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LMAN1 / Protein ERGIC-53 · IHC design guide
Design Immunohistochemistry for LMAN1
Plan LMAN1 IHC in paraffin sections around the cytoplasmic staining reported across tissues (HPA tissue IHC). Compare high-staining glandular or endocrine cells with low-staining squamous epithelia, using matched fixation and cellular intensity scoring (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LMAN1 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 | Tissue cytoplasm (HPA tissue IHC); ER/ERGIC/Golgi membranes (UniProt) | |
| Staining pattern | Cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03628-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03628-1) | |
| Caveat | Low staining in some squamous epithelia despite broad expression (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | No isoforms listed; cleaved signal; luminal vs cytoplasmic epitopes matter (UniProt) |
Section 1
Recommended LMAN1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A03628-1) with a published LMAN1 IHC method using paraffin-embedded animal organ sections (PMC13011374).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03628-1) |
| Fixation | Image fixative and duration unreported (datasheet A03628-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03628-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03628-1) |
| Primary antibody | Rabbit anti-LMAN1, 2-5μg/ml (datasheet A03628-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03628-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03628-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LMAN1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A03628-1); the published method does not specify its antigen-unmasking conditions (PMC13011374).
Section 2
What Is the Expected LMAN1 Staining Pattern?
LMAN1 is an ER–Golgi trafficking protein in ER–Golgi intermediate compartment, Golgi, and ER membranes, with one transmembrane segment and a lumenal domain (UniProt P49257 localization and topology). On paraffin sections, expect cytoplasmic staining across tissues, with high staining reported in selected glandular and endocrine cells (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in glandular or endocrine cells, with stronger signal in an HPA high-staining tissue. | Consistent with LMAN1 IHC: HPA reports cytoplasmic expression across tissues and High staining in adrenal glandular and colon endocrine cells (HPA tissue IHC). Judge intensity against cells on the same slide; high staining in one cell group does not require equal intensity in every cell (HPA tissue IHC). |
| Signal appears predominantly nuclear, or outlines the cell surface without appreciable cytoplasmic staining. | This conflicts with the reported cytoplasmic tissue pattern and intracellular membrane locations (HPA tissue IHC; UniProt P49257 localization). Treat it as suspect and compare with an HPA high-staining tissue before assigning it to LMAN1 (general IHC practice). |
| A cell population stains intensely while expected glandular or endocrine cells remain unstained. | Consider antibody cross-reactivity or endogenous detection activity (general IHC practice). LMAN1 is broadly expressed, so staining outside one favored cell type alone is insufficient evidence of error (UniProt P49257 tissue specificity; HPA tissue IHC). Assess compartment and detection controls together. |
| Broad, diffuse chromogen obscures cell boundaries and the expected intracellular pattern. | A pattern that cannot be assigned to cells or cytoplasm is uninterpretable (general IHC practice). HPA reports cytoplasmic staining, but its Supported rating has only medium consistency with RNA expression; diffuse background should not be counted as confirmed LMAN1 signal (HPA tissue IHC). |
| Little or no signal appears in adrenal glandular or colon endocrine cells. | Those are reported High-staining examples, so absence raises concern about assay performance or specimen quality (HPA tissue IHC; general IHC practice). It does not, by itself, establish biological loss: HPA supplies no negative tissue reference here, and its tissue assessment is Supported with medium consistency (HPA tissue IHC). |
💡Expected LMAN1 appearanceCall a result positive when cellular cytoplasmic staining is evident, especially at high intensity in reported adrenal glandular or colon endocrine cells (HPA tissue IHC); predominantly nuclear, surface-only, or indiscriminate diffuse signal is suspect against the reported localization (HPA tissue IHC; UniProt P49257 localization).
| Intracellular location and topology | LMAN1 occupies ER–Golgi intermediate compartment, Golgi, and ER membranes; residues 31–477 are lumenal and 499–510 cytoplasmic (UniProt P49257 topology). An antibody's epitope is not supplied, so topology alone cannot predict its staining intensity or retrieval response. |
| Tissue and cell context | HPA describes cytoplasmic expression in all tissues and low tissue specificity, with High staining in selected glandular and endocrine cells and Low staining in listed populations such as smooth muscle cells (HPA tissue IHC). Low staining is a poor substitute for a documented negative control. |
| Antibody evidence | HPA lists IHC as Supported for HPA002320 and CAB037163; its overall tissue assessment is Supported with medium staining–RNA consistency (HPA antibodies; HPA tissue IHC). These ratings support pattern comparison without guaranteeing specificity for every specimen. |
| IF/ICC: what localization is reported? | Mainly vesicles, with additional ER localization, are Supported in HPA ICC-IF (HPA subcellular). This answers the localization question; paraffin chromogenic IHC should be interpreted against HPA's tissue cytoplasmic pattern (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High-staining cells show no chromogen. | A failed detection step or insufficient assay signal is possible (general IHC practice); the missing signal does not prove LMAN1 absence. | Check the IHC-validated antibody's documented dilution and detection conditions, then repeat with a reported High-staining tissue (general IHC practice; HPA tissue IHC). |
| Signal is mainly nuclear or confined to cell edges. | The distribution disagrees with cytoplasmic tissue staining and ER–Golgi or ER membrane localization (HPA tissue IHC; UniProt P49257 localization). | Review morphology and counterstain, and compare the compartment in a reported High-staining tissue before scoring positivity (general IHC practice; HPA tissue IHC). |
| Chromogen appears where the primary antibody was omitted. | Endogenous detection activity or reagent background may contribute signal (general IHC practice). | Address endogenous activity using controls and detection-system guidance; score only signal above the omission control (general IHC practice). |
| Most of the section has uniform haze. | Nonspecific background may hide cell-level cytoplasmic staining (general IHC practice; HPA tissue IHC). | Review blocking, antibody concentration, washes, and chromogen development; reassess cell boundaries and cytoplasm after background improves (general IHC practice). |
| Only a low-staining reference population is available. | HPA lists some Low populations but no negative tissue; a weak result there cannot establish assay failure or specificity (HPA tissue IHC). | Add a reported High-staining glandular or endocrine example for pattern comparison and retain an omission control for detection background (HPA tissue IHC; general IHC practice). |
| Staining varies between otherwise comparable paraffin sections. | Section handling or assay conditions may vary (general IHC practice). No LMAN1-specific fixation sensitivity is established by these sources. | Compare processing records and run sections together under the documented IHC conditions; avoid attributing variation specifically to LMAN1 epitope masking without evidence (general IHC practice). |
Experiment evidence
IHC & IF images showing LMAN1 staining
A03628-1 · IHCIHC analysis of LMAN1 using anti-LMAN1 antibody (A03768-1). LMAN1 was detected in a paraffin-embedded section of human lung cancer tissue. H
A03628-1 · IHCIHC analysis of LMAN1 using anti-LMAN1 antibody (A03768-1). LMAN1 was detected in a paraffin-embedded section of human spleen tissue. Heat m
A03628-1 · IFIF analysis of LMAN1 using anti-LMAN1 antibody (A03628-1). LMAN1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000074695-LMAN1/tissue/cerebral+cortex#imid_10406363
HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000074695-LMAN1/tissue/thyroid+gland#imid_10404723
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000074695-LMAN1/tissue/colon#imid_10342735
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000074695-LMAN1/tissue/liver#imid_10342728
HPA · ICC-IF EFO-21Human Protein Atlas immunofluorescence, EFO-21, antibody CAB037163; green = LMAN1, blue = nucleus, red = microtubules.
HPA · ICC-IF EFO-21Human Protein Atlas immunofluorescence, EFO-21, antibody CAB037163; green = LMAN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LMAN1 IHC & IF
🧪Run adrenal gland first and assess its glandular cells, which show High LMAN1 staining (HPA: adrenal gland glandular cells, High). HPA detects LMAN1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and expect any comparatively unstained cells on the positive slide to show only background signal rather than treating them as validated biological negatives (HPA: no negative tissues).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: LMAN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LMAN1 in EFO-21, U2OS, with annotated localisation: Vesicles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and a section treated with matched nonimmune rabbit IgG, alongside LMAN1 knockout material if available (caption: rabbit anti-LMAN1 antibody; standard IHC controls). For chromogenic staining, block endogenous peroxidase and inspect the adrenal section for native pigment that could resemble DAB signal (HPA: adrenal gland positive; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected paraffin-section caption does not state a fixative (selected tissue-IHC caption: fixative unreported). The caption uses heat retrieval in EDTA, pH 8.0, but does not establish whether retrieval is required for other specimens (selected tissue-IHC caption: EDTA retrieval). Frozen sections cannot be judged easier from this evidence; IF/ICC provides a localization readout in EFO-21 and U2OS, where supported staining is mainly vesicular with additional ER localization (HPA: subcellular ICC-IF).
HPA tissue IHC evidence for LMAN1
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 → |
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: LMAN1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced LMAN1 IHC Tips
Troubleshoot LMAN1 chromogenic IHC by checking retrieval, compartment-specific staining, controls and scoring; IF considerations are covered separately below.
Which retrieval condition should I start with for LMAN1 paraffin-section IHC?
Start with heat-mediated retrieval in EDTA, pH 8.0, the condition specified for this page’s catalog antibody (datasheet A03628-1). Keep heating and cooling times consistent across slides, then compare staining with a control section processed in the same run (standard IHC practice). If staining remains weak, adjust heating time in small increments while checking whether tissue morphology and background deteriorate (standard IHC practice). The supplied tissue image also used EDTA at pH 8.0, but its caption identifies antibody A03768-1, so that image does not establish performance for A03628-1 (caption: A03768-1; datasheet: A03628-1).
Could fixation explain weak or uneven LMAN1 staining?
The sensitivity of LMAN1 staining to fixation is unknown from the supplied target-specific evidence; the tissue-image caption identifies a paraffin section but does not state its fixative (caption: A03768-1). Record the fixative, fixation duration and processing history for each specimen before comparing staining intensity (standard IHC practice). On matched control sections, keep retrieval at EDTA, pH 8.0 while assessing whether the weak signal tracks a processing batch (datasheet A03628-1; standard IHC practice). Check morphology and staining at section edges, since uneven processing can complicate interpretation without establishing a LMAN1-specific fixation effect (standard IHC practice).
Where should convincing LMAN1 staining appear in chromogenic IHC?
Expect intracellular, predominantly cytoplasmic staining in tissue sections, consistent with the reported expression profile across tissues (HPA: cytoplasmic expression in all tissues). LMAN1 occupies ER–Golgi intermediate compartment, Golgi and ER membranes, while cell imaging places it mainly in vesicles with additional ER localisation (UniProt P49257 localisation; HPA: supported vesicles and ER). Assess signal within intact cells against the surrounding tissue rather than treating diffuse extracellular DAB as equivalent evidence (standard IHC practice). Strong isolated nuclear staining is inconsistent with those reported compartments and warrants review of counterstain, detection background and controls (UniProt P49257 localisation; standard IHC practice).
How do epitope position and processing affect LMAN1 IHC interpretation?
Check the antibody’s stated immunogen before selecting permeabilisation or interpreting loss of staining: LMAN1 has a lumenal region at residues 31–477, one transmembrane segment at 478–498 and a cytoplasmic tail at 499–510 (UniProt P49257 topology). Its signal peptide occupies residues 1–30, so an epitope there would require particular care when interpreting mature-protein staining (UniProt P49257 processing; standard IHC practice). The supplied record lists 0 isoforms and 0 glycosylation sites; do not invoke an undocumented splice variant or glycosylation site to explain discrepant staining (UniProt P49257). If epitope location is unavailable, compare staining with an independently validated antibody and matched controls before assigning a molecular cause (standard IHC practice).
How should I evaluate LMAN1 in a secondary IF experiment?
For IF multiplexing, pair LMAN1 with a marker identifying the cell population under study, and assess whether intracellular signal belongs to that population (standard IF practice). Glandular or endocrine cells in several tissues show high tissue-IHC staining, while cell imaging reports mainly vesicular signal with additional ER localisation (HPA: tissue IHC; HPA: supported vesicles and ER). Choose fluorophores after inspecting tissue autofluorescence, favouring a spectrally separated longer-wavelength channel when background is high (standard IF practice). Optimise permeabilisation against the antibody’s mapped epitope: residues 31–477 are lumenal, whereas 499–510 form the cytoplasmic tail (UniProt P49257 topology; standard IF practice).
How can I reduce misleading DAB background in LMAN1 sections?
Include a no-primary control and inspect whether brown deposits persist after omitting the catalog antibody; persistence implicates detection or tissue background (standard IHC practice). Apply an endogenous peroxidase block before HRP detection and titrate primary antibody and detection exposure against a consistently processed control (standard IHC practice). The supplied image used goat-serum blocking and DAB development, but its caption names A03768-1, so those observations cannot establish A03628-1-specific working conditions (caption: A03768-1). Recheck areas with edge staining, folds or damaged tissue before scoring, and require a cellular pattern compatible with intracellular LMAN1 localisation (standard IHC practice; UniProt P49257 localisation).
What is a defensible way to score LMAN1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and cell population to score before viewing group labels, then record the percentage of positive intact cells and an intensity-based H-score using the same thresholds across slides (standard IHC practice). Normalise each result to the number of evaluable cells in that population, and report the sampled area when using positive-cell density per mm² (standard IHC practice). Keep retrieval, staining run, DAB development and imaging settings consistent for comparisons (standard IHC practice). Because tissue-IHC expression is broadly cytoplasmic and RNA tissue specificity is low, document the cell population rather than relying on whole-section darkness alone (HPA: tissue profile and RNA specificity; standard IHC practice).
When should I doubt an apparent LMAN1-positive IHC result?
Prioritise staining within intact cells in an intracellular pattern: LMAN1 is reported in ER–Golgi intermediate compartment, Golgi and ER membranes, with vesicular and ER signal in cell imaging (UniProt P49257 localisation; HPA: supported vesicles and ER). A strong nuclear-only, extracellular or tissue-edge pattern conflicts with that localisation and should prompt review of morphology and controls (UniProt P49257 localisation; standard IHC practice). Exclude necrotic regions and compare with a no-primary control to identify deposits or endogenous enzyme signal that can mimic DAB positivity (standard IHC practice). HPA reports low staining in some cell populations despite cytoplasmic expression across tissues, so a faint signal alone is not proof of assay failure (HPA: tissue profile and low-staining populations).
Boster reagents
Best LMAN1 / Protein ERGIC-53 IHC Antibodies
Catalog antibodies list IHC and IF/ICC for human, mouse, and rat (catalog: applications/reactivity); supplied IF images show A549 cells and human paraffin tissue (A03628-1 IF captions).
Real IHC data
Anti-LMAN1 Antibody ®
Cat # A03628-1
A03628-1 is listed for IHC and IF/ICC in human, mouse, and rat (catalog: applications/reactivity), with IF images of A549 cells and human paraffin placenta and ovarian cancer sections (A03628-1 IF captions). M03628-1 and M03628-3 are rabbit monoclonals listed for those applications and species (catalog: host/clone/applications/reactivity), but neither has a supplied IHC or IF image (catalog: image alts).
Which to pick: For tissue IHC, A03628-1 lists 2–5 μg/ml (catalog: IHC dilution); its IHC captions describe human paraffin lung cancer and spleen sections but identify a different SKU, so confirm the figure attribution before relying on those images (IHC captions). The captions report no fixative (IHC captions). For IF/ICC, A03628-1 has direct image evidence (A03628-1 IF captions); M03628-1 and M03628-3 are rabbit monoclonal alternatives listed for human, mouse, and rat, without supplied images demonstrating cross-species staining (catalog: host/clone/reactivity/applications/image alts).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P49257 (LMAN1_HUMAN, Protein ERGIC-53).
- Human Protein Atlas. LMAN1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. LMAN1 subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the endoplasmic reticulum..
- Human Protein Atlas. LMAN1 antibody validation summary (2 antibodies).
- A dual role for ER-Golgi cargo receptor LMAN1 in supporting CSFV replication and restraining RLR signaling. Journal of virology 2026 — PMC13011374.
- Deciphering the YY1/ lncRNA BLACAT1/miR-605-3p axis in glioblastoma: implications for therapy. NPJ precision oncology 2025 — PMC12215738.
- PubMed PMID:8223692 — UniProt-cited evidence.
- PubMed PMID:7876089 — UniProt-cited evidence.
- PubMed PMID:10090935 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LMAN1 (Protein ERGIC-53). Expected localisation Tissue cytoplasm (HPA tissue IHC); ER/ERGIC/Golgi membranes (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03628-1). Validated IHC antibody: A03628-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A03628-1); 2. Fixation: Image fixative and duration unreported (datasheet A03628-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03628-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03628-1); 8. Primary antibody: Rabbit anti-LMAN1, 2-5μg/ml (datasheet A03628-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03628-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03628-1); Caveat: Low staining in some squamous epithelia despite broad expression (HPA tissue IHC); Isoform / epitope: No isoforms listed; cleaved signal; luminal vs cytoplasmic epitopes matter (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt P49257, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the…, Datasheet A03628-1, PMC13011374. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LMAN1/immunohistochemistry-ihc-guide.