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LAMTOR3 / Ragulator complex protein LAMTOR3 · IHC design guide
Design Immunohistochemistry for LAMTOR3
Plan paraffin-section IHC for LAMTOR3 using its granular cytoplasmic tissue pattern (HPA tissue IHC). Compare high-staining lung macrophages with adipocytes, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LAMTOR3 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 | Granular cytoplasm (HPA tissue IHC); late endosome membrane (UniProt) | |
| Staining pattern | Granular cytoplasm in lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+3 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 | Adipocytes show no detected staining (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | Two isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended LAMTOR3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published LAMTOR3 IHC protocol using human kidney specimens (PMC9459259).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A08040); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-LAMTOR3, 5 μg/mL (datasheet A08040) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LAMTOR3-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min, consistent with this page’s retrieval rule for membrane-associated antigens (UniProt Q9UHA4 localization).
Section 2
What Is the Expected LAMTOR3 Staining Pattern?
LAMTOR3 is recruited by LAMTOR1 to late endosome and lysosome membranes and has no transmembrane segment (UniProt Q9UHA4). In paraffin sections, expect granular cytoplasmic staining in appropriate cells, including lung macrophages, adrenal glandular cells and cerebral cortical neurons reported as high by HPA (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Granular cytoplasmic staining in lung macrophages or adrenal glandular cells (HPA tissue IHC). | This matches HPA's general cytoplasmic, granular profile and its High ratings for those cells (HPA tissue IHC). The granules are consistent with endolysosomal localisation, but a chromogenic pattern alone cannot identify individual organelles (UniProt Q9UHA4; general IHC practice). |
| Predominantly nuclear staining, with little granular cytoplasmic signal. | Treat this as a compartment mismatch: UniProt places LAMTOR3 at late endosome membranes, and HPA describes granular cytoplasm (UniProt Q9UHA4; HPA tissue IHC). Review the counterstain, detection controls and staining conditions before scoring nuclei as positive (general IHC practice). |
| Strong staining chiefly in adipocytes or skeletal myocytes. | HPA reports LAMTOR3 as Not detected in those cells, so this is discordant with its tissue observations (HPA tissue IHC). Cross-reactivity or endogenous detection activity is possible; morphology alone cannot distinguish them (general IHC practice). |
| Broad, diffuse colour across cells and surrounding tissue, obscuring granules. | The expected HPA profile is granular cytoplasm, so diffuse background makes cell-level interpretation unreliable (HPA tissue IHC). Check reagent-only controls and whether blocking, washing or chromogen development contributed to nonspecific colour (general IHC practice). |
| No staining in lung macrophages or another HPA High cell population. | A blank expected-positive population warrants checking the run before calling LAMTOR3 absent (HPA tissue IHC; general IHC practice). HPA's High rating describes observed tissue staining, not a guarantee that every section or assay will be positive (HPA tissue IHC). |
💡Expected LAMTOR3 appearanceCall a positive result when relevant cells show clear granular cytoplasmic staining, especially HPA High populations such as lung macrophages; predominantly nuclear colour or strong staining in HPA Not detected cell types is suspect (HPA tissue IHC; UniProt Q9UHA4).
| Subcellular placement | LAMTOR1 recruits LAMTOR3 to late endosome and lysosome membranes, although LAMTOR3 has no transmembrane segment (UniProt Q9UHA4). Score the observed cytoplasmic pattern; do not require a crisp cell-surface membrane outline (HPA tissue IHC; UniProt Q9UHA4). |
| Tissue and cell choice | HPA reports High staining in several distinct populations, including lung macrophages, testicular Leydig cells and prostate glandular cells, while adipocytes are Not detected (HPA tissue IHC). Use cell identity when comparing fields; the record does not support calling an entire tissue uniformly positive or negative (HPA tissue IHC). |
| Strength of IHC evidence | The tissue profile is Supported with medium staining-to-RNA consistency; antibody HPA026858 is also IHC Supported, with no Enhanced designation supplied (HPA tissue IHC; HPA antibodies). Interpret an unexpected pattern as a finding to verify, rather than proof of a new localisation (HPA tissue IHC; general IHC practice). |
| Isoforms and processing | UniProt lists 2 isoforms and a chain spanning residues 1–124, with no signal peptide or propeptide annotated (UniProt Q9UHA4). The payload supplies no antibody epitope, so it cannot establish whether the IHC reagent detects both isoforms (UniProt Q9UHA4; HPA antibodies). |
| IF/ICC: what should be expected? | For IF/ICC, UniProt supports an endolysosomal association, but HPA provides no main ICC-IF location or cell-line images here (UniProt Q9UHA4; HPA subcellular). Therefore this IHC pattern cannot be presented as an observed IF/ICC pattern (HPA tissue IHC; HPA subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| An HPA High cell population is blank (HPA tissue IHC). | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Confirm the expected cell type is present, then review a positive control, reagent sequence, retrieval and detection performance under the assay's general IHC workflow (general IHC practice). |
| Signal is mainly nuclear instead of granular cytoplasmic (HPA tissue IHC). | The compartment conflicts with the reported localisation; counterstain confusion or nonspecific signal is possible (UniProt Q9UHA4; HPA tissue IHC; general IHC practice). | Compare with a detection control and inspect matched cell morphology before assigning nuclear positivity (general IHC practice). |
| Strong colour appears in adipocytes or skeletal myocytes (HPA tissue IHC). | Those cells are Not detected by HPA; cross-reactivity or endogenous detection activity may explain discordant colour (HPA tissue IHC; general IHC practice). | Check a reagent-only or primary-omission control, then assess whether the signal remains specific to the expected cells (general IHC practice; HPA tissue IHC). |
| Diffuse background hides the cytoplasmic granules (HPA tissue IHC). | Nonspecific reagent binding, insufficient washing or excessive chromogen development can obscure cell-level staining (general IHC practice). | Use the run's background controls and review blocking, wash steps and development time; score only when the granular cell pattern can be distinguished (general IHC practice; HPA tissue IHC). |
| A field looks uniformly positive despite mixed cell types. | HPA reports cell-specific High and Not detected observations, so uniform colour may reflect background or an overbroad reading of the section (HPA tissue IHC; general IHC practice). | Identify cells by morphology, compare expected and Not detected populations, and record staining by cell type rather than by whole-section colour (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing LAMTOR3 staining
A08040 · IHCImmunohistochemistry of LAMTOR3 in human brain tissue with LAMTOR3 antibody at 5 μg/mL.
A08040 · IFImmunofluorescence of LAMTOR3 in human brain tissue with LAMTOR3 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109270-LAMTOR3/tissue/cerebral+cortex#imid_6874464
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109270-LAMTOR3/tissue/colon#imid_6874398
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109270-LAMTOR3/tissue/liver#imid_6874399
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000109270-LAMTOR3/tissue/kidney#imid_6874400
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LAMTOR3 IHC & IF
🧪Run lung first: macrophages should stain (HPA: High in lung macrophages). Use adipose tissue as the negative tissue: adipocytes should remain at background (HPA: Not detected in adipocytes); on the lung slide, assess non-macrophage cells against the no-primary control rather than assuming they are LAMTOR3-negative.
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 carries no ICC-IF cell line for LAMTOR3; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and concentration-matched isotype controls matched to the primary antibody’s host species and isotype, plus LAMTOR3 knockout material or a peptide-block control if the immunizing peptide is available (standard IHC practice). In lung, check endogenous peroxidase and macrophage pigment against the no-primary slide before scoring chromogenic signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A08040 brain IHC caption does not state a fixative (caption: fixative not stated). Retrieval dependency is unreported; compare retrieval conditions on paraffin sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; lung macrophage pigment and endogenous peroxidase can complicate chromogenic scoring (standard IHC practice).
HPA tissue IHC evidence for LAMTOR3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 | Glial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Lung | Macrophages | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LAMTOR3 IHC Tips
Troubleshoot LAMTOR3 staining by checking retrieval, controls, and cellular pattern before interpreting chromogenic signal (UniProt Q9UHA4; HPA tissue IHC).
Which retrieval conditions should I try first for LAMTOR3 paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 minutes (page retrieval setting). Cool sections in the buffer, then compare the same antibody dilution and detection conditions across runs so retrieval is the main variable (standard IHC practice). If staining remains weak, test a different retrieval buffer on matched sections as a fallback, while checking whether tissue structure and background deteriorate (standard IHC practice). Score the expected granular cytoplasmic signal rather than diffuse stain alone (HPA tissue IHC: general cytoplasmic expression with a granular pattern). Include a no-primary control to expose detection background (standard IHC practice).
Could fixation explain weak or uneven LAMTOR3 staining?
LAMTOR3-specific sensitivity to fixation is unknown from the supplied evidence (selected A08040 tissue-IHC caption: fixative not stated). Record the fixative, time before fixation, fixation duration, and section age for each specimen before comparing staining intensity (standard IHC practice). On matched sections, keep citrate pH 6.0 retrieval at 95–98 °C for 20 minutes constant while comparing specimens with different documented processing histories (page retrieval setting; standard IHC practice). Check morphology alongside signal: damaged or detached areas cannot support reliable cellular scoring (standard IHC practice). Do not assign a LAMTOR3-specific fixation effect from its reported localisation or tissue staining pattern (UniProt Q9UHA4; HPA tissue IHC).
What LAMTOR3 staining pattern is plausible in tissue sections?
Expect predominantly granular cytoplasmic staining in chromogenic sections (HPA tissue IHC: general cytoplasmic expression with a granular pattern). LAMTOR3 is recruited by LAMTOR1 to late endosome and lysosome membranes and has no transmembrane segment (UniProt Q9UHA4 localisation and topology). At light-microscope resolution, assess cytoplasmic granularity and cellular context; do not identify individual puncta as lysosomes from DAB staining alone (UniProt Q9UHA4 localisation; standard IHC interpretation). Compare glandular cells in adrenal gland or lung macrophages as reported high-staining contexts, with matched controls (HPA: High in adrenal gland glandular cells; High in lung macrophages). Predominantly nuclear staining warrants specificity checks before interpretation (UniProt Q9UHA4 localisation; standard IHC practice).
How should I assess an antibody when its LAMTOR3 epitope is unspecified?
LAMTOR3 has 2 listed isoforms, and the supplied record does not map the catalog antibody’s epitope to either one (UniProt Q9UHA4 isoforms; supplied antibody evidence). Do not claim isoform-specific IHC staining without an epitope map and supporting validation (standard IHC interpretation). Its annotated chain spans residues 1–124, with no annotated glycosylation sites or modified residues; these annotations do not establish which epitope survives tissue processing (UniProt Q9UHA4 processing and PTMs). Compare staining across matched positive and low-staining cell populations, then seek independent specificity evidence if interpretation depends on a particular isoform (HPA tissue IHC; standard IHC practice). Keep retrieval and detection settings identical during that comparison (standard IHC practice).
How can IF help check the LAMTOR3 pattern seen by chromogenic IHC?
Treat IF as a separate assay requiring its own antibody and processing validation; the supplied HPA subcellular record provides no ICC/IF images (HPA subcellular). Multiplex LAMTOR3 with a marker identifying the expected cell type, such as macrophages in lung, and assess whether signal falls within those cells (HPA: High in lung macrophages; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and include unstained and single-label controls before judging puncta (standard IF practice). Because LAMTOR3 is recruited to endosome and lysosome membranes without a transmembrane segment, optimise gentle permeabilisation for access to its antibody epitope, whose orientation is unspecified here (UniProt Q9UHA4 topology and localisation; standard IF practice).
What should I adjust when LAMTOR3 DAB staining obscures cellular detail?
First compare a no-primary section with the stained section to distinguish detection-system signal from antibody-dependent staining (standard IHC practice). Quench endogenous peroxidase before DAB detection, apply an appropriate protein block, and titrate the primary antibody and DAB development time on matched sections (standard chromogenic IHC practice). The A08040 brain-tissue caption reports 5 µg/mL antibody but does not state a fixative or establish that concentration for every tissue (selected A08040 tissue-IHC caption). Judge optimisation against the reported granular cytoplasmic pattern, while watching for diffuse deposits and section-edge signal (HPA tissue IHC; standard IHC practice). Record exposure and processing conditions with each comparison (standard IHC practice).
How should I quantify LAMTOR3 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, using morphology or a validated cell-type marker where needed (standard IHC practice). Report an H-score from staining intensity and percentage of positive cells, or report the percentage of positive cells within the annotated population (standard IHC quantification). For granular signal, a positive-cell density per mm² can supplement those measures if the counted tissue area is recorded (HPA tissue IHC: granular cytoplasmic pattern; standard IHC quantification). Normalise counts to evaluable cells or viable tissue area, and compare sections processed with the same retrieval, detection, and scoring thresholds (standard IHC practice). Exclude folds, edges, and necrotic areas using predefined rules (standard IHC practice).
When should I doubt an apparent LAMTOR3-positive IHC result?
A convincing result shows cellular, granular cytoplasmic signal in an appropriate tissue context and exceeds the no-primary control (HPA tissue IHC; standard IHC practice). High staining is reported in lung macrophages and cerebral-cortex neuronal cells, whereas adipocytes are reported as not detected; use these as context, not absolute controls for every specimen (HPA tissue IHC). Question predominantly nuclear signal, isolated section-edge deposits, or stain confined to necrotic tissue (UniProt Q9UHA4 localisation; standard IHC interpretation). Persistent DAB signal without primary antibody suggests endogenous enzyme or detection background that needs troubleshooting (standard chromogenic IHC practice). The supplied brain-tissue image caption states the antibody concentration, but not its fixative (selected A08040 tissue-IHC caption).
Boster reagents
Best LAMTOR3 / Ragulator complex protein LAMTOR3 IHC Antibodies
A08040 has IHC-P and IF applications with human brain tissue images; listed reactivity covers human, mouse and rat (catalog: applications, image captions and reactivity).
Real IHC data
Anti-LAMTOR3 Antibody
Cat # A08040
A08040 has an IHC image of human brain tissue at 5 μg/mL (A08040 IHC caption). A08040 also has an IF image of human brain tissue at 20 μg/mL; human, mouse and rat are listed as reactive species (A08040 IF caption; catalog: reactivity).
Which to pick: Choose A08040 for paraffin-section IHC because IHC-P is listed and its IHC caption shows staining in human brain tissue at 5 μg/mL; the fixative is unreported (catalog: applications; A08040 IHC caption). For IF, A08040 has a human brain tissue image at 20 μg/mL; ICC validation is unreported (A08040 IF caption; catalog: applications). For mouse or rat tissue, A08040 lists reactivity with both species, though its supplied IHC and IF images show human tissue; the antibody is rabbit-hosted, and clonality is unreported (catalog: reactivity, host and clone; A08040 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UHA4 (LTOR3_HUMAN, Ragulator complex protein LAMTOR3).
- Human Protein Atlas. LAMTOR3 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. LAMTOR3 subcellular location (ICC-IF): Highest expression in RPTEC/TERT1: 51.1 nTPM.
- Human Protein Atlas. LAMTOR3 antibody validation summary (1 antibodies).
- LAMTOR3 is a prognostic biomarker in kidney renal clear cell carcinoma. Journal of clinical laboratory analysis 2022 — PMC9459259.
- Circ_0075829 facilitates the progression of pancreatic carcinoma by sponging miR-1287-5p and activating LAMTOR3 signalling. Journal of cellular and molecular medicine 2020 — PMC7753824.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LAMTOR3 (Ragulator complex protein LAMTOR3). Expected localisation Granular cytoplasm (HPA tissue IHC); late endosome membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08040 at 5 μg/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: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A08040); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-LAMTOR3, 5 μg/mL (datasheet A08040); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Adipocytes show no detected staining (HPA tissue IHC); Isoform / epitope: Two isoforms; epitope coverage is unspecified (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q9UHA4, HPA tissue IHC · Supported, HPA cell-line RNA · Highest expression in RPTEC/TERT1: 51.1 nTPM, Datasheet A08040, PMC9459259. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LAMTOR3/immunohistochemistry-ihc-guide.