This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
Plan paraffin-section PINK1 staining around a generally cytoplasmic tissue profile with uncertain reliability (HPA tissue IHC). Start with the IHC-validated antibody M00201 at 1:25 and compare staining with matched controls (datasheet: IHC-P 1:25; standard IHC practice).
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); mitochondrial PINK1 (UniProt) | |
| Staining pattern | Cytoplasmic staining in glandular and trophoblastic cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining reliability is uncertain (HPA tissue IHC) | |
| Regulation | Damage stabilizes outer-membrane PINK1 (UniProt) | |
| Isoform / epitope | 2 isoforms; processing warrants epitope mapping (UniProt) |
The catalog antibody’s IHC-P protocol is followed by three published PINK1 IHC protocols from paraffin sections (PMC13069790; PMC11551102; PMC10335790).
| Sample | Paraffin-embedded H. stomach tissue; fixative not specified (datasheet M00201) |
| Fixation | Image fixative and duration unreported (datasheet M00201); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Mouse monoclonal (clone 38CT20.8.5) anti-PINK1, 1:25 (datasheet M00201) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PINK1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
PINK1 localizes mainly to mitochondria, while smaller processed fragments localize mainly to cytosol; its UniProt record lists outer and inner mitochondrial membranes but no transmembrane segment (UniProt Q9BXM7 localization and topology). In paraffin tissue, expect predominantly cytoplasmic staining in HPA positive cell populations, especially placental trophoblasts and adrenal glandular cells (HPA tissue IHC). HPA rates the tissue pattern Uncertain, so interpret intensity cautiously (HPA tissue IHC).
| Cytoplasmic chromogen in placental trophoblasts or adrenal glandular cells, with little signal in adjacent unstained structures (HPA tissue IHC). | This fits HPA's general cytoplasmic profile and its High ratings for those cell types (HPA tissue IHC). Compare cells within the same section; the High rating does not guarantee uniform staining in every specimen (HPA tissue IHC; standard IHC practice). |
| Predominantly nuclear staining, without a convincing cytoplasmic component (HPA tissue IHC; UniProt Q9BXM7 localization). | Nuclear localization is absent from the supplied PINK1 record and HPA describes general cytoplasmic expression (UniProt Q9BXM7 localization; HPA tissue IHC). Treat a nuclear-only pattern as suspect and check the no-primary control and antibody staining conditions (standard IHC practice). |
| Strong staining in cells HPA lists as Not detected, such as bone marrow hematopoietic cells or esophageal squamous epithelium (HPA tissue IHC). | Consider cross-reactivity or endogenous detection activity, especially if expected positive cells are weak (standard IHC practice; HPA tissue IHC). A Not detected HPA result is an assay observation, not proof that every specimen must be blank (HPA tissue IHC). |
| Broad, diffuse color across cells and surrounding tissue, with little distinction between cellular compartments (standard IHC practice). | This is difficult to score as PINK1's reported cytoplasmic pattern (HPA tissue IHC). Check the no-primary control, blocking, washing and chromogen development before assigning biological meaning (standard IHC practice). |
| No chromogen in an HPA High cell population, including placental trophoblasts or rectal glandular cells (HPA tissue IHC). | First assess section integrity, retrieval, antibody dilution and detection controls (standard IHC practice). Because both listed HPA antibodies have Uncertain IHC validation, an absent result alone cannot establish absent PINK1 protein (HPA antibody validation). |
| Localization and processing (UniProt Q9BXM7 localization). | PINK1 is mainly mitochondrial, while two smaller proteolytic fragments localize mainly to cytosol (UniProt Q9BXM7 localization). A cytoplasmic IHC signal need not resolve individual mitochondria; compare its cellular distribution with the HPA tissue profile (HPA tissue IHC; standard IHC practice). |
| Tissue evidence differs by measurement (UniProt Q9BXM7 tissue specificity; HPA tissue IHC). | UniProt reports high expression in skeletal muscle, yet HPA records skeletal myocytes as Not detected by tissue IHC and RNA specificity as enhanced in skeletal muscle and tongue (UniProt Q9BXM7 tissue specificity; HPA tissue IHC). Do not use skeletal muscle as an assured IHC positive control (HPA tissue IHC). |
| Strength of IHC evidence (HPA tissue IHC; HPA antibody validation). | HPA rates the overall tissue pattern Uncertain because antibody staining and RNA data have medium consistency; both listed antibodies also have Uncertain IHC status (HPA tissue IHC; HPA antibody validation). Use morphology and appropriate controls before interpreting weak or unexpected staining (standard IHC practice). |
| IF/ICC Q&A: Is a specific fluorescent PINK1 pattern established here? (HPA subcellular ICC-IF). | HPA provides no main subcellular location or ICC-IF image cell lines in this payload, so it does not establish an observed IF pattern (HPA subcellular ICC-IF). UniProt supports mitochondrial and cytosolic localization as biological context, not an IF validation result (UniProt Q9BXM7 localization). |
| Situation | Likely cause | Next action |
|---|---|---|
| An HPA High population is blank (HPA tissue IHC). | Insufficient detection, unsuitable retrieval or antibody dilution, or a section problem may suppress visible chromogen (standard IHC practice). | Check tissue preservation and a functioning detection control; review the antibody's documented IHC-P conditions before adjusting retrieval or dilution (standard IHC practice). Interpret a persistently blank result in light of HPA's Uncertain rating (HPA tissue IHC). |
| The no-primary control develops color (standard IHC practice). | Endogenous enzyme activity or detection reagent background can produce chromogen independently of primary antibody (standard IHC practice). | Address endogenous activity for the chosen chromogenic system, reassess blocking and washes, then repeat the no-primary control alongside the specimen (standard IHC practice). |
| The entire section has hazy cytoplasmic color (standard IHC practice). | Nonspecific binding or excessive chromogen development can obscure cell-level differences (standard IHC practice). | Compare with the no-primary control and review blocking, antibody dilution, wash stringency and development time; score only a reproducible cellular pattern (standard IHC practice). |
| Staining is mainly nuclear (HPA tissue IHC; UniProt Q9BXM7 localization). | The observed compartment conflicts with HPA's general cytoplasmic profile and the supplied UniProt localization (HPA tissue IHC; UniProt Q9BXM7 localization). | Recheck morphology and counterstain, compare a known HPA High cell population, and inspect controls before assigning the nuclear signal to PINK1 (HPA tissue IHC; standard IHC practice). |
| A listed Not detected population stains strongly (HPA tissue IHC). | Cross-reactivity or endogenous chromogenic activity is possible; HPA's Uncertain reliability also limits the reference pattern (standard IHC practice; HPA tissue IHC). | Compare the no-primary control and a High reference population on the same run, then report the discrepancy without treating either result as definitive (standard IHC practice; HPA tissue IHC). |
| Skeletal myocytes show no signal despite expected expression from UniProt (UniProt Q9BXM7 tissue specificity). | HPA tissue IHC lists skeletal myocytes as Not detected, despite UniProt's high tissue expression description (HPA tissue IHC; UniProt Q9BXM7 tissue specificity). | Use an HPA High IHC population, such as placental trophoblasts, to assess the staining run; document the skeletal muscle discrepancy rather than changing the scoring threshold to force positivity (HPA tissue IHC; standard IHC practice). |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 | Myoepithelial cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Rectum | Glandular cells | High | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Troubleshoot PINK1 chromogenic IHC in paraffin sections using the catalog antibody’s tissue image, reported localisation, and appropriate staining controls.
Anti-PINK1 antibodies have IHC images from human paraffin stomach and heart sections and mouse brain, plus an IF/ICC image from A431 cells (catalog image captions).
M00201 has human paraffin stomach and heart IHC images; A00201 has mouse brain IHC and IF images (catalog image captions). A00201-2 lists human IF/ICC and has an A431 cell IF image (catalog applications/reactivity; A00201-2 IF caption).
Which to pick: For human tissue IHC-P, choose M00201, a mouse monoclonal: its own captions show paraffin stomach and heart sections stained with DAB; the fixative is unreported (catalog clone/applications; M00201 IHC captions). For human-cell IF/ICC, choose A00201-2, which lists both applications and has an A431 cell IF image (catalog applications/reactivity; A00201-2 IF caption). For cross-species planning, A00201 lists human, mouse and rat reactivity and IHC-P/IF applications, with mouse brain images for both; its IHC caption does not report processing or fixative (catalog applications/reactivity; A00201 image captions).