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PINK1 / Serine/threonine-protein kinase PINK1, mitochondrial · IHC design guide
Design Immunohistochemistry for PINK1
Use adrenal glandular cells as a high-staining reference and bone marrow hematopoietic cells as an unstained reference (HPA tissue IHC). This paraffin IHC guide covers cytoplasmic scoring (HPA tissue IHC) and PINK1 accumulation after mitochondrial damage (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PINK1 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 tissue staining (HPA tissue IHC); mitochondrial pool (UniProt) | |
| Staining pattern | Cytoplasmic staining in adrenal glandular 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 |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M00201) | |
| Caveat | Tissue IHC reliability is uncertain (HPA tissue IHC) | |
| Regulation | Damage increases outer-membrane PINK1 (UniProt) | |
| Isoform / epitope | 2 isoforms; processing may alter epitope coverage (UniProt) |
Section 1
Recommended PINK1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published PINK1 staining methods for mouse lung, paraffin sections, and rat skin flaps (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. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen retrieval rule); compare published conditions when optimizing tissue-specific staining.
Section 2
What Is the Expected PINK1 Staining Pattern?
PINK1: mitochondria/cytosol, no TM segment (UniProt Q9BXM7); placental trophoblasts stain high (HPA: Uncertain).
| Cytoplasmic stain in placental trophoblasts (HPA: High). | Expected cell pattern; tissue IHC reliability is uncertain (HPA). |
| Nuclear-only staining (UniProt Q9BXM7). | Unexpected for PINK1; suspect artefact (UniProt Q9BXM7 localization). |
| Strong esophageal squamous staining (HPA: Not detected). | Unexpected; consider cross-reactivity or endogenous peroxidase activity (HPA: Not detected). |
| Diffuse stain across cells and stroma. | May reflect nonspecific background (general IHC practice). |
| No placental trophoblast signal (HPA: High). | Check assay controls; HPA rates the tissue IHC pattern uncertain (HPA). |
💡Expected PINK1 appearancePositive: high cytoplasmic trophoblast stain (HPA: High); nuclear-only signal is suspect (UniProt Q9BXM7).
| Mitochondrial damage (UniProt Q9BXM7) | Stalled import increases outer-membrane PINK1 (UniProt Q9BXM7). |
| Proteolytic processing (UniProt Q9BXM7) | Smaller PINK1 fragments localize mainly in cytosol (UniProt Q9BXM7). |
| IHC validation (HPA) | HPA001931 and CAB026191 are both rated Uncertain for IHC (HPA). |
| Situation | Likely cause | Next action |
|---|---|---|
| Placental trophoblasts are blank (HPA: High). | Assay failure is possible; HPA tissue IHC reliability is uncertain (HPA). | Check a positive control and review retrieval (general IHC practice). |
| Signal is nuclear only (UniProt Q9BXM7). | Nuclear localization is unexpected (UniProt Q9BXM7). | Check the no-primary control and antibody specificity (general IHC practice). |
| Esophageal squamous cells stain (HPA: Not detected). | Cross-reactivity or endogenous peroxidase activity is possible (general IHC practice). | Check the no-primary control and peroxidase block (general IHC practice). |
| Diffuse section-wide haze appears. | Nonspecific binding or detection background is possible (general IHC practice). | Optimize blocking, antibody dilution and washes (general IHC practice). |
| Cytoplasmic patterns vary between samples. | Processing and mitochondrial damage affect PINK1 location (UniProt Q9BXM7). | Record the observed compartment and score intensity consistently. |
| IF/ICC pattern is unclear. | No ICC-IF images or main location are available (HPA: subcellular). | Assess colocalization with a mitochondrial marker (UniProt Q9BXM7 localization). |
Experiment evidence
IHC & IF images showing PINK1 staining
M00201 · IHCImmunohistochemical analysis of paraffin-embedded H. stomach section using Pink1(115-213). M00201 was diluted at 1:25 dilution. A undiluted
M00201 · IHCImmunohistochemical analysis of paraffin-embedded H. heart section using Pink1(115-213). M00201 was diluted at 1:25 dilution. A undiluted bi
M00201 · IFFluorescent image of PC12 cells stained with Pink1 (115-213) Antibody. M00201 was diluted at 1:25 dilution. An Alexa Fluor® 488-conjugated g
A00201 · IHCImmunohistochemistry of PINK1 in mouse brain tissue with PINK1 Antibody at 5 μg/mL.
A00201 · IFImmunofluorescence of PINK1 in mouse brain tissue with PINK1 Antibody at 20 μg/mL. Green: PINK1 antibody (A00201) Red: Phylloidin staining B
A00201-2 · IFIF analysis of PINK1 using anti-PINK1 antibody (A00201-2). PINK1 was detected in immunocytochemical section of A431 cells. Enzyme antigen re
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000158828-PINK1/tissue/cerebral+cortex#imid_1047980
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000158828-PINK1/tissue/colon#imid_1047914
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000158828-PINK1/tissue/liver#imid_1047915
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000158828-PINK1/tissue/kidney#imid_1047900
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PINK1 IHC & IF
🧪Run placenta first and expect trophoblastic cells to stain (High; HPA: placenta, trophoblastic cells). Use oral mucosa squamous epithelium as the negative tissue (Not detected; HPA: oral mucosa, squamous epithelial cells); on the placenta slide, adjacent unstained cells can serve as a background reference, but their PINK1-negative status is unverified (HPA: placenta, trophoblastic cells High).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for PINK1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and a concentration-matched control appropriate to the primary: host- and subclass-matched isotype for a monoclonal antibody, or host-matched nonimmune IgG for a polyclonal antibody (standard IHC practice). Use PINK1 knockout material as a biological specificity control; for chromogenic placenta sections, quench endogenous peroxidase and check for blood-associated background (standard IHC practice).
⚠️Feasibility: The selected M00201 caption documents paraffin-section IHC on stomach at 1:25, but its fixative is unreported; no target-specific fixation window or fixation effect is supplied (M00201 tissue-IHC caption). Antigen-retrieval dependence is unreported, so optimize retrieval empirically with the IHC-validated antibody (M00201 tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC; in placenta, blood-associated peroxidase can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for PINK1
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.
Positive expression · recommended positive controls
| 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 → |
Undetected expression · recommended negative controls
| 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 → |
Section 3
Advanced PINK1 IHC Tips
Troubleshoot PINK1 staining in paraffin sections by checking retrieval, compartment pattern, cell identity and controls before interpreting chromogenic signal.
How should I retrieve PINK1 antigen when paraffin sections stain weakly?
Use citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule: cytoplasmic or membrane antigen). The selected M00201 image documents staining in a paraffin stomach section at 1:25, but gives no retrieval conditions, so treat the page setting as a starting condition rather than a product-tested result (M00201 tissue-IHC caption). If signal remains weak, compare a second retrieval condition on adjacent sections while holding antibody concentration, detection and development time constant (standard IHC practice). Check tissue integrity and background alongside signal, because excessive retrieval can make a brighter section less interpretable (standard IHC practice).
Could fixation explain weak or uneven PINK1 staining?
Target-specific fixation sensitivity is unknown: the selected M00201 caption identifies a paraffin section but does not state its fixative (M00201 tissue-IHC caption). Record the actual fixative and fixation time for each specimen, then compare similarly processed sections before attributing a difference to PINK1 biology (standard IHC practice). When troubleshooting, stain adjacent sections from the same block together and inspect morphology, section attachment and staining gradients; these comparisons help reveal processing variation (standard IHC practice). Do not infer fixation tolerance from tissue staining patterns or from PINK1 localisation and phosphorylation, since those sources do not test fixation effects (HPA tissue IHC; UniProt Q9BXM7; M00201 tissue-IHC caption).
Where should a credible PINK1 signal appear in tissue sections?
Assess cytoplasmic staining in the relevant cells first: HPA describes a general cytoplasmic tissue-IHC profile, although its reliability is uncertain (HPA tissue IHC). PINK1 can occupy mitochondrial outer and inner membranes, while smaller processed fragments localise mainly to cytosol; damaged, depolarised mitochondria can accumulate PINK1 on their outer membrane (UniProt Q9BXM7 localisation). In chromogenic sections, a diffuse cytoplasmic pattern therefore does not by itself prove mitochondrial localisation or damage-induced accumulation (UniProt Q9BXM7 localisation; standard IHC interpretation). Compare cellular distribution with morphology and controls, and investigate dominant nuclear, extracellular or section-edge signal as possible nonspecific staining (standard IHC practice).
Could epitope choice change what my PINK1 stain represents?
Confirm the immunogen and epitope documented for the antibody in use; the selected caption names Pink1(115-213), but does not map individual isoform recognition (M00201 tissue-IHC caption). PINK1 has 2 annotated isoforms, a kinase domain at residues 156–511, and an annotated chain beginning at residue 78 (UniProt Q9BXM7). Processing produces smaller fragments that localise mainly to cytosol, so staining from an antibody against a retained epitope may represent more than one molecular form (UniProt Q9BXM7 processing and localisation). Do not infer which form dominates a section from staining intensity alone; compare antibody documentation and a suitable specificity control before assigning an isoform (standard IHC interpretation).
How can IF help assess the cell type and compartment behind IHC staining?
On a matched section, multiplex PINK1 with a marker identifying the expected cell type, chosen from the tissue and question being studied; HPA reports high staining in placental trophoblastic cells, for example (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and place the weaker channel away from strong tissue autofluorescence, using single-stain and unstained controls to assess bleed-through and background (standard IF practice). For an epitope exposed to cytosol, optimise permeabilisation to permit antibody access while preserving mitochondrial structure; confirm the antibody epitope before choosing conditions (UniProt Q9BXM7 localisation; standard IF practice). The supplied HPA subcellular record provides no ICC/IF images, so assess IF performance independently of the paraffin IHC caption (HPA subcellular; M00201 tissue-IHC caption).
How do I reduce diffuse brown staining without losing PINK1 signal?
Run a section without primary antibody to measure detection-system background, and block endogenous peroxidase before DAB development as part of the chromogenic workflow (standard IHC practice). The selected M00201 image used a 1:25 antibody dilution, an undiluted biotinylated secondary and DAB, so that dilution describes one image condition rather than an established optimum for every section (M00201 tissue-IHC caption). If staining is diffuse, titrate primary antibody and secondary detection separately while keeping retrieval and DAB development consistent (standard IHC practice). Inspect pigment and tissue edges under the microscope, and assess whether brown deposits remain in the no-primary control before calling them PINK1-positive cells (standard IHC practice).
What should I score when comparing PINK1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the target cell population and score cytoplasmic signal using the percentage of positive cells and an H-score, applying the same intensity thresholds across sections (HPA tissue IHC; standard IHC scoring practice). Record the denominator as evaluable cells of that type, or report positive-cell density per mm² of viable tissue when cell counting is impractical (standard IHC scoring practice). Normalise comparisons to the same cell population, viable area and staining batch, with a reference section included across batches (standard IHC scoring practice). Report cell-specific results because HPA's tissue profile has uncertain reliability and includes both high-staining populations and populations where staining was not detected (HPA tissue IHC).
How can I distinguish genuine PINK1 staining from an artefact?
A credible result should occur within identifiable cells and fit a cytoplasmic pattern; HPA reports high staining in placental trophoblastic cells but rates overall tissue-IHC reliability as uncertain (HPA tissue IHC). Compare the suspect area with a no-primary control and adjacent intact tissue, excluding staining confined to section edges, necrotic material or endogenous enzyme activity (standard IHC practice). Treat a predominantly nuclear or extracellular pattern as requiring further validation because the annotated PINK1 locations are mitochondrial membranes and cytosol (UniProt Q9BXM7 localisation). Even convincing cytoplasmic DAB staining does not establish kinase activation or mitophagy; those interpretations require evidence beyond PINK1 abundance and location in a single section (UniProt Q9BXM7 function; standard IHC interpretation).
Boster reagents
Best PINK1 / Serine/threonine-protein kinase PINK1, mitochondrial IHC Antibodies
Anti-PINK1 antibodies have IHC images from human stomach and mouse brain, plus an IF/ICC image from A431 cells (M00201, A00201 and A00201-2 image captions).
Real IHC data
Anti-PINK1 Antibody
Cat # M00201
Real IHC data
Anti-PINK1 Antibody
Cat # A00201
Real IF data
Anti-PINK1 Antibody ®
Cat # A00201-2
M00201 shows IHC in paraffin-embedded human stomach, and A00201 shows IHC in mouse brain (respective IHC image captions). A00201-2 shows IF/ICC in A431 cells (A00201-2 IF image caption).
Which to pick: For paraffin-section IHC, choose mouse monoclonal M00201 for human tissue at 1:25; its stomach caption reports paraffin embedding but leaves the fixative unreported (M00201 catalog entry and IHC image caption). For human-cell IF/ICC, choose A00201-2 at 5 μg/mL (A00201-2 application list and IF image caption). For a cross-species project, consider rabbit A00201, listed for human, mouse and rat at an IHC starting concentration of 5 μg/mL; its IHC image documents mouse brain only, with processing and fixative unreported (A00201 catalog entry and IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BXM7 (PINK1_HUMAN, Serine/threonine-protein kinase PINK1, mitochondrial).
- Human Protein Atlas. PINK1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. PINK1 subcellular location (ICC-IF): Highest expression in SiHa: 82.2 nTPM.
- Human Protein Atlas. PINK1 antibody validation summary (2 antibodies).
- Complement protein C1q induces endothelial apoptosis through C1qBP regulation of mitochondrial function in lung endothelial cells in systemic sclerosis. Molecular medicine (Cambridge, Mass.) 2026 — PMC13069790.
- PINK1 Immunoexpression Predicts Survival in Patients Undergoing Hepatic Resection for Colorectal Liver Metastases. International journal of molecular sciences 2023 — PMC10095114.
- TRPC6-Calpain-1 Axis Promotes Tubulointerstitial Inflammation by Inhibiting Mitophagy in Diabetic Kidney Disease. Kidney international reports 2024 — PMC11551102.
- Activation of aldehyde dehydrogenase-2 improves ischemic random skin flap survival in rats. Frontiers in immunology 2023 — PMC10335790.
- PubMed PMID:11494141 — UniProt-cited evidence.
- PubMed PMID:14607334 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PINK1 (Serine/threonine-protein kinase PINK1, mitochondrial). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial pool (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: M00201 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Bone marrow. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded H. stomach tissue; fixative not specified (datasheet M00201); 2. Fixation: Image fixative and duration unreported (datasheet M00201); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Mouse monoclonal (clone 38CT20.8.5) anti-PINK1, 1:25 (datasheet M00201); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M00201); Caveat: Tissue IHC reliability is uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; processing may alter epitope coverage (UniProt); Regulation: Damage increases outer-membrane PINK1 (UniProt). Evidence: UniProt Q9BXM7, HPA tissue IHC · Uncertain, HPA cell-line RNA · Highest expression in SiHa: 82.2 nTPM, Datasheet M00201, PMC13069790, PMC11551102. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PINK1/immunohistochemistry-ihc-guide.