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- Table of Contents
Plan PARK7 chromogenic IHC on paraffin sections with the catalog antibody at 2–5 µg/ml (datasheet M00757-4). Use cytoplasmic and nuclear tissue staining as the scoring reference (HPA tissue IHC), while accounting for stress-related changes in localisation (UniProt).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Cytoplasmic and nuclear in most tissues (HPA tissue IHC) | |
| Staining pattern | Glandular and other cells show cytoplasmic and nuclear staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00757-4) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 44 tissues); use no-primary + isotype controls |
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Oxidative stress can shift staining toward nuclei (UniProt) | |
| Regulation | No expression-level regulator specified (UniProt) | |
| Isoform / epitope | 0 annotated isoforms; intracellular epitope expected (UniProt) |
The catalog antibody’s IHC-P protocol (datasheet: M00757-4) is followed by three published PARK7 IHC protocols (PMC6377712; PMC3260266; PMC11538469).
| Sample | Paraffin-embedded human endometrial carcinoma tissue; fixative not specified (datasheet M00757-4) |
| Fixation | Image fixative and duration unreported (datasheet M00757-4); 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 M00757-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00757-4) |
| Primary antibody | Mouse monoclonal (clone 4B10) anti-PARK7, 2-5 µg/ml (datasheet M00757-4) |
| Primary incubation | Overnight at 4 °C (datasheet M00757-4) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00757-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PARK7-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in most tissues. No signal in the no-primary control. |
PARK7 staining should be predominantly cytoplasmic, with nuclear signal in many tissues (HPA: cytoplasmic and nuclear expression in most tissues; UniProt Q99497: predominantly cytoplasmic under normal conditions). High staining is reported in several glandular populations, hippocampal glia, and lung alveolar type I cells (HPA tissue IHC). Interpret intensity by cell type: HPA rates tissue IHC reliability Supported, with medium consistency between staining and RNA data (HPA tissue IHC). PARK7 has no transmembrane segment (UniProt Q99497 topology).
| Cytoplasmic staining with some nuclear staining in an expected cell population. | This fits the broad tissue pattern (HPA tissue IHC) and the normally predominant cytoplasmic localisation (UniProt Q99497). Judge the relevant cells, rather than the entire section, when calling a positive result (general IHC practice). |
| A sharply membrane-only or extracellular pattern, without credible cytoplasmic staining. | Treat this as a possible artefact and check controls before assigning PARK7: it lacks a transmembrane segment, and its usual location is cytoplasmic (UniProt Q99497). Reported membrane raft localisation does not establish a membrane-only IHC pattern (UniProt Q99497). |
| Strong staining confined to an unexpected cell type while expected cells stay faint. | Investigate cross-reactivity or endogenous detection activity (general IHC practice). HPA reports high staining in defined cell populations, such as hippocampal glia and pancreatic exocrine glandular cells; those observations do not establish that every cell in those tissues should be high (HPA tissue IHC). |
| Diffuse colour over tissue and empty areas, obscuring cell boundaries. | Interpret this as background until a clean cellular pattern is visible (general IHC practice). Compare a no-primary control and assess blocking, detection reagents, and washing; diffuse colour alone cannot establish PARK7 localisation (general IHC practice). |
| No discernible signal in a tissue containing an HPA high-staining cell population. | Check the sampled cell population and the IHC run before calling the tissue negative (general IHC practice). HPA reports high staining in, for example, adrenal glandular cells and duodenal Brunner glands, but rates overall tissue IHC reliability Supported with medium consistency (HPA tissue IHC). |
| Cell population and tissue | HPA rates adrenal, breast, epididymal, and fallopian tube glandular cells High, alongside duodenal Brunner glands, hippocampal glia, lung alveolar type I cells, and pancreatic exocrine glandular cells (HPA tissue IHC). Its Low calls include cardiomyocytes and smooth muscle cells (HPA tissue IHC). |
| Compartment and cellular state | Cytoplasm predominates normally; UniProt reports movement to mitochondria and then nucleus with oxidative stress (UniProt Q99497). A changed compartment could therefore have biological context, but staining alone does not establish oxidative stress; assess morphology and controls (general IHC practice). |
| Evidence across sources | UniProt reports high expression in heart and pancreatic islets above surrounding exocrine tissue (UniProt Q99497); HPA rates cardiomyocyte staining Low and pancreatic exocrine glandular staining High (HPA tissue IHC). These are source-specific observations; do not convert them into a universal IHC intensity ranking. |
| Antibody validation | HPA lists IHC as Supported for HPA004190 and CAB005870; HPA004190 also has ICC Supported (HPA antibodies). Supported tissue IHC has medium consistency with RNA data (HPA tissue IHC). Use the validation status as context, not proof that every signal is specific (general IHC practice). |
| IF/ICC Q&A: what localisation should IF show? | Mainly nucleoplasm and cytosol; flagellar centriole and mid piece are additional reported locations (HPA ICC-IF). These IF observations help check compartment plausibility, but they do not supply an IHC-P protocol or make the additional locations expected in every cell (HPA ICC-IF). |
| Fixation sensitivity | Target-specific effects of fixation on PARK7 staining are unreported in the supplied UniProt and HPA evidence. Record the preparation and compare matched controls when investigating a result (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular cells show no stain. | The expected population may be absent from the section, or the IHC run may have failed (general IHC practice). | Confirm cell identity on the counterstained section, then inspect a tissue control containing an HPA High population and the run controls (HPA tissue IHC; general IHC practice). |
| Nuclei stain but cytoplasm is consistently blank. | That pattern is incomplete for normally predominant cytoplasmic PARK7; nuclear localisation can occur, so the cause is unresolved from appearance alone (UniProt Q99497). | Compare an expected positive and detection controls; review antibody dilution and antigen retrieval against the catalog antibody's IHC-P instructions (general IHC practice). |
| Colour is strongest along membranes or tissue edges. | An edge or detection artefact is possible; PARK7 has no transmembrane segment (UniProt Q99497 topology; general IHC practice). | Compare interior cells with edges, inspect a no-primary control, and reassess blocking and washing before scoring (general IHC practice). |
| Many unrelated cell types stain strongly. | The pattern may reflect nonspecific binding or endogenous detection activity; HPA High calls apply to specified cell populations (general IHC practice; HPA tissue IHC). | Inspect no-primary and appropriate detection controls, then adjust blocking or detection conditions as indicated by those controls (general IHC practice). |
| A low-staining HPA cell population appears negative. | Low means weak staining is reported, so absence of visible colour at one exposure or threshold is inconclusive (HPA tissue IHC; general IHC practice). | Check an HPA High cell population in the same run before interpreting the low-staining cells, and retain the HPA reliability caveat (HPA tissue IHC; general IHC practice). |
| IHC and IF appear to disagree about nuclear signal. | HPA tissue IHC reports cytoplasmic and nuclear expression, while ICC-IF supports nucleoplasm and cytosol; samples and assay readouts differ (HPA tissue IHC; HPA ICC-IF). | Compare cell identity and controls within each assay, then report each observed compartment with its method instead of inferring that one image invalidates the other (general IHC/IF practice). |
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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glands of Brunner | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: PARK7 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Troubleshoot PARK7 staining by checking retrieval, compartment, cell type and controls before comparing chromogenic signal across sections (datasheet M00757-4; HPA tissue IHC; UniProt Q99497).
Anti-PARK7 antibodies have IHC images from human tumor and mouse pancreas paraffin sections, plus IF/ICC images from HeLa and SK-OV-3 cells (catalog image captions).
M00757 has a human bladder cancer paraffin-section IHC image; M00757-4 has human endometrial carcinoma paraffin-section IHC and HeLa IF/ICC images (respective catalog image captions). PB9308 has mouse pancreas IHC and SK-OV-3 IF/ICC images; its catalog also lists human and rat reactivity (PB9308 image captions; catalog reactivity).
Which to pick: For human tissue IHC, choose M00757-4 when a documented starting condition helps: its paraffin-section caption reports EDTA retrieval at pH 8.0 and 2 µg/ml primary antibody; the fixative is unreported (M00757-4 IHC caption). For IF/ICC, M00757-4 has a HeLa image and PB9308 has a SK-OV-3 image (respective IF captions). For cross-species tissue IHC, PB9308 lists human, mouse and rat reactivity and shows mouse and rat pancreas IHC; M00757 is a rabbit monoclonal with listed human, mouse and rat reactivity and a human bladder cancer IHC image (PB9308 catalog reactivity and IHC captions; M00757 catalog host, clone, reactivity and IHC caption).