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- Table of Contents
Plan PRDX1 staining in paraffin sections using cytoplasmic expression in most tissues as the expected pattern (HPA tissue IHC). This guide covers fixation consistency, antibody selection and chromogenic staining controls.
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in most tissues (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 |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet PB9348); verify before use. | |
| Caveat | Endogenous peroxidase may raise marrow background (HPA tissue IHC; standard IHC practice) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans residues 2–199 (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet: PB9348) is accompanied by 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. |
In paraffin-section IHC, expect predominantly cytoplasmic PRDX1 staining across many tissues, including high staining in bone-marrow hematopoietic cells, splenic red-pulp cells, and thyroid glandular cells (HPA: cytoplasmic in most tissues; High in these cells). PRDX1 has no transmembrane segment or signal peptide (UniProt Q06830 topology and processing). Treat this as an expected pattern, not a universal positive result: HPA rates its tissue-IHC evidence Approved, with medium consistency and pending external verification (HPA: reliability).
| Cytoplasmic staining in bone-marrow hematopoietic cells, splenic red-pulp cells, or thyroid glandular cells (HPA: High in these cells). | This matches the reported compartment and high-staining examples (HPA: tissue IHC). Compare stained cells with their neighbors; HPA's High label describes those cell populations, not every cell in the section (HPA: tissue IHC). |
| Predominantly nuclear staining, or staining confined to cell borders, with little cytoplasmic signal. | The compartment differs from the reported tissue-IHC pattern (HPA: cytoplasmic in most tissues). Check morphology, counterstain, and control staining before calling it PRDX1; compartment mismatch raises concern for artefact or nonspecific detection (general IHC practice). |
| Strong staining in adipocytes, cardiomyocytes, soft-tissue fibroblasts, or vaginal squamous cells (HPA: Not detected in these cells). | These are reported negative cell populations, so strong staining there warrants a cross-reactivity or endogenous-detection check (HPA: tissue IHC; general IHC practice). A negative call applies to the named cells, not necessarily every other cell in those tissues (HPA: tissue IHC). |
| Color spreads evenly over cells and surrounding section, obscuring cytoplasmic boundaries. | A diffuse, non-cell-specific deposit does not reproduce HPA's cytoplasmic pattern (HPA: tissue IHC). Compare a no-primary control and inspect blocking, washing, and detection chemistry as general IHC troubleshooting steps (general IHC practice). |
| No staining in the expected cells of an otherwise interpretable positive-control section. | Absence in bone-marrow hematopoietic cells, splenic red-pulp cells, or thyroid glandular cells conflicts with HPA's High examples (HPA: tissue IHC). Check antibody, retrieval, detection, and control performance before treating the specimen as biologically negative (general IHC practice). |
| Which cells provide the clearest tissue-IHC comparison? | HPA reports High staining in bone-marrow hematopoietic cells, splenic red-pulp cells, and thyroid glandular cells; Medium staining in appendix and breast glandular cells, bronchial respiratory epithelium, caudate glia, and cerebellar granular-layer cells (HPA: tissue IHC). Score the specified cells and compartment, because tissue-wide intensity can obscure cell-level disagreement (general IHC practice). |
| How firm is the reference pattern? | The tissue-IHC profile is Approved, with medium agreement between antibody staining and RNA expression and external verification pending (HPA: reliability). HPA lists one rabbit polyclonal and one mouse monoclonal antibody as IHC Approved; that status alone does not establish an Enhanced validation designation (HPA: antibody validation). |
| Does PRDX1 topology support a surface pattern? | UniProt places PRDX1 in the cytoplasm and reports no signal peptide or transmembrane segment (UniProt Q06830: subcellular location, topology, processing). Those annotations support an intracellular expectation; they do not identify the antibody epitope or establish any target-specific retrieval or fixation sensitivity (UniProt Q06830: topology and processing). |
| Does protein processing predict a separate staining compartment? | UniProt reports no propeptide, a chain spanning residues 2–199, and no annotated isoforms (UniProt Q06830: processing and isoforms). Its melanosome annotation comes from mass spectrometry of melanosome fractions; that observation alone does not predict a distinct chromogenic tissue-IHC pattern (UniProt Q06830: subcellular location). |
| IF/ICC Q&A: Should mitochondrial fluorescence change the IHC call? | HPA's ICC-IF summary calls the main location mitochondria, while its tissue-IHC profile says cytoplasmic expression in most tissues (HPA: subcellular ICC-IF; HPA: tissue IHC). Interpret the paraffin-section chromogenic result against the tissue-IHC profile; record the assay difference without assuming either observation is an error (HPA: assay-specific observations). |
| Situation | Likely cause | Next action |
|---|---|---|
| A high-expression reference section shows little or no cytoplasmic stain. | A failed staining run is possible; HPA reports High staining in the named reference cells, but does not supply a PRDX1-specific fixation-sensitivity claim (HPA: tissue IHC). | Confirm the correct cells are present, then check the IHC-validated antibody's stated IHC-P conditions, retrieval, dilution, and detection controls (general IHC practice). |
| The whole section is dark, including areas without plausible cellular signal. | Diffuse deposit can reflect background from detection chemistry or incomplete blocking or washing (general IHC practice); it is unlike the reported cytoplasmic pattern (HPA: tissue IHC). | Review a no-primary control, blocking and wash steps, and chromogen development; assess cell boundaries again after background is reduced (general IHC practice). |
| Only nuclei or sharply outlined cell borders are prominent. | This compartment conflicts with HPA's cytoplasmic tissue-IHC profile (HPA: tissue IHC). It does not, by itself, identify the source of the mismatch. | Verify cell morphology and the counterstain, compare a no-primary control, and repeat with an IHC-validated antibody if the pattern persists (general IHC practice). |
| A supposedly negative tissue still contains stained cells. | HPA's Not detected calls name particular cell types, such as adipocytes and cardiomyocytes, rather than every cell in their tissues (HPA: tissue IHC). | Identify the stained cell population before scoring a contradiction; investigate strong signal in the named negative cells with appropriate detection controls (HPA: tissue IHC; general IHC practice). |
| Weak staining in appendix, breast, or bronchus is scored as assay failure. | HPA reports Medium staining in the specified glandular or respiratory epithelial cells, below its High examples (HPA: tissue IHC). | Assess the stated cell population and cytoplasmic location, then compare a High-reference tissue processed in the same run before changing conditions (HPA: tissue IHC; general IHC practice). |
| Mitochondrial ICC-IF images appear inconsistent with diffuse cytoplasmic IHC. | The location summaries come from different assays: mitochondria in ICC-IF and cytoplasm in most tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). | For this chromogenic paraffin-section result, score the tissue-IHC cell and compartment pattern; keep the ICC-IF observation as assay-specific context (HPA: tissue IHC; HPA: subcellular ICC-IF). |
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.
| 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 → |
| 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 → |
Troubleshoot PRDX1 staining in paraffin sections by checking retrieval, compartment, controls, and cell-specific scoring before interpreting chromogenic signal.
Anti-PRDX1 antibodies have IHC images from human, mouse and rat tissues, plus IF images from human cells (catalog image captions).
PB9348 has IHC images from paraffin sections of human mammary cancer, mouse brain and rat brain, and IF images from HeLa and U20S cells (catalog image captions). M01845-3 has IHC images from rat kidney, human kidney and human thyroid; IF is listed as an application without a supplied IF image (catalog image captions; catalog applications).
Which to pick: For tissue IHC across human, mouse and rat, choose PB9348: its IHC captions document paraffin sections in all three species, with the fixative unreported (PB9348 IHC image captions). For IF/ICC, choose PB9348 because its application list includes both and its IF captions show human cell staining (PB9348 applications; PB9348 IF image captions). M01845-3 is a monoclonal alternative with listed human, mouse and rat reactivity and IHC/IF applications, but its IHC captions do not report tissue processing or fixative (M01845-3 catalog description; M01845-3 applications; M01845-3 reactivity; M01845-3 IHC image captions).