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LRRK2 / Leucine-rich repeat serine/threonine-protein kinase 2 · IHC design guide
Design Immunohistochemistry for LRRK2
Plan LRRK2 paraffin IHC with the catalog antibody at 1:25 (datasheet M00221-3). Use strongly stained kidney tubules as a reference and assess cytoplasmic staining in the relevant cell types (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LRRK2 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 staining in several cell types (HPA tissue IHC) | |
| Staining pattern | High in renal tubules; medium in alveolar cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet M00221-3) | |
| Positive control | Kidney+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formaldehyde-fixed paraffin sections (selected-SKU IHC image M00221-3) | |
| Caveat | Brain expression may yield low neuronal staining (UniProt; HPA tissue IHC) | |
| Regulation | Lysosomal stress recruits LRRK2 (UniProt) | |
| Isoform / epitope | No annotated isoforms or transmembrane segment (UniProt) |
Section 1
Recommended LRRK2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses citrate pH 6 retrieval (datasheet: M00221-3). The published IHC protocols below cover rodent brain and tissue sections (PMC4076169; PMC5289148; PMC7812009).
| Sample | Formaldehyde-fixed, paraffin-embedded Human brain tissue (datasheet M00221-3) |
| Fixation | Image formalin-fixed; duration unreported (datasheet M00221-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet M00221-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-LRRK2, 1:25 (datasheet M00221-3) |
| Primary incubation | 1 hours at 37°C (datasheet M00221-3) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LRRK2-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several cell types, including pneumocytes and cells in renal tubules. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval for the catalog antibody (datasheet: M00221-3). For rodent brain, stain freshly sectioned tissue when possible (PMC4076169).
Section 2
What Is the Expected LRRK2 Staining Pattern?
LRRK2 is expected mainly in cytoplasmic and vesicle-associated compartments; it has no transmembrane segment (UniProt Q5S007 topology and subcellular location). In paraffin-section IHC, look first at renal tubular cells, reported High, and lung alveolar cells, reported Medium (HPA tissue IHC). HPA calls its tissue IHC reliability Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Distinct cytoplasmic staining in renal tubular cells, with weaker staining in lung alveolar cells. | This matches the reported High tubular and Medium alveolar staining levels (HPA tissue IHC). Compare cells within each section: these levels describe HPA observations, not intensity thresholds for every run (HPA tissue IHC; general IHC practice). |
| Strong nuclear-only staining or a uniform nuclear outline dominates the positive cells. | Recheck compartment assignment and controls: UniProt places LRRK2 chiefly in cytoplasmic and vesicle-associated locations (UniProt Q5S007 subcellular location). HPA ICC-IF lists nucleoplasm and nuclear membrane only as uncertain additional locations, so nuclear-only IHC does not establish the expected pattern (HPA subcellular ICC-IF). |
| Prominent staining appears in cells reported as undetected, such as adipocytes in adipose tissue. | Consider antibody cross-reactivity or endogenous chromogenic detection activity (HPA tissue IHC: adipocytes Not detected; general IHC practice). Check cell identity and a matched negative control before assigning that signal to LRRK2 (general IHC practice). |
| Colour covers the section diffusely, obscuring cell borders and cytoplasmic detail. | The pattern cannot be scored against HPA's cell-specific IHC observations (HPA tissue IHC; general IHC practice). Inadequate blocking, insufficient washing, or excess detection reagent can produce diffuse background; inspect the no-primary control to locate the source (general IHC practice). |
| Renal tubular cells show no discernible signal in an otherwise readable section. | HPA reports High staining in renal tubular cells, making them a useful reference for a missed signal (HPA tissue IHC). Check the IHC-validated antibody, detection reagents, and retrieval conditions with controls before interpreting an unstained specimen as LRRK2-negative (general IHC practice). |
💡Expected LRRK2 appearanceCall a result positive when renal tubular cell cytoplasm is clearly stained and lung alveolar cells can show less intense cytoplasmic staining (HPA tissue IHC: High and Medium); strong nuclear-only or widespread cell-independent colour warrants control-based review (UniProt Q5S007 subcellular location; general IHC practice).
| Tissue and cell selection | Use renal tubular cells as the stronger reference and lung alveolar cells as a second observed site (HPA tissue IHC: High; Medium). HPA reports several low-level sites, so a faint result there needs careful comparison with controls (HPA tissue IHC; general IHC practice). |
| Subcellular interpretation | LRRK2 is associated with vesicles, Golgi, endosomes and lysosomes, among other cytoplasmic sites (UniProt Q5S007 subcellular location). Its lack of a transmembrane segment does not make crisp cell-surface staining the expected IHC result (UniProt Q5S007 topology; general IHC practice). |
| Antibody validation | HPA lists HPA014293 and CAB037160 as IHC Approved; HPA014293 is also ICC Supported (HPA antibodies). These labels support the reported uses but do not independently verify an unexpected cell or compartment in a new specimen (HPA antibodies; general IHC practice). |
| IF/ICC comparison | On its separate IF/ICC guide, the relevant comparison is vesicular localisation: HPA marks vesicles Supported, while basal body and listed nuclear locations are Uncertain (HPA subcellular ICC-IF). Treat those uncertain locations as prompts for validation, not IHC acceptance criteria (HPA subcellular ICC-IF; general IHC practice). |
| Antigen retrieval and detection | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Situation | Likely cause | Next action |
|---|---|---|
| Renal tubules are negative while the section and counterstain look intact. | A failed primary, retrieval, or detection step is possible; HPA reports High tubular staining (general IHC practice; HPA tissue IHC). | Run a known-positive kidney section alongside the specimen; verify the catalog antibody's IHC instructions and the detection controls before scoring absence (general IHC practice). |
| Lung alveolar cells stain weakly, with a clear kidney positive control. | This may fit the reported Medium alveolar versus High tubular levels (HPA tissue IHC). | Compare the signal with nearby background and the no-primary control; avoid calling weak alveolar staining a failure solely because it is less intense than kidney (HPA tissue IHC; general IHC practice). |
| A tissue reported as Not detected shows broad staining in many cell types. | Nonspecific antibody binding or endogenous chromogenic activity may account for the mismatch (HPA tissue IHC; general IHC practice). | Check the exact cell type against HPA, examine a no-primary control, and use the appropriate endogenous-enzyme block for the chosen chromogen system (HPA tissue IHC; general IHC practice). |
| Signal is mainly nuclear or outlines the plasma membrane. | That distribution conflicts with the expected cytoplasmic and vesicle-associated pattern; HPA's additional nuclear ICC-IF locations remain Uncertain (UniProt Q5S007 subcellular location; HPA subcellular ICC-IF). | Review morphology and control sections, then confirm the pattern with another independently validated antibody if available (general IHC practice). |
| Diffuse colour hides individual positive cells. | Background from blocking, washing, primary concentration, or detection can prevent cell-specific interpretation (general IHC practice). | Compare with a no-primary control; adjust one general IHC variable at a time using the catalog antibody's IHC instructions (general IHC practice). |
| Cortical glia are negative despite reported LRRK2 expression in brain. | The records refer to different cells: HPA reports cerebral-cortex glial cells Not detected, while UniProt reports cortical pyramidal neurons expressing LRRK2 (HPA tissue IHC; UniProt Q5S007 tissue specificity). | Identify neurons and glia separately before scoring; use the observed kidney tubular pattern as a positive IHC reference (HPA tissue IHC; UniProt Q5S007 tissue specificity). |
Experiment evidence
IHC & IF images showing LRRK2 staining
M00221-3 · IHCM00221-3 staining LRRK2 in Human brain tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections).
M00221-2 · IHCImmunohistochemical analysis of paraffin-embedded human thyroid, using LRRK2 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000188906-LRRK2/tissue/cerebral+cortex#imid_3683848
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000188906-LRRK2/tissue/lung#imid_3683719
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000188906-LRRK2/tissue/colon#imid_3683758
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000188906-LRRK2/tissue/liver#imid_3683767
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA014293; green = LRRK2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA014293; green = LRRK2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LRRK2 IHC & IF
🧪Run kidney first: its tubular cells should stain strongly (HPA: High in kidney cells in tubules). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, treat cells outside the tubules as candidate internal negatives only if they show background-level staining, since their LRRK2 status is unspecified (HPA: kidney row identifies tubular cells only).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LRRK2 in A-549, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Vesicles (supported), Basal body (uncertain) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a primary antibody host-species-, class-, and clonality-matched isotype control, and LRRK2 knockout material as a biological negative where available (standard IHC practice). Check endogenous peroxidase activity and block endogenous biotin in kidney sections when using chromogenic biotin-based detection (standard IHC practice; selected IHC caption: biotinylated secondary antibody).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU tissue-IHC caption does not establish a fixative despite its processing narrative (selected IHC caption: fixative unreported). Heat-mediated citrate retrieval at pH 6 was used in the reported paraffin-section example, but retrieval dependency has not been established (selected IHC caption: citrate buffer, pH 6). Neither an advantage for frozen sections nor an easier IF workflow is established by the supplied evidence; for kidney IHC, assess endogenous biotin and peroxidase background (standard IHC practice).
HPA tissue IHC evidence for LRRK2
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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LRRK2 IHC Tips
Troubleshoot LRRK2 staining in paraffin sections by checking retrieval, cellular pattern and controls before comparing chromogenic signal across samples.
How should I adjust retrieval when LRRK2 staining is weak or uneven?
Start with heat-mediated citrate retrieval at pH 6 for paraffin sections (datasheet M00221-3). The selected brain IHC image used that retrieval before a 1:25 primary incubation for 1 hour at 37°C (caption M00221-3). If signal is weak, check deparaffinisation, heating uniformity and section adherence, then titrate retrieval time while holding antibody concentration and detection constant (standard IHC practice). Compare each condition with the same positive control and a primary-antibody omission control; stronger staining alone does not establish specificity (standard IHC practice). Record whether improved signal remains cytoplasmic and cell associated, consistent with reported LRRK2 tissue staining (HPA tissue IHC).
Could fixation explain inconsistent LRRK2 staining between paraffin blocks?
LRRK2-specific sensitivity to fixation is unknown from the supplied evidence, so treat differences between blocks as a question to test rather than an established protein effect. Compare sections with documented fixation and processing histories, using the same pH 6 citrate retrieval and staining run for each block (datasheet M00221-3; standard IHC practice). Check morphology and counterstaining alongside the LRRK2 signal, since poor preservation can make cell assignment unreliable (standard IHC practice). Keep a reference section in every run, and avoid interpreting a weaker block as lower biological expression until technical variation has been assessed (standard IHC practice).
Which subcellular pattern is plausible for LRRK2 in chromogenic tissue sections?
Look first for cell-associated cytoplasmic staining, the pattern reported across several tissue cell types (HPA tissue IHC). LRRK2 is associated with vesicles, endosomes, lysosomes, Golgi and other intracellular structures, so granular or uneven cytoplasmic signal can be plausible (UniProt Q5S007 subcellular location). A paraffin section with chromogenic detection may not resolve those structures individually; assess the whole cell and its neighbours before assigning an organelle (standard IHC practice). Treat predominantly nuclear staining cautiously: nucleoplasmic and nuclear-membrane locations are listed as uncertain in cellular imaging data (HPA subcellular). Compare the pattern with a primary-antibody omission control and a consistently stained reference section (standard IHC practice).
How do epitope location and phosphorylation affect LRRK2 IHC interpretation?
The supplied protein record lists one 1–2527 chain and 0 isoforms, with no transmembrane segment (UniProt Q5S007 sequence and topology). It also lists phosphorylation at Ser-910, Ser-935, Ser-955, Ser-973 and Ser-1292, among other modified residues (UniProt Q5S007 modified residues). Check the antibody’s stated immunogen and whether it recognises total protein or a modification-specific epitope before comparing staining across conditions (standard IHC practice). If the epitope is undisclosed, do not attribute a staining difference to a particular domain or phosphorylation site; use an independent antibody with a documented epitope when that distinction matters (standard IHC practice).
How can I check an apparent LRRK2 signal with multiplex immunofluorescence?
On the separate IF/ICC workflow, pair LRRK2 with a validated marker for the cell population being assessed; tubular cells and alveolar cells are reported staining populations in tissue (HPA tissue IHC). Choose spectrally separated fluorophores and inspect unstained tissue in each channel to identify autofluorescence before interpreting overlap (standard IF practice). LRRK2 has no transmembrane segment and is reported at intracellular vesicles and membranes, so select permeabilisation according to the antibody’s documented epitope accessibility and the structures under study (UniProt Q5S007 topology and subcellular location; standard IF practice). Validate fixation, permeabilisation and channel controls in IF/ICC independently; the IHC image does not establish IF/ICC performance (caption M00221-3; standard IF practice).
What should I check when LRRK2 chromogen appears throughout the section?
First compare a primary-antibody omission control with the stained slide to separate detection-system background from antibody-dependent signal (standard IHC practice). The selected image used 3% BSA for 0.5 hour and an undiluted biotinylated secondary antibody (caption M00221-3). With a biotin-based system, check endogenous biotin and include an appropriate block if it contributes signal; also block endogenous peroxidase before chromogenic development (standard IHC practice). Titrate primary antibody and DAB development against a reference section while preserving cell morphology (standard IHC practice). Diffuse staining outside identifiable cells should not be called LRRK2 solely because its intensity increases with development time (standard IHC practice).
How should I score LRRK2 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring: HPA reports high staining in kidney tubular cells and medium staining in lung alveolar cells (HPA tissue IHC). For chromogenic sections, record the percentage of positive cells and an intensity-based H-score, or count positive cells per mm² when cell density is the main endpoint (standard IHC practice). Normalise cell counts to the annotated viable tissue area, and compare intensity scores only among sections stained and imaged under matched conditions (standard IHC practice). Exclude folds, edges and necrotic areas by a prespecified rule, then report the number of sections and sampled regions so apparent differences can be checked (standard IHC practice).
How can I distinguish genuine LRRK2 staining from tissue artefact?
A credible result has a reproducible cell-associated cytoplasmic pattern and appears in an expected cell population, such as kidney tubular cells (HPA tissue IHC). LRRK2 has reported vesicular and other intracellular locations, whereas nuclear locations in cellular imaging carry uncertain support (UniProt Q5S007 subcellular location; HPA subcellular). Inspect edge-heavy staining, necrosis and pigment separately, and compare them with the primary-antibody omission and endogenous-peroxidase controls (standard IHC practice). Do not infer LRRK2 kinase activity from total-protein chromogenic staining: the record describes phosphorylation-dependent regulation, while this tissue stain establishes neither modification state nor enzymatic activity (UniProt Q5S007 function and modified residues; standard IHC interpretation).
Boster reagents
Best LRRK2 / Leucine-rich repeat serine/threonine-protein kinase 2 IHC Antibodies
The catalog includes IHC images from human brain and thyroid paraffin sections; IF/ICC applications are listed for human, mouse, and rat, but no IF image is supplied (catalog applications and reactivity; IHC image captions).
Real IHC data
Anti-LRRK2 Antibody (C-term)
Cat # M00221-3
Real IHC data
Anti-LRRK2 Rabbit Monoclonal Antibody
Cat # M00221-2
M00221-3 has an IHC image from formaldehyde-fixed human brain paraffin sections (M00221-3 IHC image caption). M00221-2 has an IHC image from human thyroid paraffin sections; its fixative is unreported, and IF/ICC is listed without an IF image (M00221-2 IHC image caption; catalog applications).
Which to pick: For tissue IHC with a documented processing example, choose rabbit polyclonal M00221-3: its human brain caption reports formaldehyde fixation, citrate retrieval at pH 6, and primary antibody at 1:25 (catalog clonality; M00221-3 IHC image caption). For IF/ICC or work across human, mouse, and rat, choose rabbit monoclonal M00221-2 based on its listed applications and reactivity; its own IHC image documents human thyroid paraffin sections, with the fixative unreported (catalog applications, reactivity, and clonality; M00221-2 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5S007 (LRRK2_HUMAN, Leucine-rich repeat serine/threonine-protein kinase 2).
- Human Protein Atlas. LRRK2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LRRK2 subcellular location (ICC-IF): Mainly localized to vesicles and basal body. In addition localized to the nucleoplasm, nuclear membrane and cytosol..
- Human Protein Atlas. LRRK2 antibody validation summary (2 antibodies).
- Differential LRRK2 expression in the cortex, striatum, and substantia nigra in transgenic and nontransgenic rodents. The Journal of comparative neurology 2014 — PMC4076169.
- Behavioral, neurochemical, and pathologic alterations in bacterial artificial chromosome transgenic G2019S leucine-rich repeated kinase 2 rats. Neurobiology of aging 2015 — PMC5289148.
- LRRK2 interactions with microtubules are independent of LRRK2-mediated Rab phosphorylation. EMBO reports 2025 — PMC12238589.
- TM4SF4 and LRRK2 Are Potential Therapeutic Targets in Lung and Breast Cancers through Outlier Analysis. Cancer research and treatment 2021 — PMC7812009.
- PubMed PMID:15541309 — UniProt-cited evidence.
- PubMed PMID:16541075 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LRRK2 (Leucine-rich repeat serine/threonine-protein kinase 2). Expected localisation Cytoplasmic staining in several cell types (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet M00221-3). Validated IHC antibody: M00221-3 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded Human brain tissue (datasheet M00221-3); 2. Fixation: Image formalin-fixed; duration unreported (datasheet M00221-3); 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: Citrate pH 6 (datasheet M00221-3); 20 min, 95–100 °C (standard); 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: Rabbit anti-LRRK2, 1:25 (datasheet M00221-3); 9. Primary incubation: 1 hours at 37°C (datasheet M00221-3). Watch for: Fixation: Formaldehyde-fixed paraffin sections (selected-SKU IHC image M00221-3); Caveat: Brain expression may yield low neuronal staining (UniProt; HPA tissue IHC); Isoform / epitope: No annotated isoforms or transmembrane segment (UniProt); Regulation: Lysosomal stress recruits LRRK2 (UniProt). Evidence: UniProt Q5S007, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to vesicles and basal body. In addition localized…, Datasheet M00221-3, PMC4076169, PMC5289148. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LRRK2/immunohistochemistry-ihc-guide.