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PICK1 / PRKCA-binding protein · IHC design guide
Design Immunohistochemistry for PICK1
Plan chromogenic PICK1 IHC in paraffin sections using the catalog antibody’s documented workflow (datasheet A02457-2). Expect predominantly cytoplasmic staining, with high signal in endocrine glandular cells and CNS neurons (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PICK1 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 | Predominantly cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in endocrine glandular cells and CNS neurons (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02457-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Spleen+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | No specific expression regulator reported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified; no transmembrane segment (UniProt) |
Section 1
Recommended PICK1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with published chromogenic PICK1 staining methods for dewaxed sections (PMC8285178) and mouse frozen sections (PMC3038962).
| Sample | Paraffin-embedded mouse testis tissue; fixative not specified (datasheet A02457-2) |
| Fixation | Image fixative and duration unreported (datasheet A02457-2); 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 A02457-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02457-2) |
| Primary antibody | Rabbit anti-PICK1, 2-5 μg/ml (datasheet A02457-2) |
| Primary incubation | Overnight at 4 °C (datasheet A02457-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02457-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PICK1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression, most abundant in endocrine tissues and CNS. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A02457-2); citrate pH 6.0 was used on dewaxed sections in PMC8285178.
Section 2
What Is the Expected PICK1 Staining Pattern?
PICK1 staining should be predominantly cytoplasmic, with possible perinuclear or membrane-associated signal (UniProt Q9NRD5 localisation; UniProt Q9NRD5 topology: no transmembrane segment). Strong staining is documented in selected neuronal, endocrine glandular, muscle and testicular cells (HPA tissue IHC: High). HPA rates the tissue staining Approved, with medium consistency against RNA expression; use these patterns as guides rather than absolute tissue-wide rules (HPA tissue IHC reliability).
| Cytoplasmic staining is strong in hippocampal or cortical neurons and adrenal or parathyroid glandular cells (HPA tissue IHC: High). | This fits the reported cell and compartment pattern (HPA tissue IHC: neuronal and glandular cells High; UniProt Q9NRD5 localisation). Compare the relevant cells with background on the same section before scoring. |
| Perinuclear accentuation or fine cell-associated puncta accompany cytoplasmic staining (UniProt Q9NRD5 localisation). | These can fit PICK1's perinuclear, membrane-associated and synaptic locations (UniProt Q9NRD5 localisation). Chromogenic puncta alone do not identify a synapse; assess cellular context before assigning that structure. |
| Signal is exclusively nuclear, with no convincing cytoplasm (HPA tissue IHC: general cytoplasmic expression). | This conflicts with the reported pattern (HPA tissue IHC; UniProt Q9NRD5 localisation). Consider nonspecific staining or a reading artefact, and compare a known-positive section and detection controls before calling it PICK1. |
| Strong signal appears in spleen red-pulp cells or vaginal squamous epithelium (HPA tissue IHC: Not detected). | These cell populations were not detected in the HPA survey, so strong staining is discordant rather than definitive proof of cross-reactivity (HPA tissue IHC). Check tissue identity, nonspecific binding and chromogenic endogenous activity using appropriate controls (general IHC practice). |
| The whole section looks hazy, or a known-positive cell population has no signal (HPA tissue IHC: High examples). | Uniform haze limits compartment scoring; an absent signal in documented high-staining cells makes this run inconclusive (general IHC practice; HPA tissue IHC). Review negative and positive controls before inferring biological absence. |
💡Expected PICK1 appearanceCall a section positive when the relevant cells show predominantly cytoplasmic staining, strongest in documented High populations (HPA tissue IHC; UniProt Q9NRD5 localisation); isolated nuclear signal or uniform haze is suspect (general IHC practice).
| Tissue and cell selection | HPA reports High staining in neurons of caudate, cortex and hippocampus; adrenal and parathyroid glandular cells; skeletal myocytes; and spermatogonia (HPA tissue IHC). Appendix glandular cells are Medium, while several other listed populations are Low (HPA tissue IHC). Select controls by cell population, not tissue name alone. |
| Subcellular interpretation | PICK1 is cytoplasmic, perinuclear and membrane-associated, including at excitatory synapses; it has no transmembrane segment (UniProt Q9NRD5 localisation and topology). A cell-edge accent does not require a continuous membrane outline, and chromogenic IHC cannot establish synaptic identity from puncta alone (general IHC practice). |
| Evidence strength | The tissue IHC profile is Approved with medium staining–RNA consistency (HPA tissue IHC reliability). The listed IHC antibody HPA072674 is Approved; no Enhanced IHC status is supplied (HPA antibodies). Treat unexpected patterns as findings to verify, especially where staining is Low or Not detected (HPA tissue IHC). |
| Isoforms and epitope coverage | Two isoforms are listed, but the payload gives no antibody epitope or isoform-specific staining data (UniProt Q9NRD5 isoforms; HPA antibodies). The pattern cannot distinguish isoforms; use epitope documentation if an isoform-specific conclusion is needed. |
| IF/ICC Q&A: what localisation should IF show? | Supported HPA ICC-IF localisation is cytosol, with images listed for HAP1, HEK293 and U2OS (HPA subcellular ICC-IF). That observation supports a cytosolic IF expectation, but it does not validate an IHC-P condition or a synaptic-puncta claim (HPA subcellular ICC-IF; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells lack chromogenic signal (HPA tissue IHC: High examples). | A weak run, unsuitable detection conditions or tissue handling can make a positive control fail (general IHC practice). PICK1-specific fixation sensitivity is unreported in the supplied evidence. | Repeat with a documented High cell population and check section integrity, retrieval and detection controls using the antibody's IHC-P instructions (HPA tissue IHC; general IHC practice). |
| Staining appears only in nuclei (HPA tissue IHC: general cytoplasmic expression). | The compartment disagrees with reported cytoplasmic and perinuclear localisation (HPA tissue IHC; UniProt Q9NRD5 localisation); nonspecific signal or interpretation error is possible. | Recheck cell boundaries against the counterstain and compare positive and negative controls (general IHC practice). Do not score nuclear-only staining as the expected PICK1 pattern. |
| Strong signal appears in an HPA Not detected cell population (HPA tissue IHC). | Cross-reactivity, nonspecific binding or endogenous chromogenic activity is possible (general IHC practice); the HPA observation is not a universal biological negative. | Confirm the cell type; compare omission and detection controls, then repeat with an independently validated reagent if the discrepancy persists (general IHC practice). |
| Diffuse haze obscures cell boundaries (general IHC practice). | Excess background from staining or detection conditions can prevent compartment assignment (general IHC practice). | Inspect the negative control, blocking and washes; optimize staining conditions within the antibody's IHC-P guidance before comparing cell populations (general IHC practice). |
| A Low-staining tissue seems negative (HPA tissue IHC: Low populations). | Low reported staining may be hard to separate from background in that section (HPA tissue IHC; general IHC practice). | Check a High-staining control in the same run before reporting absence; score the specified cell population rather than the entire tissue (HPA tissue IHC; general IHC practice). |
| Puncta are interpreted as confirmed synapses (UniProt Q9NRD5 localisation). | PICK1 can occur at synapses, but chromogenic morphology alone lacks the resolution and markers to assign puncta to that compartment (UniProt Q9NRD5 localisation; general IHC practice). | Report the observed cytoplasmic or punctate pattern; reserve a synaptic assignment for an appropriately controlled colocalisation study (general IF practice). |
Experiment evidence
IHC & IF images showing PICK1 staining
A02457-2 · IHCIHC analysis of PICK1 using anti-PICK1 antibody (A02457-2). PICK1 was detected in a paraffin-embedded section of mouse testis tissue. Heat m
A02457-2 · IHCIHC analysis of PICK1 using anti-PICK1 antibody (A02457-2). PICK1 was detected in a paraffin-embedded section of rat brain tissue. Heat medi
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100151-PICK1/tissue/cerebral+cortex#imid_20062661
HPA · Parathyroid glandHuman Protein Atlas IHC, Parathyroid gland. Source: https://www.proteinatlas.org/ENSG00000100151-PICK1/tissue/parathyroid+gland#imid_20062544
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100151-PICK1/tissue/colon#imid_20062571
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100151-PICK1/tissue/liver#imid_20062588
HPA · ICC-IF HAP1Human Protein Atlas immunofluorescence, HAP1, antibody HPA000819; green = PICK1, blue = nucleus, red = microtubules.
HPA · ICC-IF HAP1Human Protein Atlas immunofluorescence, HAP1, antibody HPA000819; green = PICK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PICK1 IHC & IF
🧪Run cerebral cortex first and score neuronal cells, which show high PICK1 staining (HPA: High in cerebral cortex neuronal cells). Run spleen red pulp as the negative tissue (HPA: Not detected in spleen red-pulp cells); on the cortex slide, use adjacent non-neuronal cells as a background comparator, but do not assume they are PICK1-negative because expression is widespread (UniProt Q9NRD5: ubiquitous).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Spleen (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PICK1 in HAP1, HEK293, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, a rabbit IgG isotype control matched to the primary antibody where possible, and identically processed PICK1-knockout tissue as a biological negative (selected-SKU caption: rabbit primary antibody; standard IHC controls). Block endogenous peroxidase for DAB detection, and assess neuronal pigment as a possible source of background in cortex (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not report a fixative (selected-SKU tissue-IHC caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the captioned mouse testis section, but its necessity has not been established (selected-SKU tissue-IHC caption). For IF/ICC, HPA reports supported cytosolic localization and images from HAP1, HEK293 and U2OS; the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC (HPA: subcellular localization).
HPA tissue IHC evidence for PICK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced PICK1 IHC Tips
Troubleshooting chromogenic PICK1 IHC in paraffin sections, with one Q&A on IF/ICC.
Which retrieval condition should I try first for PICK1 in paraffin sections?
Start with heat-mediated retrieval in EDTA, pH 8.0, for paraffin-section PICK1 IHC (datasheet A02457-2). The selected mouse testis image used that condition, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A02457-2). If staining is weak, compare a modest range of heating durations while keeping section thickness, antibody concentration and detection settings constant (standard IHC practice). Include a no-primary section to identify retrieval-related background, and inspect tissue morphology alongside signal (standard IHC practice). Do not interpret a stronger diffuse stain alone as improved detection when PICK1 is expected mainly in the cytoplasm (HPA: general cytoplasmic expression).
How should I troubleshoot fixation when PICK1 staining is weak or uneven?
PICK1-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet A02457-2). Record the actual fixation and processing history for each sample, then compare sections processed together before changing the staining workflow (standard IHC practice). If a batch differs, assess tissue preservation and retrieval consistency on adjacent sections while holding the EDTA, pH 8.0 retrieval condition constant (datasheet A02457-2; standard IHC practice). Keep antibody concentration and detection time matched so changes in intensity can be interpreted (standard IHC practice). Avoid attributing weak PICK1 staining to a particular fixative without a controlled comparison.
Where should convincing PICK1 staining appear in chromogenic IHC?
Expect mainly cytoplasmic staining: tissue IHC describes general cytoplasmic expression, and PICK1 is also annotated at perinuclear regions, membranes and synapses (HPA: tissue profile; UniProt Q9NRD5: subcellular location). Interpret membrane-associated or punctate staining in its cellular context rather than requiring a continuous membrane rim; PICK1 has no transmembrane segment (UniProt Q9NRD5: topology). In testis, examine spermatogonia because HPA reports high staining there, while the selected antibody has a mouse testis paraffin-section image (HPA: testis; datasheet A02457-2). Compare nuclear-only or extracellular deposits with adjacent cell morphology and the no-primary control (standard IHC practice). A regional stain without identifiable positive cells warrants caution.
Could PICK1 isoforms or epitope access explain discordant staining?
PICK1 has 2 annotated isoforms and a PDZ domain at residues 22–105 plus an AH domain at 144–357 (UniProt Q9NRD5: isoforms and domains). The supplied antibody caption does not identify its epitope, so a difference between isoforms cannot be assigned to antibody recognition from these data (datasheet A02457-2). Check the antibody's documented immunogen or epitope against the isoforms before making an isoform claim (standard IHC validation practice). Keep retrieval and detection conditions matched when comparing tissues, because processing can change epitope accessibility (standard IHC practice). Treat a distinct staining pattern as a hypothesis until an independent specificity control supports it.
How can I adapt the PICK1 question to multiplex IF/ICC?
For IF/ICC, pair PICK1 with a marker of the cell population under study and compare signal within identified cells; HPA reports high PICK1 staining in neuronal cells of the cerebral cortex and hippocampus (HPA: cerebral cortex and hippocampus). Select a fluorophore channel after measuring autofluorescence in the specimen, and include single-stain controls to assess spectral bleed-through (standard IF practice). PICK1 is supported as cytosolic in ICC/IF and has no transmembrane segment, so use controlled permeabilisation when the epitope is intracellular (HPA: subcellular location; UniProt Q9NRD5: topology; standard IF practice). The supplied evidence does not specify the catalog antibody's IF epitope or fixation condition (datasheet A02457-2). Optimise those variables in IF/ICC rather than transferring settings from the paraffin-section image.
What should I change when PICK1 DAB staining is diffuse or widespread?
Run a no-primary control and block endogenous peroxidase before HRP detection to assess enzyme-related background (standard chromogenic IHC practice). The selected paraffin-section image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and DAB development (datasheet A02457-2). If background persists, compare lower primary concentrations or shorter incubation while keeping retrieval and detection conditions matched (standard IHC practice). Check whether deposits follow tissue edges, damaged areas or extracellular spaces rather than identifiable cytoplasm (standard IHC practice; HPA: general cytoplasmic expression). Because PICK1 expression is widespread, diffuse cellular staining requires specificity controls before it is dismissed as background (UniProt Q9NRD5: tissue specificity).
How should I score PICK1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because PICK1 tissue IHC is generally cytoplasmic and its abundance varies by cell type (HPA: tissue profile). For chromogenic sections, report the percentage of positive cells and an H-score combining staining intensity with that percentage, using identical thresholds across samples (standard IHC scoring practice). Alternatively, count positive cells per mm² of viable tissue when cell density is the biological readout (standard IHC scoring practice). Normalise to the number of eligible cells or viable tissue area, and record excluded folds, necrosis and section edges (standard IHC practice). Keep staining batches, imaging settings and region selection consistent across groups (standard IHC practice).
How can I distinguish genuine PICK1 staining from artefact?
A credible result places signal in identifiable cytoplasm, with any perinuclear or membrane-associated pattern judged against cell morphology (HPA: tissue profile; UniProt Q9NRD5: subcellular location). HPA reports high staining in testis spermatogonia and no detected staining in spleen red-pulp cells, which can guide comparison without establishing specificity for this antibody (HPA: testis and spleen). Be cautious with nuclear-only staining, extracellular DAB deposits, edge effects and necrotic areas (standard IHC interpretation; HPA: general cytoplasmic expression). Use a no-primary control to reveal endogenous enzyme or detection background, and compare adjacent intact tissue (standard IHC practice). An independent antibody or orthogonal assay strengthens a target-specific interpretation (standard IHC validation practice).
Boster reagents
Best PICK1 / PRKCA-binding protein IHC Antibodies
A02457-2 has real paraffin-section IHC images from mouse testis and rat brain (catalog IHC image captions). No IF/ICC image is supplied (catalog images).
Real IHC data
Anti-PICK1 Antibody ®
Cat # A02457-2
A02457-2 is the sole card and lists Human, Mouse, and Rat reactivity (catalog reactivity). Its IHC captions document mouse testis and rat brain paraffin sections; the rendered figure shows mouse testis (catalog IHC image captions; card figure).
Which to pick: Choose A02457-2 for mouse or rat paraffin-section tissue IHC: its captions use 2 μg/ml, within the listed 2–5 μg/ml IHC range (catalog IHC image captions; catalog dilution). The fixative is unreported (catalog IHC image captions). For IF/ICC or human tissue IHC, seek separate validation; IF/ICC is absent from the application list, and the IHC captions show only mouse and rat tissue despite listed Human reactivity (catalog applications; catalog IHC image captions; catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NRD5 (PICK1_HUMAN, PRKCA-binding protein).
- Human Protein Atlas. PICK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PICK1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. PICK1 antibody validation summary (2 antibodies).
- The TGF-β signalling negative regulator PICK1 represses prostate cancer metastasis to bone. British journal of cancer 2017 — PMC5572169.
- PICK1 Deficiency Exacerbates Sepsis-Associated Acute Kidney Injury. BioMed research international 2021 — PMC8285178.
- PICK1 deficiency impairs secretory vesicle biogenesis and leads to growth retardation and decreased glucose tolerance. PLoS biology 2013 — PMC3635866.
- Preserved acute pain and impaired neuropathic pain in mice lacking protein interacting with C Kinase 1. Molecular pain 2011 — PMC3038962.
- PubMed PMID:10623590 — UniProt-cited evidence.
- PubMed PMID:12529303 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PICK1 (PRKCA-binding protein). Expected localisation Predominantly cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02457-2). Validated IHC antibody: A02457-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Spleen. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse testis tissue; fixative not specified (datasheet A02457-2); 2. Fixation: Image fixative and duration unreported (datasheet A02457-2); 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 A02457-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02457-2); 8. Primary antibody: Rabbit anti-PICK1, 2-5 μg/ml (datasheet A02457-2); 9. Primary incubation: Overnight at 4 °C (datasheet A02457-2). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified; no transmembrane segment (UniProt); Regulation: No specific expression regulator reported (UniProt). Evidence: UniProt Q9NRD5, HPA tissue IHC · Approved, HPA ICC-IF · cytosol., Datasheet A02457-2, PMC8285178, PMC3038962. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PICK1/immunohistochemistry-ihc-guide.