation | Overnight at 4 °C (datasheet PB9350) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9350) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRDX6-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteTry heat-mediated citrate pH 6 retrieval first (datasheet: PB9350); neither published excerpt specifies a retrieval method (PMC3459299; PMC5146935).
Section 2
What Is the Expected PRDX6 Staining Pattern?
PRDX6 should appear mainly in the cytoplasm across many tissues (HPA tissue IHC: ubiquitous cytoplasmic expression; reliability Supported, with medium consistency between staining and RNA data). High staining is reported in adipocytes, bronchial respiratory epithelial cells, and bone marrow hematopoietic cells (HPA tissue IHC: High). A cytoplasmic pattern also fits its cytoplasm and lysosome annotations and lack of a transmembrane segment (UniProt P30041: subcellular location; topology).
| Clear cytoplasmic staining in adipocytes or bronchial respiratory epithelial cells. | This matches reported high staining in those cells (HPA tissue IHC: High). Judge signal within identified cells against nearby background; tissue-wide intensity alone can hide cell-level differences (standard IHC practice). |
| Predominantly nuclear staining with little cytoplasmic signal. | An exclusively nuclear pattern conflicts with the reported cytoplasmic tissue pattern and cytoplasm and lysosome annotations (HPA tissue IHC: ubiquitous cytoplasmic expression; UniProt P30041: subcellular location). Review background, antibody specificity, and cell boundaries before scoring it as PRDX6 (standard IHC practice). |
| Strong staining in a cell population expected to stain weakly, such as soft-tissue fibroblasts. | Fibroblasts are reported at a low level, so unexpectedly strong signal warrants checking cell identity and controls for cross-reactivity or endogenous detection activity (HPA tissue IHC: Low in soft-tissue fibroblasts; standard IHC practice). Low does not mean absent (HPA tissue IHC: Low). |
| Diffuse color over cells and surrounding tissue, without a discernible cellular pattern. | Nonlocalized haze cannot establish PRDX6 expression; compare a no-primary control and assess blocking, washing, and detection conditions (standard IHC practice). The expected tissue pattern is cytoplasmic within cells (HPA tissue IHC: ubiquitous cytoplasmic expression). |
| No staining in adipocytes, bronchial respiratory epithelial cells, or marrow hematopoietic cells. | These are reported high-staining reference cells, so absent signal first calls for an assay check (HPA tissue IHC: High). Confirm that the expected cells are present, then review antibody dilution, retrieval, and detection controls (standard IHC practice). A blank section alone cannot establish biological absence. |
💡Expected PRDX6 appearanceCall a result positive when identifiable cells show cytoplasmic staining, potentially strong in HPA high-staining cell types; isolated nuclear color or diffuse background is suspect (HPA tissue IHC: ubiquitous cytoplasmic expression; High in adipocytes and bronchial respiratory epithelial cells; standard IHC practice).
| Tissue and cell context | PRDX6 has low tissue specificity, with high and low cell-level examples (HPA tissue IHC: RNA low tissue specificity; High in adipocytes; Low in urothelial cells). Compare the same cell type when judging intensity; a low example is a contrast, not a negative control (standard IHC practice). |
| Compartment and topology | Cytoplasm and lysosome are annotated, with no transmembrane segment; lung lamellar bodies are also noted (UniProt P30041: subcellular location; topology). Interpret discrete intracellular signal in its tissue context, without requiring every positive cell to show visible puncta. |
| Evidence boundary between IHC and IF/ICC | Tissue IHC reports ubiquitous cytoplasmic expression, while ICC-IF places PRDX6 mainly at the plasma membrane and cytosol (HPA tissue IHC: profile; HPA subcellular ICC-IF: approved membrane, supported cytosol). Apply each observation to its own preparation; do not require a membrane rim in chromogenic sections. |
| Antibody evidence | Two listed antibodies have Supported IHC status; one also has Supported ICC status (HPA antibodies: HPA006983 and CAB008663). This supports using the tissue pattern as a reference, while the tissue profile's medium RNA–staining consistency limits certainty for any single cell (HPA tissue IHC: reliability description). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining cells remain blank. | The result disagrees with reported high staining; antibody dilution, retrieval, or detection may need review (HPA tissue IHC: High in adipocytes and bronchial respiratory epithelial cells; standard IHC practice). | Verify the cells on the section and inspect the run's positive and detection controls; adjust one assay condition at a time (standard IHC practice). Do not score the tissue negative from this run alone. |
| Color covers tissue and empty spaces. | Diffuse deposition or incomplete blocking can obscure the expected cytoplasmic pattern (standard IHC practice; HPA tissue IHC: ubiquitous cytoplasmic expression). | Compare a no-primary control, then review blocking, washing, and detection exposure (standard IHC practice). Score only signal that can be assigned to cells. |
| Signal is mainly nuclear. | The compartment disagrees with cytoplasmic tissue staining and the annotated cytoplasm and lysosome locations (HPA tissue IHC: profile; UniProt P30041: subcellular location). | Check the counterstain and cell boundaries, then compare controls and the antibody's validation evidence before attributing nuclear color to PRDX6 (standard IHC practice). |
| Fibroblasts or urothelial cells stain more strongly than the chosen high-staining reference cells. | That ranking differs from the supplied tissue observations, though staining can vary between specimens (HPA tissue IHC: Low in soft-tissue fibroblasts and urothelial cells; High in adipocytes). | Recheck cell identity and compare like tissue regions; examine no-primary and detection controls for nonspecific or endogenous color (standard IHC practice). Do not treat a low HPA level as zero. |
| A weak cell population looks negative at first glance. | Low reported staining may be difficult to separate from counterstain or background (HPA tissue IHC: Low in esophageal squamous epithelial cells; standard IHC practice). | Use the same scoring threshold across comparable cells and check the positive reference and background controls (standard IHC practice). Report weak or indeterminate staining without converting it into proven absence. |
| IF/ICC Q: Is a plasma-membrane rim compatible with PRDX6? | The ICC-IF summary reports a mainly plasma-membrane and cytosol pattern (HPA subcellular ICC-IF: membrane approved; cytosol supported). | Yes, interpret a rim together with cytosolic signal in IF/ICC (HPA subcellular ICC-IF: main locations). Use the separate IF/ICC guide for that assay; the chromogenic IHC tissue reference remains cytoplasmic (HPA tissue IHC: profile). |
Experiment evidence
IHC & IF images showing PRDX6 staining
PB9350 · IHCIHC analysis of PRDX6 using anti-PRDX6 antibody (PB9350). PRDX6 was detected in paraffin-embedded section of Human Mammary Cancer tissues. H
PB9350 · IHCIHC analysis of PRDX6 using anti-PRDX6 antibody (PB9350). PRDX6 was detected in paraffin-embedded section of human gastric cancer tissues. H
PB9350 · IFICC analysis of PRDX6 using anti-PRDX6 antibody (PB9350). PRDX6 was detected in immunocytochemical section of PC-3 cell. Enzyme antigen retr
M01847-1 · IHCIHC analysis of PRDX6 using anti-PRDX6 antibody (M01847-1). PRDX6 was detected in paraffin-embedded section of human endometrial carcinoma t
M01847-1 · IFIF analysis of PRDX6 using anti-PRDX6 antibody (M01847-1). PRDX6 was detected in immunocytochemical section of A549. Enzyme antigen retrieva
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117592-PRDX6/tissue/cerebral+cortex#imid_2126152
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117592-PRDX6/tissue/colon#imid_2126062
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117592-PRDX6/tissue/liver#imid_2126071
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000117592-PRDX6/tissue/kidney#imid_2126080
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB008663; green = PRDX6, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB008663; green = PRDX6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRDX6 IHC & IF
🧪Run adipose tissue first: adipocytes should stain (HPA: High in adipocytes). HPA detects PRDX6 in all 45 scored tissues and lists no negative tissue (HPA: negative rows empty); use no-primary and isotype controls for the negative comparison, and expect cells without specific staining on the positive slide to show counterstain alone rather than treating any cell type as a proven internal negative.
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: PRDX6 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRDX6 in A-431, U-251MG, U2OS, with annotated localisation: Plasma membrane (approved), Cytosol (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and a rabbit IgG isotype control matched to the primary antibody’s clonality (selected-SKU caption: rabbit primary); a PRDX6 knockout sample, if available, provides a biological negative. For chromogenic IHC, block endogenous peroxidase and check for endogenous biotin signal with the caption’s biotin-based detection system (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). That caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required; in adipose tissue, assess staining in the thin cytoplasmic rim around lipid vacuoles (selected-SKU caption: retrieval conditions; HPA: High in adipocytes). IF/ICC: is it easier than paraffin IHC? HPA has ICC-IF images from A-431, U-251MG and U2OS, but the supplied evidence does not establish whether IF/ICC or frozen sections are easier for PRDX6 (HPA: ICC-IF image cell lines).
HPA tissue IHC evidence for PRDX6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PRDX6 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced PRDX6 IHC Tips
Troubleshoot PRDX6 staining in paraffin sections by checking retrieval, compartment, controls, and scoring before interpreting changes in signal.
Which retrieval condition should I try first for PRDX6 in paraffin sections?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet PB9350; selected PB9350 tissue-IHC caption). Use a comparable tissue section and the same antibody concentration when assessing whether retrieval improved staining (standard IHC practice). The selected image used 1 µg/mL antibody overnight at 4°C, giving a documented starting point for that comparison (selected PB9350 tissue-IHC caption). If staining remains weak, assess a different retrieval buffer or heating time on adjacent sections while monitoring tissue damage (standard IHC practice). Interpret gains against expected cytoplasmic staining, since PRDX6 has a cytoplasmic location (UniProt P30041; HPA tissue IHC).
Could fixation explain weak or uneven PRDX6 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (selected PB9350 tissue-IHC caption). Record each specimen’s fixative, fixation duration, and processing history before comparing staining across blocks (standard IHC practice). On matched sections, keep citrate retrieval at pH 6 for 20 minutes and the primary antibody at 1 µg/mL while investigating variation (selected PB9350 tissue-IHC caption). Check whether weak staining tracks section-wide processing differences or only particular cells, and include a tissue with a documented PRDX6 staining pattern (standard IHC practice; HPA tissue IHC). Do not infer a fixation effect from PRDX6 localization or modified residues (UniProt P30041).
Should PRDX6 staining appear cytoplasmic, membranous, or punctate?
Expect broad cytoplasmic staining in tissue IHC, consistent with the reported ubiquitous cytoplasmic profile (HPA tissue IHC). A peripheral component can be biologically plausible because plasma membrane and cytosol are reported cellular locations (HPA subcellular). Puncta may warrant closer examination because lysosomes and lung lamellar bodies are also listed locations (UniProt P30041). PRDX6 has no transmembrane segment, so a sharp, exclusive membrane outline should prompt a review of morphology and controls (UniProt P30041 topology; standard IHC practice). Compare the pattern across adjacent cells and sections before assigning a compartment from DAB alone, whose deposits have limited spatial resolution (standard IHC practice).
How should epitope uncertainty affect PRDX6 IHC troubleshooting?
The supplied record lists 0 isoforms and a thioredoxin domain at residues 5–169, but it does not map the catalog antibody’s epitope (UniProt P30041; supplied antibody evidence). PRDX6 has no transmembrane segment or annotated glycosylation sites, which does not establish epitope accessibility after tissue processing (UniProt P30041 topology and glycosylation). Reported modifications include residues 44, 63, 89, 177, and 209; their effect on this antibody’s staining is unknown (UniProt P30041 modified residues). If localization or intensity is unexpected, compare independently validated antibodies or appropriate biological controls before attributing the result to a particular modification (standard IHC practice).
How can IF help resolve an ambiguous PRDX6 IHC pattern?
Use the separate IF/ICC guide to plan fluorescence staining; this page’s selected antibody procedure describes paraffin-section chromogenic IHC (selected PB9350 tissue-IHC caption). For multiplexing, pair PRDX6 with a marker for the expected cell population, such as a respiratory epithelial marker when examining bronchus, where respiratory epithelial cells stain highly (HPA tissue IHC; standard IF practice). Choose fluorophores whose emission can be distinguished from tissue autofluorescence, and include single-color controls (standard IF practice). Because PRDX6 has no transmembrane segment and has cytosolic and lysosomal locations, assess permeabilisation for intracellular access rather than assuming surface-only staining (UniProt P30041 topology and location; HPA subcellular).
How do I distinguish PRDX6 signal from chromogenic background?
Run a no-primary control and inspect it for endogenous enzyme activity and nonspecific detection deposits before judging faint DAB staining (standard IHC practice). Include a peroxidase block in the chromogenic workflow and assess whether the block reduces background on a matched section (standard IHC practice). The selected procedure used 10% goat serum, biotinylated secondary antibody, a streptavidin–biotin complex, and DAB, so evaluate background from those steps separately (selected PB9350 tissue-IHC caption). PRDX6 staining is reported as broadly cytoplasmic, making a completely unstained tissue region a poor universal negative control (HPA tissue IHC). Compare signal with tissue morphology and the no-primary control at matched imaging settings (standard IHC practice).
What is a defensible way to quantify PRDX6 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC reports widespread cytoplasmic PRDX6 expression (HPA tissue IHC). For DAB sections, record the percentage of positive cells and staining intensity, then calculate an H-score using 0–3 intensity grades if that scale is prespecified (standard IHC practice). Where cell counts are unreliable, measure positive area or cell density per mm² within consistently defined regions (standard IHC practice). Normalize comparisons to the relevant cell population or analyzable tissue area, rather than raw whole-section darkness (standard IHC practice). Keep retrieval, detection, exposure, and threshold settings consistent across specimens (standard IHC practice).
When should an apparent PRDX6-positive region be treated cautiously?
A credible signal should fit cell morphology and the mainly cytoplasmic tissue pattern, while allowing a possible peripheral component (HPA tissue IHC; HPA subcellular). Review exclusive nuclear staining or a crisp surface-only outline carefully, given the reported cytosolic location and absence of a transmembrane segment (HPA subcellular; UniProt P30041 topology). Strong staining at section edges, necrotic regions, or areas positive in the no-primary control can indicate artefact or endogenous enzyme signal (standard IHC practice). Compare suspect cells with nearby intact cells and a documented positive population, such as bronchial respiratory epithelium (HPA tissue IHC; standard IHC practice). A single DAB pattern does not establish PRDX6 enzymatic activity (UniProt P30041 function; standard IHC practice).
Boster reagents
Best PRDX6 / Peroxiredoxin-6 IHC Antibodies
Human paraffin tissue IHC and A549 cell IF are pictured; catalog reactivity includes human, mouse, and rat (IHC/IF image captions; catalog reactivity).
Real IHC data
Anti-Peroxiredoxin 6/PRDX6 Antibody ®
Cat # PB9350
Real IHC data
Anti-PRDX6 Antibody ® (monoclonal, 6I8)
Cat # M01847-1
PB9350 shows PRDX6 IHC in human mammary, gastric, lung, and ovarian cancer paraffin sections (PB9350 IHC captions). M01847-1 shows human endometrial carcinoma paraffin IHC and A549 cell IF (M01847-1 IHC/IF captions).
Which to pick: For human paraffin tissue IHC, PB9350 uses citrate retrieval at pH 6 for 20 minutes, while M01847-1 uses EDTA retrieval at pH 8; neither IHC caption reports the fixative (respective IHC captions). For cell IF/ICC, choose mouse monoclonal M01847-1: its A549 IF image uses 2 μg/ml, while PB9350’s cell image uses chromogenic detection (M01847-1 title/IF caption; PB9350 ICC caption). For cross-species planning, PB9350 lists human, mouse, and rat reactivity, but its IHC dilution is specified for human and rat; M01847-1 lists human and rat reactivity, with an IHC dilution specified for human (catalog reactivity/dilution).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P30041 (PRDX6_HUMAN, Peroxiredoxin-6).
- Human Protein Atlas. PRDX6 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. PRDX6 subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol..
- Human Protein Atlas. PRDX6 antibody validation summary (2 antibodies).
- PRDX6 knockout restrains the malignant progression of intrahepatic cholangiocarcinoma. Medical oncology (Northwood, London, England) 2022 — PMC9547796.
- Overexpression of Peroxiredoxin 6 (PRDX6) Promotes the Aggressive Phenotypes of Esophageal Squamous Cell Carcinoma. Journal of Cancer 2018 — PMC6218759.
- Effects of buyang huanwu decoction on ventricular remodeling and differential protein profile in a rat model of myocardial infarction. Evidence-based complementary and alternative medicine : eCAM 2012 — PMC3459299.
- Quantitative HDL Proteomics Identifies Peroxiredoxin-6 as a Biomarker of Human Abdominal Aortic Aneurysm. Scientific reports 2016 — PMC5146935.
- PubMed PMID:8999971 — UniProt-cited evidence.
- PubMed PMID:9050990 — UniProt-cited evidence.
- PubMed PMID:7788527 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRDX6 (Peroxiredoxin-6). Expected localisation Cytoplasmic staining in tissue sections (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet PB9350). Validated IHC antibody: PB9350 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human Mammary Cancer tissues; fixative not specified (datasheet PB9350); 2. Fixation: Image fixative and duration unreported (datasheet PB9350); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6 , 20 min (datasheet PB9350); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9350); 8. Primary antibody: Rabbit anti-PRDX6, 0.5-1μg/ml (datasheet PB9350); 9. Primary incubation: Overnight at 4 °C (datasheet PB9350). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); optimize locally.; Caveat: Urothelial cells show low staining (HPA tissue IHC); Isoform / epitope: No isoforms annotated; chain spans residues 2–224 (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt P30041, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol., Datasheet PB9350, PMC3459299, PMC5146935. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRDX6/immunohistochemistry-ihc-guide.