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PRDX1 / Peroxiredoxin-1 · IHC design guide
Design Immunohistochemistry for PRDX1
Plan PRDX1 chromogenic IHC on paraffin sections with a catalog antibody tested at 1 μg/ml (datasheet PB9348). Use the tissue IHC profile to assess cytoplasmic staining and select high-staining bone marrow, spleen or thyroid compartments as positive references (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PRDX1 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining across most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9348) | |
| Positive control | Bone marrow+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Endogenous peroxidase in marrow may mimic DAB signal (HPA tissue IHC; standard IHC practice) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms; map epitopes to residues 2–199 (UniProt) |
Section 1
Recommended PRDX1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet PB9348). The options below summarize three published PRDX1 IHC methods (PMC9353087; PMC5777129; PMC4327392).
| Sample | Paraffin-embedded Human Mammary Cancer tissue; fixative not specified (datasheet PB9348) |
| Fixation | Image fixative and duration unreported (datasheet PB9348); 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 PB9348); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9348) |
| Primary antibody | Rabbit anti-PRDX1, 0.5-1μg/ml (datasheet PB9348) |
| Primary incubation | Overnight at 4 °C (datasheet PB9348) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PB9348) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PRDX1-positive staining in hematopoietic cells of bone marrow (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 retrieval for the catalog antibody (datasheet PB9348); the canine study used a pH 6.0 solution with a different antibody (PMC9353087).
Section 2
What Is the Expected PRDX1 Staining Pattern?
In paraffin-section IHC, expect predominantly cytoplasmic PRDX1 staining across many tissues, with strong examples in bone-marrow hematopoietic cells, splenic red-pulp cells, and thyroid glandular cells (HPA tissue IHC: High; cytoplasmic expression in most tissues). PRDX1 has no transmembrane segment (UniProt Q06830 topology). HPA rates the tissue pattern Approved, with medium agreement between staining and RNA data and external verification pending (HPA tissue IHC).
| Cytoplasmic staining is strong in bone-marrow hematopoietic cells or thyroid glandular cells. | This matches the expected compartment and two High tissue examples (HPA tissue IHC). Score the relevant cells, their staining intensity, and background separately; an HPA High designation describes an observed pattern, not a required intensity in every section. |
| Staining is predominantly nuclear, with little cytoplasmic signal in otherwise positive cells. | Treat this as a compartment mismatch and investigate nonspecific staining or detection artefact before scoring it as PRDX1. The tissue IHC profile is cytoplasmic (HPA tissue IHC); UniProt lists cytoplasm and melanosome, while HPA separately reports mitochondrial localisation by ICC-IF (UniProt Q06830; HPA subcellular ICC-IF). |
| Adipocytes or cardiomyocytes stain as strongly as the expected positive cells. | These cell types are listed as Not detected in HPA tissue IHC. Check for cross-reactivity or endogenous detection activity, especially if the signal also appears in a no-primary control (HPA tissue IHC; general IHC practice). A single discrepant section does not establish a new tissue pattern. |
| Diffuse colour covers stroma, lumens, or the entire section without clear cell boundaries. | This does not show the cell-associated cytoplasmic distribution expected from the HPA tissue profile. Examine the no-primary control, blocking, washes, and detection system for general background (HPA tissue IHC; general IHC practice). Do not score diffuse deposit as positive cells. |
| There is no cytoplasmic signal in bone-marrow hematopoietic cells or splenic red-pulp cells. | These are High examples, so an absent signal calls for a run-level check before a biological interpretation (HPA tissue IHC). Confirm tissue identity, antibody suitability for IHC-P, retrieval and detection steps, then compare with a processed positive control (general IHC practice). |
💡Expected PRDX1 appearanceCall a convincing positive result when identifiable hematopoietic, splenic red-pulp, or thyroid glandular cells show cell-associated cytoplasmic staining, potentially strong in these High examples; broad acellular colour or isolated nuclear staining is suspect (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | Use named High cell populations as positive comparators and the named Not detected cell populations as background checks (HPA tissue IHC). HPA reports low tissue RNA specificity and cytoplasmic expression in most tissues, so a whole tissue should not be treated as uniformly positive or negative (HPA tissue IHC). |
| Compartment evidence and the IF/ICC question | Q: Should mitochondrial IF/ICC signal define the IHC result? A: No direct equivalence is established. HPA calls mitochondria the approved main ICC-IF location, whereas its tissue IHC profile is cytoplasmic; UniProt also lists cytoplasm and melanosome, with melanosome identification from fractions (HPA subcellular ICC-IF; HPA tissue IHC; UniProt Q06830). |
| Antibody evidence | Two listed antibodies have Approved IHC status, while the overall tissue profile has medium staining-to-RNA consistency and awaits external verification (HPA antibodies; HPA tissue IHC). Interpret an unexpected pattern cautiously and compare controls; Approved does not make every compartment or cell-type signal specific. |
| Protein architecture and preparation limits | PRDX1 has no transmembrane segment or signal peptide, and the listed chain starts at residue 2 (UniProt Q06830 topology and processing). These annotations give no basis for expecting a membrane rim or shed extracellular stain. The supplied UniProt and HPA records give no target-specific fixation sensitivity or retrieval requirement. |
| Situation | Likely cause | Next action |
|---|---|---|
| A High example shows no staining. | The run may have failed, or the selected detection conditions may be unsuitable; HPA documents High staining in bone-marrow hematopoietic and splenic red-pulp cells (HPA tissue IHC). | Check tissue identity and a processed positive control. Review the IHC-P antibody instructions, retrieval, primary incubation, and detection steps (general IHC practice). |
| Signal is nuclear with little cytoplasmic staining. | A compartment mismatch may reflect nonspecific staining or a detection artefact; the tissue IHC pattern is cytoplasmic (HPA tissue IHC). | Compare with a no-primary control and a named positive cell population. Reassess localisation before scoring the slide (general IHC practice). |
| Adipocytes or cardiomyocytes appear strongly positive. | These are Not detected examples in the HPA tissue IHC record; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Check a no-primary control. If using peroxidase-based detection, review endogenous peroxidase blocking and whether deposit follows identifiable cells (general IHC practice). |
| Colour is diffuse across the section. | General background from detection, blocking, or washing can obscure cell-associated staining (general IHC practice). | Inspect the no-primary control and wash and blocking steps; score only interpretable cell-associated cytoplasmic signal (HPA tissue IHC; general IHC practice). |
| A negative comparator has faint staining. | Not detected is an observed HPA category, not proof that every cell or section must be absolutely colourless (HPA tissue IHC). | Compare the faint signal with local background and the High comparator on the same run; record cell type and intensity before calling it positive (general IHC practice). |
| Mitochondrial IF/ICC signal seems inconsistent with IHC. | HPA reports an approved mitochondrial ICC-IF location and a cytoplasmic tissue IHC profile using different assays (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret each assay against its own control and compartment evidence. Use the separate IF/ICC guide for fluorescence workflow; do not infer an IHC-P retrieval setting from the ICC-IF image. |
Experiment evidence
IHC & IF images showing PRDX1 staining
PB9348 · IHCIHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348). Peroxiredoxin 1 was detected in a paraffin-embedded section of
PB9348 · IHCIHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348). Peroxiredoxin 1 was detected in a paraffin-embedded section of
PB9348 · IFIF analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348). Peroxiredoxin 1 was detected in immunocytochemical section of H
M01845-3 · IHCRat kidney was stained with anti-Peroxiredoxin 1 rabbit antibody
M01845-3 · IHCHuman kidney was stained with anti-Peroxiredoxin 1 rabbit antibody
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000117450-PRDX1/tissue/cerebral+cortex#imid_1367148
HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000117450-PRDX1/tissue/thyroid+gland#imid_1367108
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000117450-PRDX1/tissue/colon#imid_1367090
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000117450-PRDX1/tissue/liver#imid_1367083
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007730; green = PRDX1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007730; green = PRDX1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PRDX1 IHC & IF
🧪Run thyroid gland first: its glandular cells should stain (HPA: High in thyroid gland glandular cells). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the thyroid slide, cells without target signal should show counterstain and background only, but the supplied HPA rows do not establish a specific thyroid cell type as PRDX1-negative.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PRDX1 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and matched rabbit IgG controls, using normal rabbit IgG for a polyclonal primary or the matching rabbit IgG subclass for a monoclonal primary; validate specificity with a PRDX1-knockout specimen where available (PB9348 caption: rabbit primary; standard IHC practice). Quench endogenous peroxidase before HRP/DAB detection in thyroid tissue and check the no-primary slide for residual chromogen (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected PB9348 paraffin-section caption does not state a fixative (PB9348 caption). That caption uses heat-mediated EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody overnight at 4°C, but does not establish that retrieval is required (PB9348 caption). ICC-IF images are available for A-431, U-251MG and U2OS (HPA: subcellular data), but the supplied evidence does not establish whether IF or frozen sections are easier; endogenous thyroid peroxidase can produce misleading HRP/DAB signal (standard IHC practice).
HPA tissue IHC evidence for PRDX1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | 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 → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
| Vagina | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PRDX1 IHC Tips
Troubleshoot PRDX1 staining in paraffin sections by checking retrieval, controls, cellular localisation and scoring before interpreting differences in signal.
Which retrieval condition should I try first for PRDX1 IHC?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet PB9348). The selected tissue-IHC image used that condition before incubation with 1 μg/ml primary antibody overnight at 4°C (datasheet PB9348). If staining is weak, compare a small retrieval time series on adjacent sections while keeping antibody concentration, detection and imaging conditions fixed (standard IHC practice). Use a consistently stained control, such as thyroid glandular cells, to judge whether retrieval improved cellular signal (HPA: High in thyroid glandular cells). Record tissue damage and background alongside signal, since stronger staining alone does not establish specificity (standard IHC practice).
Could fixation explain weak or uneven PRDX1 staining?
The selected paraffin-section caption does not state its fixative, so PRDX1-specific fixation sensitivity is unknown (datasheet PB9348). Do not infer an optimal fixation duration from PRDX1 topology or modification sites; those annotations do not test preservation of its IHC epitope (UniProt Q06830). Compare sections with documented processing histories using the same EDTA pH 8.0 retrieval and detection settings (datasheet PB9348; standard IHC practice). Check whether weak staining tracks processing batches, section folds or damaged areas rather than a defined cell population (standard IHC practice). Interpret a batch effect as a processing concern requiring controlled comparison, not as evidence of altered PRDX1 abundance (standard IHC practice).
How should I assess cytoplasmic versus mitochondrial PRDX1 staining?
Score the dominant cellular pattern first: tissue IHC reports cytoplasmic expression in most tissues (HPA: tissue IHC), while UniProt lists cytoplasm and melanosome (UniProt Q06830). A separate subcellular IF assessment assigns an approved mitochondrial location (HPA: subcellular), so punctate signal merits comparison with a mitochondrial marker in a separate assay. PRDX1 has no transmembrane segment; crisp cell-surface-only staining would therefore need independent validation (UniProt Q06830 topology). Examine a consistently stained control and a no-primary section under the same chromogenic conditions (standard IHC practice). Describe cytoplasmic, punctate and nuclear signal separately rather than treating every brown deposit as equivalent PRDX1 staining (standard IHC practice).
Can isoforms or epitope masking account for inconsistent staining?
UniProt lists 0 PRDX1 isoforms in this record, so an annotated isoform switch does not explain a change in staining (UniProt Q06830). It places the thioredoxin domain at residues 6–165 and lists modified residues, but the supplied evidence does not map this antibody’s epitope (UniProt Q06830; datasheet PB9348). Oxidation can form disulfide-linked PRDX1 homodimers, yet an effect on this antibody’s staining has not been established here (UniProt Q06830). Keep retrieval, primary incubation and detection matched when comparing specimens (standard IHC practice). If staining differs unexpectedly, seek an independently validated antibody with a documented epitope before assigning the difference to PRDX1 modification (standard IHC practice).
How can IF help investigate an ambiguous PRDX1 IHC pattern?
Use IF as a separate localisation check, since subcellular IF reports approved mitochondrial localisation whereas tissue IHC reports cytoplasmic expression in most tissues (HPA: subcellular; HPA: tissue IHC). Multiplex PRDX1 with a marker identifying the expected cell population, such as glandular cells in thyroid, and compare cellular overlap rather than brightness alone (HPA: High in thyroid glandular cells; standard IF practice). Select fluorophores in channels with low measured tissue autofluorescence and include single-label controls for bleed-through (standard IF practice). Because PRDX1 has no transmembrane segment and is annotated in the cytoplasm, optimise permeabilisation for access to intracellular epitopes (UniProt Q06830 topology and localisation; standard IF practice). Do not transfer the paraffin-section antibody conditions directly to IF without validating that assay (datasheet PB9348; standard IF practice).
What should I check when PRDX1 DAB staining looks diffuse?
The selected paraffin-section example used 10% goat serum blocking, 1 μg/ml primary antibody and DAB detection (datasheet PB9348). Run a no-primary control, check endogenous peroxidase blocking and compare background after adequate washes; these are general chromogenic IHC checks (standard IHC practice). PRDX1 is reported as cytoplasmic in most tissues, so diffuse colour outside cells is not convincing localisation by itself (HPA: tissue IHC). Examine section edges, folds and damaged regions separately from intact tissue before changing antibody concentration (standard IHC practice). If background persists, optimise one variable at a time against a control tissue with expected staining (HPA: High in thyroid glandular cells; standard IHC practice).
How should I quantify PRDX1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC reports cytoplasmic expression across many tissues (HPA: tissue IHC). For cellular comparisons, record the percentage of positive cells and staining intensity, then calculate an H-score using a fixed scoring rubric (standard IHC practice). For spatial questions, report positive-cell density per mm² of evaluable tissue and specify excluded necrotic or damaged regions (standard IHC practice). Normalise counts to the number of eligible cells or evaluable area, and keep retrieval, DAB development and image settings consistent across groups (standard IHC practice). Stratify different cell populations rather than pooling them when their expected staining differs (HPA: tissue IHC).
When is PRDX1 staining convincing rather than artefactual?
A convincing pattern places signal within intact cells and matches a plausible compartment and cell population: tissue IHC describes predominantly cytoplasmic staining, including high staining in bone marrow hematopoietic cells (HPA: tissue IHC). Treat isolated surface-only or nuclear-only staining cautiously because UniProt annotates cytoplasm and no transmembrane segment (UniProt Q06830 localisation and topology). Compare no-primary controls to identify detection artefacts, including residual endogenous peroxidase activity (standard chromogenic IHC practice). Exclude edge staining, folds and necrotic regions from interpretation and scoring (standard IHC practice). HPA tissue-IHC reliability is Approved with medium staining-to-RNA consistency and pending external verification, so corroborate unexpected biological claims independently (HPA: tissue IHC reliability).
Boster reagents
Best PRDX1 / Peroxiredoxin-1 IHC Antibodies
Anti-PRDX1 IHC images cover human mammary cancer, kidney and thyroid, mouse and rat brain, and rat kidney (catalog image captions); IF/ICC images show HeLa and U20S cells (PB9348 IF captions).
Real IHC data
Anti-Peroxiredoxin 1/PRDX1 Antibody ®
Cat # PB9348
Real IHC data
Anti-Peroxiredoxin 1 Rabbit Monoclonal Antibody
Cat # M01845-3
PB9348 has paraffin-section IHC images from human mammary cancer and mouse and rat brain, plus IF/ICC images from HeLa and U20S cells (PB9348 image captions). M01845-3 has IHC images from rat and human kidney and human thyroid; IF is also a listed application (M01845-3 image captions; catalog applications).
Which to pick: For paraffin-section tissue IHC, PB9348 has a caption documenting EDTA retrieval at pH 8.0 and 1 μg/ml primary antibody; the fixative is unreported (PB9348 IHC caption). For IF/ICC, choose PB9348 if a cell-image example is useful; M01845-3 lists IF but has no IF image in the payload (PB9348 IF captions; M01845-3 catalog applications and image alts). For work across human, mouse and rat samples, both list all three species; PB9348 shows mouse and rat brain IHC, while M01845-3 shows rat and human kidney IHC (catalog reactivity; respective IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q06830 (PRDX1_HUMAN, Peroxiredoxin-1).
- Human Protein Atlas. PRDX1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PRDX1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. PRDX1 antibody validation summary (2 antibodies).
- The immunohistochemical detection of peroxiredoxin 1 and 2 in canine spontaneous vascular endothelial tumors. The Journal of veterinary medical science 2022 — PMC9353087.
- Peroxiredoxin 1 expression in active ulcerative colitis mucosa identified by proteome analysis and involvement of thioredoxin based on immunohistochemistry. Oncology letters 2018 — PMC5777129.
- Peroxiredoxin 1 promotes pancreatic cancer cell invasion by modulating p38 MAPK activity. Pancreas 2015 — PMC4327392.
- PRDX1 is a Tumor Suppressor for Nasopharyngeal Carcinoma by Inhibiting PI3K/AKT/TRAF1 Signaling. OncoTargets and therapy 2020 — PMC7501964.
- PubMed PMID:8496166 — UniProt-cited evidence.
- PubMed PMID:8026862 — UniProt-cited evidence.
- PubMed PMID:19054851 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PRDX1 (Peroxiredoxin-1). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9348). Validated IHC antibody: PB9348 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human Mammary Cancer tissue; fixative not specified (datasheet PB9348); 2. Fixation: Image fixative and duration unreported (datasheet PB9348); 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 PB9348); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9348); 8. Primary antibody: Rabbit anti-PRDX1, 0.5-1μg/ml (datasheet PB9348); 9. Primary incubation: Overnight at 4 °C (datasheet PB9348). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Endogenous peroxidase in marrow may mimic DAB signal (HPA tissue IHC; standard IHC practice); Isoform / epitope: No annotated isoforms; map epitopes to residues 2–199 (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q06830, HPA tissue IHC · Approved, HPA ICC-IF · mitochondria., Datasheet PB9348, PMC9353087, PMC5777129. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PRDX1/immunohistochemistry-ihc-guide.