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LMTK3 / Serine/threonine-protein kinase LMTK3 · IHC design guide
Design Immunohistochemistry for LMTK3
Plan chromogenic paraffin IHC for LMTK3 using neuronal cytoplasmic and axonal staining as the tissue reference (HPA tissue IHC). Start the catalog antibody at 0.5–1 μg/mL (datasheet A08432-2) and score neuronal cell bodies and axons (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LMTK3 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 | Neuronal cytoplasm and axons (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and axonal staining in neuronal cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A08432-2) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08432-2) | |
| Caveat | Scoring cell bodies alone can miss axonal staining (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; signal peptide 1–20 is cleaved, so check epitope position (UniProt) |
Section 1
Recommended LMTK3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A08432-2) is accompanied by published gastric and thyroid cancer protocols (PMC3810398; PMC5364963).
| Sample | Paraffin-embedded mouse brain tissues; fixative not specified (datasheet A08432-2) |
| Fixation | Image fixative and duration unreported (datasheet A08432-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A08432-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08432-2) |
| Primary antibody | Rabbit anti-LMTK3, 0.5-1μg/ml (datasheet A08432-2) |
| Primary incubation | Overnight at 4 °C (datasheet A08432-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A08432-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LMTK3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and axonal expression in neuronal cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet: A08432-2). The thyroid study also used EDTA at pH 8 after citrate (PMC5364963).
Section 2
What Is the Expected LMTK3 Staining Pattern?
LMTK3 is a membrane protein with a transmembrane segment at residues 40–60 and is also annotated at the Golgi and in neuronal projections (UniProt Q96Q04). In tissue IHC, expect cytoplasmic and axonal staining in neurons, with high staining reported in several brain regions and adrenal glandular cells (HPA: tissue IHC; reliability Enhanced). Read the cellular pattern together with tissue identity; membrane topology alone does not define the appearance of a chromogenic section (UniProt Q96Q04 topology).
| Neuronal cytoplasm and axonal processes stain in cerebral cortex, hippocampus or caudate; Purkinje cells stain in cerebellum (HPA: High in these cells). | This fits the reported tissue pattern: cytoplasmic and axonal expression in neuronal cells (HPA: tissue IHC). Assess the relevant cell population and its processes, rather than treating every stained structure in the section as an equivalent positive result (HPA: tissue IHC; general IHC practice). |
| Adrenal glandular cells show strong staining, while smooth muscle cells or small-intestinal enterocyte microvilli show less staining (HPA: High in adrenal; Medium in smooth muscle and microvilli). | These intensities are compatible with the listed cell-specific observations (HPA: tissue IHC). The microvillus entry describes a particular compartment, so assess whether staining follows that structure before calling the surrounding enterocyte cytoplasm positive (HPA: Medium in enterocyte microvilli). |
| A brain section shows predominantly isolated nuclear staining, without the reported neuronal cytoplasmic or axonal pattern (HPA: tissue IHC). | That is an atypical IHC result requiring review of specificity and section controls (HPA: cytoplasmic and axonal neuronal profile; general IHC practice). HPA's uncertain nucleoplasmic ICC-IF annotation does not establish nuclear staining as the expected tissue IHC pattern (HPA: subcellular ICC-IF, uncertain). |
| Adipocytes or bone-marrow hematopoietic cells stain prominently (HPA: Not detected in those cell populations). | Consider off-target binding or endogenous chromogenic detection activity before interpreting these cells as LMTK3 positive (HPA: tissue IHC; general IHC practice). Compare an appropriate reagent control with the tissue pattern; a negative cell population in one atlas entry is a reference point, not proof that every specimen must be blank (HPA: tissue IHC). |
| Color appears broadly across cells and extracellular areas, with little separation between neuronal processes and the surrounding section (HPA: neuronal profile). | Treat this as background until controls show a cell-specific signal (general IHC practice). Review blocking, washes, detection reagents and counterstain as general workflow variables; diffuse color alone cannot be assigned to LMTK3 or used to infer a target-specific fixation effect (general IHC practice). |
💡Expected LMTK3 appearanceA convincing positive is cell-resolved cytoplasmic and axonal staining in neurons, with High staining reported in several brain regions; widespread color in HPA-listed negative cell populations or isolated nuclear-only IHC staining warrants a specificity check (HPA: tissue IHC; HPA: subcellular ICC-IF, uncertain).
| Cell and compartment chosen for scoring (HPA: tissue IHC) | HPA reports High neuronal staining in caudate, cortex and hippocampus and High Purkinje-cell staining in cerebellum, alongside cytoplasmic and axonal localisation (HPA: tissue IHC). Score the named cells and their reported compartments rather than averaging the whole section (general IHC practice). |
| Antibody evidence for tissue IHC (HPA: antibody validation) | HPA lists HPA077070 as IHC Enhanced and HPA040203 as ICC Approved, with no IHC status listed for HPA040203 (HPA: antibodies). Enhanced describes consistency between staining and RNA data here; it supports the tissue pattern but does not guarantee that another antibody or specimen will reproduce it (HPA: tissue IHC reliability). |
| Membrane topology and cellular distribution (UniProt Q96Q04) | The transmembrane segment at residues 40–60 and annotations for membrane, Golgi membrane, axon and dendrite provide context for nonnuclear structures (UniProt Q96Q04). An antibody epitope is not supplied, so topology cannot predict which processing or retrieval condition exposes it (UniProt Q96Q04 topology; epitope unspecified in supplied record). |
| IF/ICC Q&A: Should nucleoplasmic IF set the IHC scoring rule? (HPA: subcellular ICC-IF) | No. HPA calls nucleoplasm the main ICC-IF location and cell junctions an additional location, both uncertain (HPA: subcellular ICC-IF). For paraffin-section IHC, use the separately reported cytoplasmic and axonal neuronal pattern as the tissue reference (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in a known-positive brain region (HPA: High in cortex, hippocampus and caudate). | The run may have failed at antibody binding, detection or another general IHC step; the supplied sources do not identify LMTK3-specific fixation sensitivity (general IHC practice; HPA: tissue IHC). | Check the positive-control section and reagent controls, then review retrieval and detection settings under the antibody's documented IHC procedure (general IHC practice). |
| Only nuclei stain in a neuronal section (HPA: cytoplasmic and axonal tissue profile). | The compartment conflicts with the reported IHC pattern; the uncertain ICC-IF nucleoplasmic finding cannot resolve tissue-IHC specificity (HPA: tissue IHC; HPA: subcellular ICC-IF). | Inspect cell boundaries and counterstain, compare controls, and avoid scoring nuclear-only color as the expected IHC result (general IHC practice; HPA: tissue IHC). |
| Negative-reference adipocytes or marrow hematopoietic cells are strongly colored (HPA: Not detected). | Off-target signal or endogenous detection activity is plausible, though the stain alone cannot distinguish them (HPA: tissue IHC; general IHC practice). | Compare omission or detection controls and assess whether the expected positive cell pattern remains distinct (general IHC practice; HPA: tissue IHC). |
| Signal is diffuse and tissue detail is obscured. | Background from incomplete blocking, washing or chromogen development may obscure a specific pattern (general IHC practice). | Review the general blocking, wash and detection steps, then reassess whether neuronal cytoplasm and axons remain distinguishable (general IHC practice; HPA: tissue IHC). |
| Adrenal gland stains, but a small-intestinal section looks weaker (HPA: High versus Medium). | Different listed cells and levels can account for this contrast; HPA specifies enterocyte microvilli rather than all enterocyte compartments (HPA: tissue IHC). | Identify glandular cells and the microvillus border before comparing intensity, and record cell-specific scores (HPA: tissue IHC; general IHC practice). |
| A reagent qualified for ICC gives an unexpected paraffin-section result (HPA: antibody validation). | HPA lists ICC Approved for HPA040203 without an IHC status, while HPA077070 carries IHC Enhanced status (HPA: antibodies). | Check which antibody was used and evaluate paraffin-section staining against the IHC-supported reagent and tissue pattern (HPA: antibodies; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing LMTK3 staining
A08432-2 · IHCIHC analysis of LMTK3 using anti-LMTK3 antibody (A08432-2). LMTK3 was detected in paraffin-embedded section of mouse brain tissues. Heat med
A08432-2 · IHCIHC analysis of LMTK3 using anti-LMTK3 antibody (A08432-2). LMTK3 was detected in paraffin-embedded section of mouse brain tissues. Heat med
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142235-LMTK3/tissue/cerebral+cortex#imid_1224
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142235-LMTK3/tissue/cerebral+cortex#imid_20432363
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142235-LMTK3/tissue/colon#imid_20432273
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142235-LMTK3/tissue/liver#imid_20432282
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040203; green = LMTK3, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA040203; green = LMTK3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LMTK3 IHC & IF
🧪Run cerebellum first and confirm staining in Purkinje cells (HPA: High in Purkinje cells). Run adipose tissue as the negative comparison (HPA: Not detected in adipocytes); on the cerebellum slide, assess neighboring cells without specific staining as internal background controls, without assuming that every other cell type is negative (standard IHC control practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LMTK3 in A-431, MCF-7, U2OS, with annotated localisation: Nucleoplasm (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a host- and clonality-matched isotype control (caption: rabbit primary; clonality unreported); and LMTK3-knockout tissue as a biological negative if available (standard IHC control practice). Check endogenous peroxidase and biotin background in cerebellum because the captioned IHC detection uses a biotinylated secondary, streptavidin–biotin complex, and DAB (caption: A08432-2 detection); assess tissue autofluorescence if using IF (standard IF control practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A08432-2 paraffin-section caption does not state a fixative (caption: fixative not stated). The caption uses heat retrieval in citrate buffer at pH 6 for 20 minutes, but does not establish whether LMTK3 staining requires retrieval (caption: A08432-2). The supplied evidence does not establish whether frozen sections or IF are easier; for brain IF, check autofluorescence against a no-primary control (standard IF practice).
HPA tissue IHC evidence for LMTK3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LMTK3 IHC Tips
Troubleshoot LMTK3 staining in paraffin sections by checking retrieval, cell type and compartment before comparing chromogenic signal across samples.
How should I retrieve LMTK3 in paraffin sections when staining is weak?
Start with heat-mediated citrate pH 6 antigen retrieval for 20 minutes in paraffin sections (datasheet A08432-2). The selected IHC image used this treatment before overnight primary antibody incubation at 4°C, so it is the documented starting condition for this antibody (datasheet A08432-2). If signal remains weak, compare shorter and longer heating times on matched sections while holding antibody concentration and detection constant (standard IHC practice). Watch for tissue lifting or damaged morphology, and judge improvement in neuronal cell bodies and axons against a matched negative control (HPA: cytoplasmic and axonal neuronal expression; standard IHC practice).
Could fixation explain variable LMTK3 staining between paraffin blocks?
The selected paraffin-section caption does not state a fixative, so LMTK3-specific fixation sensitivity is unknown for this antibody (datasheet A08432-2). Record each block’s fixative and fixation duration before comparing staining, because processing differences can change antigen accessibility in routine IHC (standard IHC practice). Test blocks with comparable processing together using citrate pH 6 retrieval for 20 minutes and the same detection run (datasheet A08432-2; standard IHC practice). If results differ, vary retrieval on adjacent sections and assess morphology as well as signal; neither the tissue expression pattern nor LMTK3 topology establishes a fixation-specific effect (HPA: tissue IHC; UniProt Q96Q04 topology).
Which compartments should show credible LMTK3 staining in brain IHC?
Prioritize cytoplasmic and axonal staining in neuronal cells when reading brain sections (HPA: cytoplasmic and axonal neuronal expression). LMTK3 is also annotated at membranes, axons, dendrites and Golgi membranes, with puncta in cell projections, so a structured projection-associated pattern is plausible (UniProt Q96Q04 localisation). The selected antibody was demonstrated on a paraffin-embedded mouse brain section after 20 minutes of citrate pH 6 retrieval (datasheet A08432-2). Diffuse nuclear staining alone should prompt a specificity check: the HPA nucleoplasm assignment comes from ICC/IF and is marked uncertain, whereas its tissue IHC profile emphasizes neuronal cytoplasm and axons (HPA: subcellular; HPA: tissue IHC).
How does LMTK3 topology affect epitope accessibility in IHC?
LMTK3 has one annotated transmembrane segment at residues 40–60 and a kinase domain at 133–411 (UniProt Q96Q04 topology and domains). The supplied evidence does not identify this antibody’s epitope, so the sequence annotation cannot establish which membrane side its binding site occupies (datasheet A08432-2; UniProt Q96Q04 topology). Two annotated glycosylation sites, at 1081 and 1405, and several modified residues could matter only if an independently mapped epitope overlaps them (UniProt Q96Q04 modifications). No alternative isoforms are listed; compare staining with an independently validated antibody against a mapped region if unexplained compartment differences persist (UniProt Q96Q04 isoforms; standard IHC validation practice).
How can I assess LMTK3 by IF alongside a neuronal marker?
Treat IF/ICC as a separately optimized application; the selected antibody evidence documents paraffin-section chromogenic IHC (datasheet A08432-2). Multiplex with an independently validated neuronal marker, then check whether LMTK3 signal follows neuronal cell bodies or projections as expected from tissue IHC (HPA: cytoplasmic and axonal neuronal expression; standard IF practice). Choose a far-red fluorophore, approximately 650 nm emission, and inspect an unstained section because tissue autofluorescence can confound shorter wavelengths (standard IF practice). Optimize permeabilisation only after the epitope and its membrane-facing side are established; residues 40–60 define a transmembrane segment but do not locate this antibody’s epitope (UniProt Q96Q04 topology; standard IF practice).
What should I check when LMTK3 DAB staining appears widespread?
Separate widespread chromogen from specific cellular signal by reviewing a no-primary control beside the test section (standard IHC practice). The documented workflow used 10% goat serum blocking, biotinylated secondary antibody, a streptavidin–biotin detection complex and DAB (datasheet A08432-2). Check peroxidase quenching and the no-primary control for endogenous enzyme signal; also assess endogenous biotin when using a biotin-based detection system (standard IHC practice; datasheet A08432-2). Reduce nonspecific staining by titrating antibody concentration and washing while preserving the documented citrate pH 6 retrieval condition; judge changes against neuronal cytoplasmic and axonal staining, not total brown area (datasheet A08432-2; HPA: tissue IHC; standard IHC practice).
How should I quantify LMTK3 staining across brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the neuronal population and anatomical region before scoring, since HPA reports high staining in caudate neuronal cells, cerebral cortex neuronal cells and cerebellar Purkinje cells (HPA: tissue IHC). For cell-body signal, record the percentage of positive neurons and an H-score on a 0–300 scale using fixed intensity categories (standard IHC scoring practice). For projection staining, measure positive axonal area or length per mm² of analyzed tissue with the same threshold across sections (HPA: axonal expression; standard image-analysis practice). Normalize cell-body results to counted neurons and projection results to viable analyzed area, excluding folds and damaged edges consistently (standard IHC practice).
When is a strong LMTK3 IHC signal likely to be artefactual?
A convincing brain pattern includes neuronal cytoplasmic or axonal staining, consistent with HPA tissue IHC and LMTK3’s annotated cell-projection localization (HPA: tissue IHC; UniProt Q96Q04 localisation). Question uniform staining in unrelated compartments or cells when it lacks this structure, while recognizing that LMTK3 staining also occurs outside brain (HPA: tissue IHC). Compare an adjacent no-primary section to identify detection background, particularly with the documented biotin-based DAB workflow (datasheet A08432-2; standard IHC practice). Exclude section edges, folds and necrotic regions from interpretation, and investigate diffuse brown signal that tracks endogenous enzyme activity rather than cell boundaries (standard IHC practice).
Boster reagents
Best LMTK3 / Serine/threonine-protein kinase LMTK3 IHC Antibodies
The catalog antibody has IHC data from paraffin sections of mouse and rat brain (A08432-2 image captions). Human reactivity is listed, but no IF/ICC data are supplied (A08432-2 catalog).
Real IHC data
Anti-LMTK3 Antibody ®
Cat # A08432-2
A08432-2 is the only SKU shown and lists IHC among its applications, with Human, Mouse and Rat reactivity (A08432-2 catalog). Its IHC captions show staining in paraffin sections of mouse and rat brain; the fixative is unreported (A08432-2 image captions).
Which to pick: Choose A08432-2 for paraffin section IHC: its own captions document mouse and rat brain staining, and its listed IHC dilution is 0.5–1 μg/ml (A08432-2 image captions; datasheet: 0.5–1 μg/ml). For cross-species work, it lists Human, Mouse and Rat reactivity, though the supplied IHC images document only mouse and rat tissue (A08432-2 catalog; A08432-2 image captions). No IF/ICC-validated SKU or IF/ICC image is supplied, so an IF/ICC choice cannot be supported here (A08432-2 applications and image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96Q04 (LMTK3_HUMAN, Serine/threonine-protein kinase LMTK3).
- Human Protein Atlas. LMTK3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LMTK3 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cell junctions..
- Human Protein Atlas. LMTK3 antibody validation summary (2 antibodies).
- Prognostic role of lemur tyrosine kinase-3 germline polymorphisms in adjuvant gastric cancer in Japan and the United States. Molecular cancer therapeutics 2013 — PMC3810398.
- LMTK3 confers chemo-resistance in breast cancer. Oncogene 2018 — PMC5992129.
- LMTK3 knockdown retards cell growth and invasion and promotes apoptosis in thyroid cancer. Molecular medicine reports 2017 — PMC5364963.
- Integrated multi-omics approach to distinct molecular characterization and classification of early-onset colorectal cancer. Cell reports. Medicine 2023 — PMC10040411.
- PubMed PMID:11572484 — UniProt-cited evidence.
- PubMed PMID:15057824 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LMTK3 (Serine/threonine-protein kinase LMTK3). Expected localisation Neuronal cytoplasm and axons (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A08432-2). Validated IHC antibody: A08432-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissues; fixative not specified (datasheet A08432-2); 2. Fixation: Image fixative and duration unreported (datasheet A08432-2); 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: Citrate pH 6 , 20 min (datasheet A08432-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08432-2); 8. Primary antibody: Rabbit anti-LMTK3, 0.5-1μg/ml (datasheet A08432-2); 9. Primary incubation: Overnight at 4 °C (datasheet A08432-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08432-2); Caveat: Scoring cell bodies alone can miss axonal staining (HPA tissue IHC); Isoform / epitope: No isoforms annotated; signal peptide 1–20 is cleaved, so check epitope position (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q96Q04, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08432-2, PMC3810398, PMC5364963. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LMTK3/immunohistochemistry-ihc-guide.