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LAMP3 / Lysosome-associated membrane glycoprotein 3 · IHC design guide
Design Immunohistochemistry for LAMP3
Plan LAMP3 IHC in paraffin sections around cytoplasmic staining in type II pneumocytes and non-germinal center lymphoid cells (HPA tissue IHC). This guide covers fixation, antibody titration, chromogenic detection, controls, and scoring by cell type and compartment (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LAMP3 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 lung and lymphoid cells (HPA tissue IHC) | |
| Staining pattern | Type II pneumocytes: high cytoplasmic; lymphoid cells: medium (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Lung+2 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 A09406) | |
| Caveat | Lymphoid staining may be weaker than type II pneumocyte staining (HPA tissue IHC) | |
| Regulation | Up-regulated in several carcinomas (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; lumenal versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended LAMP3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published LAMP3 staining protocols for oral squamous cell carcinoma specimens and pediatric gastric mucosa (PMC11491047; PMC6885256).
| Sample | Paraffin-embedded human lung-cancer tissue; fixative not specified (datasheet A09406) |
| Fixation | Image fixative and duration unreported (datasheet A09406); 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-LAMP3, 1:500 recommended; image 1:200 (datasheet A09406) |
| 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 | LAMP3-positive staining in alveolar cells type II of lung (HPA tissue IHC: High). HPA tissue profile: Selective cytoplasmic expression in type II pneumocytes and lymphoid non-germinal center cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval); the gastric mucosa protocol used high-pH retrieval (PMC6885256).
Section 2
What Is the Expected LAMP3 Staining Pattern?
LAMP3 is a membrane protein found on lysosomes, endosomes, cytoplasmic vesicles and the cell surface (UniProt Q9UQV4 topology and subcellular location). In IHC, expect cytoplasmic staining strongest in lung type II pneumocytes, with moderate staining in lymph node and tonsil non-germinal center cells (HPA: High; Medium). HPA rates its tissue pattern Enhanced, citing high agreement between antibody staining and RNA expression (HPA: Enhanced reliability).
| Distinct cytoplasmic staining in alveolar type II cells. | This matches the strongest reported normal-tissue signal (HPA: Lung, alveolar type II cells, High). A granular or vesicular appearance fits LAMP3 on intracellular membranes (UniProt Q9UQV4 subcellular location); HPA reports selective cytoplasmic IHC staining, without defining one required chromogenic texture (HPA: tissue IHC profile). |
| Staining confined mainly to nuclei, with little cytoplasmic signal. | Treat a nuclear-only pattern as suspect: the supported locations are vesicle, endosome and lysosome membranes, plus the cell surface (UniProt Q9UQV4 subcellular location). Check morphology and the known-positive control before scoring it as LAMP3; localization alone cannot identify the cause of an aberrant signal (general IHC practice). |
| Strong staining in cells outside the expected positive population. | In normal lung, identify type II pneumocytes before assigning the signal to LAMP3 (HPA: Lung, alveolar type II cells, High). Unexpected staining can reflect cross-reactivity or endogenous detection activity (general IHC practice). Interpret tumor tissue separately: UniProt reports LAMP3 up-regulation in several carcinomas (UniProt Q9UQV4 tissue specificity). |
| Weak, diffuse color across many structures and empty areas. | A broad haze does not reproduce the selective cytoplasmic tissue pattern (HPA: tissue IHC profile). Consider nonspecific antibody binding, inadequate blocking, excess detection reagent or incomplete washing (general IHC practice). Judge any localized signal against a matched control section after the background is addressed. |
| No detectable signal in a lung section expected to contain type II cells. | First verify that the section contains identifiable type II pneumocytes; these cells have High LAMP3 staining in HPA tissue IHC (HPA: Lung, alveolar type II cells, High). If present, review the validated IHC conditions and detection controls before concluding that the specimen lacks LAMP3 (general IHC practice). |
💡Expected LAMP3 appearanceCall a normal-tissue IHC result positive when type II pneumocytes show clear cytoplasmic staining, typically stronger than the Medium non-germinal center signal reported in lymph node or tonsil; diffuse haze or nuclear-only color is suspect (HPA: High in lung, Medium in lymph node and tonsil; UniProt Q9UQV4 subcellular location).
| Membrane location and topology | LAMP3 has one transmembrane segment at residues 382–402, a lumenal region at 28–381 and a short cytoplasmic tail at 403–416 (UniProt Q9UQV4 topology). Its intracellular membrane locations support cytoplasmic organelle-associated IHC staining; topology alone does not establish where a particular antibody binds. |
| Protein processing and glycosylation | The signal peptide spans residues 1–27, and the annotated mature chain starts at residue 28; seven glycosylation sites are listed (UniProt Q9UQV4 processing and glycosylation). These annotations describe the antigen, but provide no evidence here for epitope accessibility or an antigen-retrieval choice. |
| Tissue and cell-type distribution | Use cell identity when comparing sections: HPA reports High staining in lung type II cells and Medium staining in lymph node and tonsil non-germinal center cells (HPA: tissue IHC). Its Not detected calls apply to the specified cells sampled, such as adipocytes in adipose tissue, rather than every cell in that organ. |
| Strength of the IHC evidence | HPA calls the tissue profile Enhanced and lists two antibodies with Enhanced IHC validation, HPA051467 and CAB025133 (HPA: tissue IHC reliability; HPA: antibody validation). This supports the reported tissue pattern, but does not validate every staining condition or establish target-specific fixation sensitivity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Lung control is blank. | The section may lack identifiable type II cells, or an IHC step may have failed (HPA: High in lung type II cells; general IHC practice). | Confirm cell morphology, then review the catalog antibody’s IHC-P conditions, reagent activity and detection controls (general IHC practice). Do not use a blank, cell-poor field to call the target absent. |
| Lymphoid staining seems weaker than lung. | That difference matches the reported levels: Medium in lymph node and tonsil non-germinal center cells versus High in lung type II cells (HPA: tissue IHC). | Compare the correct cell populations and score each tissue against its expected level (HPA: tissue IHC; general IHC practice). Avoid requiring lung-level intensity in lymphoid sections. |
| Color appears mainly in nuclei. | Nuclear localization is outside the listed LAMP3 compartments (UniProt Q9UQV4 subcellular location). | Review the counterstain and chromogen localization, and compare with a known-positive control and an appropriate detection control (general IHC practice). Reassess the signal before scoring it as specific. |
| Many unrelated cells or blank areas are colored. | Nonspecific binding, endogenous detection activity or excess reagent can create misleading color (general IHC practice). | Check blocking, washing and detection controls; review antibody concentration within its validated IHC-P guidance (general IHC practice). Look for the selective cytoplasmic pattern after background is reduced (HPA: tissue IHC profile). |
| A listed negative cell type stains. | HPA reports Not detected in particular cells, including adipocytes in adipose tissue; that designation is cell-specific (HPA: tissue IHC). | Confirm the stained cell’s identity and compare a detection control (general IHC practice). Investigate persistent signal as possible cross-reactivity or endogenous activity before treating the HPA pattern as contradicted. |
| IF/ICC: what signal should appear? | HPA reports vesicles as the enhanced ICC-IF location, consistent with LAMP3 on cytoplasmic vesicle membranes (HPA: subcellular location; UniProt Q9UQV4 subcellular location). | Expect intracellular vesicular signal and assess it with appropriate IF controls (HPA: subcellular location; general IF practice). Use the separate IF/ICC guide for assay setup; this IHC-P section supplies no IF protocol. |
Experiment evidence
IHC & IF images showing LAMP3 staining
A09406 · IHCImmunohistochemical analysis of paraffin-embedded human-lung-cancer, antibody was diluted at 1:200
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000078081-LAMP3/tissue/cerebral+cortex#imid_6560427
HPA · LungHuman Protein Atlas IHC, Lung. Source: https://www.proteinatlas.org/ENSG00000078081-LAMP3/tissue/lung#imid_6555847
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000078081-LAMP3/tissue/colon#imid_6555894
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000078081-LAMP3/tissue/liver#imid_15292993
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB025133; green = LAMP3, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB025133; green = LAMP3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LAMP3 IHC & IF
🧪Run lung first: alveolar type II cells should stain (HPA: High in alveolar cells type II). Use adipose tissue as the negative tissue, assessing adipocytes (HPA: Not detected in adipocytes); on the lung slide, morphologically distinct cells without specific staining can serve as internal negative references, but their absence of LAMP3 is not established by the supplied HPA row.
Positive control tissue: Lung (Alveolar cells type II, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LAMP3 in U-251MG, A-431, HEK293, U2OS, with annotated localisation: Vesicles (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; use an irrelevant antibody matched to the primary’s host and isotype for a monoclonal, or host-matched nonimmune IgG for a polyclonal, and validate specificity with LAMP3 knockout material or peptide competition when the immunizing peptide is available (standard IHC practice). For lung sections, quench endogenous peroxidase for chromogenic detection and assess tissue pigment or autofluorescence when interpreting IHC or IF signal (standard IHC/IF practice).
⚠️Feasibility: No supplied source reports a LAMP3-specific fixation window or fixation effect, and the fixative in the selected paraffin-section caption is unreported (SKU A09406 caption). Retrieval requirements for LAMP3 are unreported; optimize antigen retrieval empirically for paraffin IHC (standard IHC practice). Vesicular ICC-IF staining is documented (HPA: subcellular localization), but the evidence does not establish whether frozen sections or IF are easier than paraffin IHC; lung pigment and autofluorescence can complicate interpretation (standard IHC/IF practice).
HPA tissue IHC evidence for LAMP3
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 |
|---|---|---|---|---|
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LAMP3 IHC Tips
Troubleshoot LAMP3 staining in paraffin sections using the catalog antibody’s tissue image, the stated retrieval setting, and expected cellular distribution.
What should I change when LAMP3 staining is weak after antigen retrieval?
For paraffin section IHC, start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Check that sections remain attached and compare retrieval runs using the same antibody dilution and detection conditions (standard IHC practice). The catalog image shows staining of paraffin-embedded human lung cancer at 1:200, but its caption does not report a fixative or retrieval method (A09406 caption). If staining remains weak, adjust heating duration in a controlled comparison and include a reference section with the expected cellular pattern (standard IHC practice; HPA: selective cytoplasmic expression in type II pneumocytes and lymphoid non-germinal center cells).
Could fixation explain inconsistent LAMP3 staining between paraffin blocks?
Target-specific LAMP3 sensitivity to fixation is unknown from the supplied evidence; do not infer it from tissue staining patterns or protein topology (A09406 caption; HPA tissue IHC; UniProt Q9UQV4 topology). The catalog image documents a paraffin-embedded human lung cancer section stained at 1:200, but does not state its fixative (A09406 caption). Record fixative, fixation duration, block age and processing history for each specimen, then compare sections using citrate pH 6.0 retrieval under identical detection conditions (standard IHC practice; page retrieval rule). Include comparable reference tissue in each run, and interpret differences between blocks cautiously until processing variables have been controlled (standard IHC practice).
Where should convincing LAMP3 signal appear in a tissue section?
Expect chiefly cytoplasmic, often punctate staining in appropriate cells, consistent with the tissue and vesicle observations (HPA tissue IHC: selective cytoplasmic expression; HPA subcellular: vesicles). LAMP3 is reported at lysosome, early endosome and cytoplasmic vesicle membranes, and also at the cell surface (UniProt Q9UQV4 localisation). In lung, assess alveolar type II cells; in lymph node and tonsil, assess non-germinal center cells (HPA tissue IHC). Compare staining in these cells with surrounding cells on the same section before changing the 1:200 dilution used in the catalog image (HPA tissue IHC; A09406 caption; standard IHC practice).
How could LAMP3 topology affect an antibody’s apparent staining pattern?
LAMP3 has a lumenal region at residues 28–381, one transmembrane segment at 382–402, and a cytoplasmic tail at 403–416 (UniProt Q9UQV4 topology). Seven glycosylation sites are annotated in its lumenal region, so epitope accessibility may depend on the antibody’s binding site and processing conditions (UniProt Q9UQV4 glycosylation; standard IHC practice). No isoforms are annotated in the supplied record; this does not identify the catalog antibody’s epitope (UniProt Q9UQV4 isoforms; A09406 caption). If staining differs across blocks, compare matched sections with citrate pH 6.0 retrieval before attributing the difference to an isoform or glycosylation (page retrieval rule; standard IHC practice).
How should I check an IHC LAMP3 pattern by multiplex immunofluorescence?
Use a validated marker for the expected cell type alongside LAMP3, such as a type II pneumocyte marker when examining lung, and assess cellular overlap (HPA tissue IHC; standard IF practice). Choose a spectrally separated, preferably far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and inspect single-label controls (standard IF practice). If the antibody binds a lumenal epitope, permeabilisation must allow access to intracellular vesicles; the supplied catalog caption does not identify its epitope or establish an IF condition (UniProt Q9UQV4 topology; A09406 caption; standard IF practice). Treat the paraffin IHC dilution of 1:200 as image context, then optimise IF staining and controls separately (A09406 caption; standard IF practice).
How can I reduce diffuse brown staining without losing LAMP3 signal?
Compare a no-primary control with the LAMP3 section to identify detection background, and inspect tissue edges and damaged areas separately (standard IHC practice). Apply an endogenous peroxidase block before chromogenic detection and use an appropriate blocking step for the detection system (standard IHC practice). Keep citrate pH 6.0 retrieval and the catalog image’s 1:200 dilution documented while changing one staining variable at a time (page retrieval rule; A09406 caption; standard IHC practice). Preserve staining in the expected cell populations as the practical reference when reducing background (HPA tissue IHC: type II pneumocytes and lymphoid non-germinal center cells; standard IHC practice).
What should I score when comparing LAMP3 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; type II pneumocytes and lymphoid non-germinal center cells have documented LAMP3 staining (HPA tissue IHC; standard IHC practice). Within that region, report percentage of positive cells and staining intensity, or calculate an H-score using intensity grades 0–3 (standard IHC practice). For sparse positive cells, report their density per mm² of viable tissue and state how regions were selected (standard IHC practice). Normalise counts to the relevant cell population or viable tissue area, and keep retrieval, detection and image thresholds consistent across sections (standard IHC practice; page retrieval rule).
How do I distinguish genuine LAMP3 staining from tissue artefact?
A convincing result aligns with cytoplasmic or vesicular staining in an expected cell population (HPA tissue IHC; HPA subcellular: vesicles). In lung, evaluate type II pneumocytes; in lymphoid tissue, evaluate non-germinal center cells rather than assigning every stained cell the same identity (HPA tissue IHC; standard IHC practice). Treat isolated nuclear signal, staining confined to section edges or necrotic areas, and color in a no-primary control as reasons to investigate artefact (UniProt Q9UQV4 localisation; standard IHC practice). Check endogenous peroxidase background and compare a reference section processed with citrate pH 6.0 before interpreting a changed pattern biologically (standard IHC practice; page retrieval rule).
Boster reagents
Best LAMP3 / Lysosome-associated membrane glycoprotein 3 IHC Antibodies
The rendered LAMP3 antibody has an IHC image from paraffin-embedded human lung cancer (A09406 IHC image caption); IHC and IF are listed for human, mouse and rat (A09406 catalog applications and reactivity).
Real IHC data
Anti-DC-LAMP LAMP3 Antibody
Cat # A09406
A09406 is the only SKU with a rendered card, and its IHC image shows paraffin-embedded human lung cancer (A09406 IHC image caption). Its catalog lists IHC and IF for human, mouse and rat, but provides no IF image (A09406 catalog applications, reactivity and image alts).
Which to pick: For tissue IHC, pick A09406 when its documented paraffin-section example is useful; the image caption does not report a fixative (A09406 IHC image caption). For IF/ICC, A09406 lists IF, while ICC validation and an IF image are unreported (A09406 catalog applications and image alts). For monkey tissue IHC, consider A09406-3: it lists monkey reactivity and IHC and is polyclonal, but has no IHC image; both SKUs list human, mouse and rat reactivity (A09406-3 catalog reactivity, applications, dilution_raw and image alts; A09406 catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UQV4 (LAMP3_HUMAN, Lysosome-associated membrane glycoprotein 3).
- Human Protein Atlas. LAMP3 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LAMP3 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. LAMP3 antibody validation summary (2 antibodies).
- Lysosome-Associated Membrane Protein-3 (LAMP3) Expression in Oral Squamous Cell Carcinoma and Its Relationship With Clinicopathological Parameters: A Cross-Sectional Study. Cureus 2024 — PMC11491047.
- Relationship between Helicobacter pylori Infection and Plasmacytoid and Myeloid Dendritic Cells in Peripheral Blood and Gastric Mucosa of Children. Mediators of inflammation 2019 — PMC6885256.
- Recessive missense LAMP3 variant associated with defect in lamellar body biogenesis and fatal neonatal interstitial lung disease in dogs. PLoS genetics 2020 — PMC7082050.
- Development and Validation of a 7-Gene Inflammatory Signature Forecasts Prognosis and Diverse Immune Landscape in Lung Adenocarcinoma. Frontiers in molecular biosciences 2022 — PMC8964604.
- PubMed PMID:9721848 — UniProt-cited evidence.
- PubMed PMID:9768752 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LAMP3 (Lysosome-associated membrane glycoprotein 3). Expected localisation Cytoplasmic staining in lung and lymphoid cells (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A09406 at 1:500 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Alveolar cells type II); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung-cancer tissue; fixative not specified (datasheet A09406); 2. Fixation: Image fixative and duration unreported (datasheet A09406); 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-LAMP3, 1:500 recommended; image 1:200 (datasheet A09406); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A09406); Caveat: Lymphoid staining may be weaker than type II pneumocyte staining (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; lumenal versus cytoplasmic epitope matters (UniProt); Regulation: Up-regulated in several carcinomas (UniProt). Evidence: UniProt Q9UQV4, HPA tissue IHC · Enhanced, HPA ICC-IF · vesicles., Datasheet A09406, PMC11491047, PMC6885256. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LAMP3/immunohistochemistry-ihc-guide.