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PICALM / Phosphatidylinositol-binding clathrin assembly protein · IHC design guide
Design Immunohistochemistry for PICALM
Plan paraffin section staining for PICALM using the IHC-validated antibody and its documented dilution of 0.5–1 μg/mL (datasheet A02053-1). Compare cytoplasmic and membranous staining with tissue controls informed by the Human Protein Atlas profile (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PICALM 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic/membranous; highest in GI, kidney, placenta (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02053-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Skeletal muscle+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium concordance (HPA tissue IHC) | |
| Regulation | Broad expression across examined tissues (UniProt) | |
| Isoform / epitope | 5 isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended PICALM IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A02053-1) with four published PICALM IHC methods (PMC4148152; PMC3949918; PMC9063212; PMC4482781).
| Sample | Paraffin-embedded human Ovarian cancer tissue; fixative not specified (datasheet A02053-1) |
| Fixation | Image fixative and duration unreported (datasheet A02053-1); 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 A02053-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02053-1) |
| Primary antibody | Rabbit anti-PICALM, 0.5-1μg/ml (datasheet A02053-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02053-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A02053-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PICALM-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic and membranous expression, most abundant in gastrointestinal tract, kidney and placenta. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet: A02053-1); citrate retrieval is documented for other antibodies (PMC4148152; PMC3949918; PMC9063212).
Section 2
What Is the Expected PICALM Staining Pattern?
PICALM should appear mainly in cytoplasmic and membranous compartments, consistent with its coated-pit and vesicle localisation and lack of a transmembrane segment (UniProt Q13492). Expect broad tissue staining, with prominent gastrointestinal, kidney and placental expression (HPA: tissue IHC). Interpret intensity by cell type: several glandular and endocrine populations stain strongly, while muscle cells may be unstained (HPA: tissue IHC). HPA rates the tissue pattern Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC).
| Cytoplasmic and membranous signal in glandular or endocrine cells, with nearby tissue structures distinguishable. | This fits the reported broad pattern; strong glandular staining occurs in adrenal gland, breast, endometrium, fallopian tube and gallbladder, while endocrine cells stain strongly in appendix, colon and duodenum (HPA: tissue IHC). Judge the relevant cell population rather than treating every cell in a section as an equal positive control. |
| Predominantly nuclear staining, or a uniform outline of every cell with little cytoplasmic detail. | A nuclear-only or uniformly outlined pattern warrants an artefact check against the expected cytoplasmic and membranous tissue pattern (HPA: tissue IHC). Nuclear localisation is possible in some cells through interaction with PIMREG, so isolated nuclear signal is not sufficient by itself to establish nonspecific staining (UniProt Q13492). |
| Strong chromogen in skeletal myocytes or smooth muscle cells, especially when expected positive cells are weak. | These cell types are reported as not detected in the sampled HPA tissues (HPA: tissue IHC). Check cell identification and compare a no-primary control: unexpected colour may reflect cross-reactivity or, with enzyme-based detection, endogenous activity (standard IHC practice). A single discrepant field cannot establish either cause. |
| Diffuse colour across cells and extracellular areas, obscuring compartment boundaries. | That distribution does not resolve the reported cytoplasmic and membranous pattern (HPA: tissue IHC). Review the no-primary control, wash and blocking steps, and chromogen development (standard IHC practice). Interpret apparent low-level positivity only after background is sufficiently controlled to identify individual cells. |
| No detectable signal in an HPA-reported high-staining glandular or endocrine population. | First verify that the expected cells are present in the section; high staining refers to specified cell types, not every structure in the named organ (HPA: tissue IHC). If those cells are present, review the IHC-P detection workflow and antibody use before calling the sample biologically negative (standard IHC practice). |
💡Expected PICALM appearanceA convincing positive shows identifiable cytoplasmic and membranous staining in the relevant glandular or endocrine cells, potentially strong in the HPA-listed high-staining populations (HPA: tissue IHC); diffuse colour without cell boundaries or dominant staining of reported negative myocytes is suspect (HPA: tissue IHC; standard IHC practice).
| Tissue and cell selection | PICALM is expressed across examined tissues (UniProt Q13492), but HPA tissue IHC varies by cell population: high staining is documented in selected glandular and endocrine cells, whereas skeletal and smooth muscle cells are reported as not detected (HPA: tissue IHC). Choose controls by the named cell population and assess that population on the slide. |
| Compartment and topology | PICALM is associated with the cell membrane, coated pits, Golgi area and clathrin-coated vesicles, and has no transmembrane segment (UniProt Q13492). Its membrane-associated signal need not resemble a continuous surface rim; read punctate or regional cytoplasmic staining alongside the tissue pattern (UniProt Q13492; HPA: tissue IHC). |
| Interpretation across antibodies | HPA lists both HPA019053 and HPA019061 as IHC Enhanced (HPA: antibody validation). The tissue profile is also rated Enhanced, but its staining and RNA data have medium consistency (HPA: tissue IHC). Treat a matching pattern as supporting evidence, while checking unexpected cell types or compartments against controls. |
| Isoforms and epitope coverage | UniProt records five PICALM isoforms and an ENTH domain at residues 14–145 (UniProt Q13492). The supplied evidence does not map the catalog antibody epitope or establish isoform-specific IHC staining. Do not attribute a tissue difference to an isoform, phosphorylation site or epitope without separate validation. |
| IF/ICC Q&A: What should fluorescence show? | HPA reports vesicles as the enhanced main ICC-IF location, with images from A-431, U-251MG, U2OS and NIH 3T3 (HPA: subcellular). Expect a vesicular signal when assessing IF/ICC, while interpreting that application on its own guide page; the tissue IHC pattern describes paraffin-section chromogenic staining (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| A proposed positive control is faint or negative. | The sampled field may lack the specific high-staining cell population, or a low-staining population may have been selected (HPA: tissue IHC). | Confirm cell identity and use a section containing an HPA-listed high-staining glandular or endocrine population; then check the IHC-P staining and detection steps (HPA: tissue IHC; standard IHC practice). |
| Myocytes appear strongly positive. | HPA reports skeletal myocytes and smooth muscle cells as not detected; unexpected colour could arise from nonspecific antibody binding or endogenous enzyme activity in chromogenic detection (HPA: tissue IHC; standard IHC practice). | Check the no-primary control and cell identity. If an enzyme-based system is used, review its endogenous-activity control before interpreting the myocyte signal (standard IHC practice). |
| Colour is widespread and cell boundaries cannot be resolved. | Diffuse background can obscure the expected cytoplasmic and membranous distribution (HPA: tissue IHC; standard IHC practice). | Compare with a no-primary control and review blocking, washing and chromogen development; score only fields where individual cells and compartments remain distinguishable (standard IHC practice). |
| The signal appears exclusively nuclear. | Nuclear localisation can occur in some cells, but the reported tissue IHC profile is predominantly cytoplasmic and membranous (UniProt Q13492; HPA: tissue IHC). | Recheck the same slide for the expected cytoplasmic pattern, compare controls and avoid calling nuclear-only staining a confirmed PICALM pattern without further validation (standard IHC practice). |
| A strong signal forms a continuous edge around nearly every cell. | PICALM associates with coated pits and vesicles and lacks a transmembrane segment; an uninterrupted rim alone does not establish the reported compartment pattern (UniProt Q13492; HPA: tissue IHC). | Inspect higher magnification for cytoplasmic or vesicular detail, compare the no-primary control and judge staining within the relevant cell types (HPA: tissue IHC; standard IHC practice). |
| A positive tissue remains unstained after cell identity is confirmed. | The evidence supplied does not establish a PICALM-specific fixation or retrieval failure; absent staining can reflect a problem elsewhere in the IHC-P workflow (standard IHC practice). | Check that the antibody and detection reagents were applied correctly, that the control run developed, and that retrieval followed the catalog antibody's IHC-P instructions (standard IHC practice). |
Experiment evidence
IHC & IF images showing PICALM staining
A02053-1 · IHCIHC analysis of PICALM using anti-PICALM antibody (A02053-1). PICALM was detected in paraffin-embedded section of human Ovarian cancer tissu
A02053-1 · IHCIHC analysis of PICALM using anti-PICALM antibody (A02053-1). PICALM was detected in paraffin-embedded section of human Ovarian cancer tissu
A02053-1 · IFIF analysis of PICALM using anti-PICALM antibody (A02053-1). PICALM was detected in immunocytochemical section of A431 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000073921-PICALM/tissue/cerebral+cortex#imid_5329008
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000073921-PICALM/tissue/colon#imid_5328950
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000073921-PICALM/tissue/liver#imid_5328943
HPA · PancreasHuman Protein Atlas IHC, Pancreas. Source: https://www.proteinatlas.org/ENSG00000073921-PICALM/tissue/pancreas#imid_5328886
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019053; green = PICALM, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA019053; green = PICALM, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PICALM IHC & IF
🧪Run breast first: its glandular cells should stain (HPA: High in breast glandular cells). Use skeletal muscle as the negative tissue (HPA: Not detected in skeletal muscle myocytes); on the breast slide, cells without specific signal should show counterstain without brown DAB, but the supplied HPA data do not identify a particular breast cell type as negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Skeletal muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PICALM in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Vesicles (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls, plus a PICALM knockout sample processed alongside the test section (selected-SKU caption: rabbit primary antibody; standard IHC controls). Quench endogenous peroxidase and assess endogenous biotin background because the caption uses a streptavidin–biotin complex with DAB (selected-SKU caption: SABC/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the exact selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used in that IHC example, but retrieval dependence has not been established (selected-SKU caption: heat retrieval). ICC-IF images show vesicular localisation, while the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC; no breast-specific artefact is reported (HPA: vesicles enhanced; selected-SKU caption: paraffin-section IHC).
HPA tissue IHC evidence for PICALM
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 → |
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
Section 3
Advanced PICALM IHC Tips
Use the documented paraffin-section workflow as a starting point, then assess staining by compartment, cell type and matched controls (datasheet A02053-1; HPA tissue IHC).
What retrieval should I try first when PICALM staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A02053-1). The documented workflow then uses 10% goat-serum blocking and 1 μg/ml primary antibody overnight at 4°C (datasheet A02053-1). Because the retrieval heating time is unreported, document heating and cooling conditions and compare adjacent sections while keeping antibody concentration and detection constant (datasheet A02053-1; standard IHC practice). If signal remains weak, test another retrieval condition as a fallback and judge signal against tissue preservation, background and a primary-antibody omission control (standard IHC practice).
Does PICALM require a particular fixation time for paraffin IHC?
PICALM-specific fixation sensitivity is unknown: the selected paraffin-section caption does not report a fixative or fixation time (datasheet A02053-1). Do not treat its successful staining as evidence for a particular fixation schedule, or infer fixation tolerance from PICALM topology or phosphorylation sites (datasheet A02053-1; UniProt Q13492 topology and modified residues). For troubleshooting, compare sections with documented fixation histories under the same EDTA pH 8.0 retrieval and detection conditions (datasheet A02053-1; standard IHC practice). Assess morphology and background alongside signal, using matched tissue controls before changing retrieval or antibody concentration (standard IHC practice).
Which staining compartments should count as plausible PICALM signal?
Prioritise cytoplasmic and membranous staining in paraffin sections, the predominant tissue pattern reported for PICALM (HPA tissue IHC). Vesicular staining is also plausible: PICALM localises to clathrin-coated pits, coated vesicles and the Golgi area, and ICC/IF reports an enhanced vesicular location (UniProt Q13492 subcellular; HPA subcellular). Nuclear staining requires closer scrutiny because PICALM can enter the nucleus in some cells through interaction with PIMREG, but this is context dependent (UniProt Q13492 subcellular). Compare compartment patterns with adjacent morphology and omission controls; uniform nuclear DAB across unrelated cells should prompt a background check (standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent PICALM IHC staining?
PICALM has 5 reported isoforms, but the selected IHC caption does not map the antibody epitope or establish which isoforms it detects (UniProt Q13492 isoforms; datasheet A02053-1). Its ENTH domain spans residues 14–145, and reported phosphorylation includes residues 16, 20, 303 and 315; none establishes this antibody's binding site (UniProt Q13492 domains and modified residues). Obtain epitope or immunogen information before attributing different tissue patterns to an isoform or modification (standard IHC interpretation). Meanwhile, compare matched sections using the documented EDTA pH 8.0 retrieval and consistent detection settings (datasheet A02053-1; standard IHC practice).
How should I add IF to a PICALM paraffin-IHC study?
Treat IF as a separate assay and multiplex PICALM with a marker identifying the expected cell population; glandular cells stain highly in several reported tissues (HPA tissue IHC). Choose a fluorophore away from the tissue's strongest autofluorescence, assess single-colour controls, and use spectral separation when channels overlap (standard IF practice). PICALM has no transmembrane segment and associates with cytoplasmic membrane structures, but the catalog antibody's epitope side is unreported; select and titrate permeabilisation only after establishing epitope accessibility (UniProt Q13492 topology and subcellular; datasheet A02053-1; standard IF practice). Compare vesicular localisation with the reported ICC/IF pattern, without treating the chromogenic paraffin caption as IF validation (HPA subcellular; datasheet A02053-1).
How can I distinguish PICALM staining from chromogenic background?
The documented paraffin workflow uses a biotinylated secondary, streptavidin–biotin complex and DAB, so assess background from those detection steps with a primary-antibody omission control (datasheet A02053-1; standard IHC practice). Include an appropriate peroxidase block before DAB development and check whether residual enzyme activity produces colour without primary antibody (standard chromogenic IHC practice). If widespread colour persists, assess endogenous biotin interference or compare a nonbiotin detection system while holding tissue preparation constant (standard IHC practice). Interpret the result against PICALM's reported cytoplasmic and membranous tissue pattern, rather than accepting diffuse colour alone as target staining (HPA tissue IHC).
How should I score PICALM staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic or membranous compartment before scoring, reflecting the reported tissue staining pattern (HPA tissue IHC). For intensity, use an H-score from 0–300 by summing the percentage of cells at each intensity multiplied by its 0–3 score; also report percentage positive cells when useful (standard IHC scoring practice). For spatial analysis, count positive cells per mm² of viable tissue and normalise to the same cell population or viable area across sections (standard IHC quantification practice). Keep staining batch, threshold and sampling rules consistent, and record background separately from biological signal (standard IHC practice).
When is an unexpected PICALM IHC pattern likely artefactual?
A credible result should fit tissue morphology and the reported cytoplasmic or membranous pattern, with vesicle-associated signal also biologically plausible (HPA tissue IHC; UniProt Q13492 subcellular). Compare strongly stained cells with the expected population: glandular or endocrine cells are reported high in several tissues, whereas skeletal myocytes and smooth muscle cells are reported undetected (HPA tissue IHC). Treat edge-restricted colour, staining confined to necrotic regions, or colour in omission controls as potential processing or detection artefacts (standard IHC practice). Scrutinise isolated nuclear signal separately because nuclear localisation is possible in some cells, while endogenous enzyme activity can confound DAB interpretation (UniProt Q13492 subcellular; standard IHC practice).
Boster reagents
Best PICALM / Phosphatidylinositol-binding clathrin assembly protein IHC Antibodies
A02053-1 has IHC data from a human ovarian cancer paraffin section and IF data from A431 cells; its listed reactivity includes human, mouse and rat (catalog image captions; catalog reactivity).
Real IHC data
Anti-PICALM ® Antibody
Cat # A02053-1
A02053-1 will render with IHC from a human ovarian cancer paraffin section and IF from A431 cells (A02053-1 image captions). M02053 will render as a human-reactive IHC and IF/ICC antibody, with no image-based sample validation in the payload (M02053 applications, reactivity and image captions).
Which to pick: For tissue IHC, choose A02053-1 when a demonstrated paraffin-section workflow matters: its IHC caption reports EDTA pH 8 retrieval and 1 μg/ml primary antibody, but does not report the fixative (A02053-1 IHC caption). For IF/ICC, A02053-1 has an A431-cell IF image; M02053 is a monoclonal alternative listed for human IF/ICC and IHC, without a sample image in this payload (A02053-1 IF caption; M02053 catalog applications, clone and image captions). For mouse or rat samples, A02053-1 is the listed cross-species choice, although its supplied IHC and IF images show human samples (A02053-1 catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q13492 (PICAL_HUMAN, Phosphatidylinositol-binding clathrin assembly protein).
- Human Protein Atlas. PICALM tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. PICALM subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. PICALM antibody validation summary (2 antibodies).
- A single codon insertion in PICALM is associated with development of familial subvalvular aortic stenosis in Newfoundland dogs. Human genetics 2014 — PMC4148152.
- Genetics of PICALM expression and Alzheimer's disease. PloS one 2014 — PMC3949918.
- PICALM exerts a role in promoting CRC progression through ERK/MAPK signaling pathway. Cancer cell international 2022 — PMC9063212.
- Central role for PICALM in amyloid-β blood-brain barrier transcytosis and clearance. Nature neuroscience 2015 — PMC4482781.
- PubMed PMID:8643484 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16554811 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PICALM (Phosphatidylinositol-binding clathrin assembly protein). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02053-1). Validated IHC antibody: A02053-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Skeletal muscle. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human Ovarian cancer tissue; fixative not specified (datasheet A02053-1); 2. Fixation: Image fixative and duration unreported (datasheet A02053-1); 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 A02053-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02053-1); 8. Primary antibody: Rabbit anti-PICALM, 0.5-1μg/ml (datasheet A02053-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02053-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium concordance (HPA tissue IHC); Isoform / epitope: 5 isoforms; verify antibody epitope coverage (UniProt); Regulation: Broad expression across examined tissues (UniProt). Evidence: UniProt Q13492, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A02053-1, PMC4148152, PMC3949918. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PICALM/immunohistochemistry-ihc-guide.