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LMAN1 / Protein ERGIC-53 · IHC design guide
Design Immunohistochemistry for LMAN1
Plan LMAN1 staining in paraffin sections around its observed cytoplasmic tissue pattern (HPA tissue IHC) and ER–Golgi membrane location (UniProt). Start with the catalog antibody’s 2–5 μg/ml IHC range (datasheet A03628-1), and compare staining among the cell types reported by HPA (HPA tissue IHC).
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 | Cytoplasmic staining in tissue (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic; high in listed glandular/endocrine cells (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) | |
| Caveat | Signal may be low in smooth muscle cells (HPA tissue IHC) | |
| Regulation | Ubiquitous expression; regulation unreported (UniProt) | |
| Isoform / epitope | No isoforms; lumenal versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended LMAN1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A03628-1) is accompanied by one published paraffin-section protocol for LMAN1 staining (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-mediated retrieval (datasheet: A03628-1); the published protocol reports antigen unmasking without specifying its conditions (PMC13011374).
Section 2
What Is the Expected LMAN1 Staining Pattern?
LMAN1 is a membrane protein of the ER–Golgi intermediate compartment, Golgi and ER, with a lumenal region and one transmembrane segment (UniProt P49257 topology). In paraffin IHC, expect cytoplasmic staining across tissues, including strong staining in several glandular and endocrine cell populations (HPA tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in adrenal gland or breast glandular cells, or appendix endocrine cells (HPA tissue IHC). | This fits HPA's High ratings in those cells and its broader cytoplasmic profile (HPA tissue IHC). A granular or perinuclear accent would be consistent with ER–Golgi localisation (UniProt P49257); HPA tissue IHC does not require that finer pattern. |
| Predominantly nuclear or sharply cell-surface staining, with little cytoplasmic signal. | Those compartments conflict with the reported intracellular membrane locations (UniProt P49257; HPA tissue IHC). Treat the result as suspect and compare it with controls before assigning it to LMAN1; staining alone cannot identify the artefact. |
| Strong staining confined to an unexpected cell population while an expected High population is unstained (HPA tissue IHC). | Consider cross-reactivity or endogenous detection activity (general IHC practice). Because LMAN1 is ubiquitous (UniProt P49257), staining in another cell type alone is insufficient evidence of an error; the missing expected pattern makes the result concerning. |
| Uniform colour across tissue and empty areas, with cell boundaries hard to distinguish. | This does not resemble the cellular cytoplasmic profile reported for LMAN1 (HPA tissue IHC). Review background and detection controls; nonspecific reagent binding or residual detection activity are possible general IHC causes, not established LMAN1 properties. |
| No staining in a section containing an expected High population, such as gallbladder glandular cells (HPA tissue IHC). | An absent positive-control signal makes a negative study section difficult to interpret (general IHC practice). Check section quality and the staining workflow before concluding LMAN1 is absent; HPA's High rating does not diagnose which step failed. |
💡Expected LMAN1 appearanceCall positive a cellular cytoplasmic signal, strong in an HPA High glandular or endocrine population; isolated nuclear staining or colour outside cells is suspect (HPA tissue IHC; UniProt P49257; general IHC practice).
| Intracellular location and topology (UniProt P49257). | LMAN1 has a lumenal region at residues 31–477 and a transmembrane segment at 478–498 (UniProt P49257). These support interpreting cytoplasmic organelle staining, but do not establish how fixation or retrieval affects any antibody epitope. |
| Tissue contrast (HPA tissue IHC). | HPA rates several glandular and endocrine populations High, while smooth muscle cells and selected squamous epithelia are Low (HPA tissue IHC). Low means a weaker reference pattern here, not a proven negative control. |
| Antibody evidence (HPA antibody validation). | HPA lists HPA002320 and CAB037163 as Supported for IHC (HPA antibody validation). That supports their reported staining patterns, while the overall tissue profile has only medium antibody–RNA consistency (HPA tissue IHC); judge a new stain against controls. |
| IF/ICC Q: What localisation should a separate IF/ICC study show? | A: HPA reports mainly vesicles, with additional ER localisation, in ICC-IF (HPA subcellular). This is useful context for cytoplasmic IHC staining, but fluorescence detail should not be imposed as a required chromogenic paraffin-section pattern. |
| Signal-peptide processing (UniProt P49257). | The signal peptide spans residues 1–30; the annotated protein chain starts at 31 (UniProt P49257). Epitope location could matter for an antibody, but no epitope is supplied here, so no antibody-specific staining effect follows. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected High cells stain weakly throughout the section (HPA tissue IHC). | An underperforming staining workflow is possible (general IHC practice); no LMAN1-specific fixation sensitivity is reported here. | Check the IHC-validated antibody instructions and a known-positive section; review retrieval and detection settings as general IHC variables. |
| Colour appears in the detection-only control. | Endogenous detection activity or detection-reagent background is possible (general IHC practice). | Address the control signal before scoring LMAN1; review the relevant blocking and detection steps (general IHC practice). |
| Nuclei dominate while cytoplasm is faint. | The pattern disagrees with HPA's cytoplasmic tissue profile and UniProt's intracellular membrane locations (HPA tissue IHC; UniProt P49257). | Compare with a known-positive section and controls; reassess antibody specificity and staining conditions before calling it positive. |
| Every cell has similar heavy colour, obscuring tissue structure. | Excess background or overdeveloped chromogen may obscure cellular differences (general IHC practice). | Review antibody concentration, washes and chromogen development under the antibody's IHC instructions (general IHC practice). |
| A Low reference population stains more strongly than an expected High population (HPA tissue IHC). | The result diverges from HPA's relative tissue ratings, although HPA reports medium antibody–RNA consistency (HPA tissue IHC). | Confirm cell identity and compare matched sections, controls and antibody validation before interpreting the reversal. |
| No cells stain in the test section or known-positive section. | A shared workflow failure is possible (general IHC practice); this result cannot establish biological absence. | Check antibody application, retrieval, detection reagents and control-section integrity (general IHC practice); repeat with an HPA High population as the positive reference (HPA tissue IHC). |
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 breast first and score its glandular cells, which show High LMAN1 staining (HPA: Breast, 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 treat cells without staining on the positive slide as a background reference rather than confirmed antigen-negative cells (HPA: no negative rows; UniProt P49257: ubiquitous).
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: Run secondary-only and matched rabbit IgG isotype controls, matching the primary antibody’s clonality where applicable; a validated LMAN1 knockout is the preferred biological negative (caption: rabbit anti-LMAN1 primary). Quench endogenous peroxidase for chromogenic detection and compare any breast-tissue background with the no-primary slide (caption: peroxidase secondary and DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected tissue-IHC caption does not state a fixative; a paraffin section alone does not establish fixation (selected tissue-IHC caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used in the caption, but its antibody code A03768-1 differs from the selected SKU A03628-1, so retrieval dependence for the selected SKU is unestablished (selected tissue-IHC caption). Frozen-section performance and relative ease are unreported; ICC-IF images exist for EFO-21 and U2OS, with mainly vesicular and additional ER localization (HPA: subcellular images and supported locations). The selected A03628-1 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A03628-1).
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 staining in paraffin-section IHC by checking retrieval, compartment pattern, cell identity, controls and scoring before interpreting intensity.
Which retrieval should I try first for weak LMAN1 staining in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin-section IHC (datasheet A03628-1). Keep retrieval conditions identical across comparison slides, then assess whether cytoplasmic staining gains definition without an increase in tissue-wide background (standard IHC practice; HPA: cytoplasmic expression). The supplied tissue caption uses the same retrieval but names antibody A03768-1, while the selected SKU is A03628-1, so that image does not establish performance for the selected SKU (caption: A03768-1; selected SKU: A03628-1). If staining remains weak, compare a carefully titrated retrieval condition on serial sections and judge it against tissue integrity and control staining (standard IHC practice).
Could fixation explain weak or uneven LMAN1 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (caption: fixative not stated). Record the actual fixative and processing history for each block before attributing a weak signal to LMAN1 loss (standard IHC practice). Compare sections with similar processing and run them together using the specified EDTA retrieval at pH 8.0 (datasheet A03628-1; standard IHC practice). If staining differs between blocks, inspect morphology and control staining before changing retrieval or antibody concentration, because a processing difference alone does not establish a target-specific fixation effect (standard IHC practice).
What cellular pattern supports a specific LMAN1 IHC result?
Look for cytoplasmic staining with a perinuclear or punctate component, consistent with ER–Golgi intermediate compartment, Golgi and ER membrane localization (UniProt P49257: subcellular localization; HPA: cytoplasmic tissue expression). The HPA cell imaging summary places LMAN1 mainly in vesicles, with additional ER localization, but its microscopic pattern need not resolve identically in chromogenic tissue sections (HPA: subcellular summary; standard IHC practice). A diffuse nuclear-only pattern conflicts with the reported compartments and warrants review of controls and counterstain (UniProt P49257: subcellular localization; standard IHC practice). Compare cellular staining at the same magnification and exposure conditions across slides, documenting whether the expected cytoplasmic pattern persists (standard IHC practice).
How should epitope position affect my interpretation of LMAN1 staining?
Check the antibody's documented immunogen or epitope before interpreting an absent signal, because the supplied record does not identify the catalog antibody's binding site (record: epitope unspecified). Mature LMAN1 has a lumenal region at residues 31–477, a membrane span at 478–498 and a short cytoplasmic tail at 499–510 (UniProt P49257 topology). Its signal peptide occupies residues 1–30, and the record lists 0 isoforms and 0 glycosylation sites (UniProt P49257 processing, isoforms and glycosylation). If staining changes with retrieval, assess epitope accessibility and tissue preservation before proposing an isoform-specific or glycosylation-based explanation (standard IHC practice; UniProt P49257 record).
How can IF help check an ambiguous chromogenic LMAN1 pattern?
Use IF as an independent localization check while interpreting the paraffin-section IHC result against its own controls (standard IF/IHC practice). Multiplex LMAN1 with a marker for the expected cell population, such as a glandular-cell marker where those cells are being assessed, and confirm that both signals belong to the same cells (HPA: high glandular-cell staining in several tissues; standard IF practice). Choose a fluorophore away from prominent tissue autofluorescence and include a single-label control for each channel (standard IF practice). Because the antibody epitope is unspecified, optimize permeabilisation with its membrane side in mind: lumenal residues 31–477 and cytoplasmic residues 499–510 have different access requirements (record: epitope unspecified; UniProt P49257 topology; standard IF practice).
How do I reduce diffuse or misleading DAB background?
First compare the tissue section with a no-primary control and inspect whether DAB appears uniformly or only in plausible cellular compartments (standard IHC practice; UniProt P49257: subcellular localization). Include an endogenous peroxidase block when using HRP detection, and adjust blocking and washes if nonspecific color persists (standard IHC practice). The supplied caption reports 10% goat serum blocking and HRP/DAB detection for antibody A03768-1; those settings are method context, not matched-SKU validation for A03628-1 (caption: A03768-1; selected SKU: A03628-1). Titrate the IHC-validated antibody against a positive section, keeping DAB development comparable across slides before judging background reduction (standard IHC practice).
What is a defensible way to quantify LMAN1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, and exclude folds, necrosis and section edges using the same rules for every slide (standard IHC practice). For cellular staining, report percent positive cells and intensity grades 0–3; an H-score sums each grade times its percentage, giving a 0–300 range (standard IHC scoring practice). If counting stained cells per mm², normalize to evaluable tissue area and report the denominator rather than raw counts alone (standard IHC quantification practice). Keep retrieval, imaging and DAB development consistent, then compare like cell populations because LMAN1 staining intensity differs among reported tissue cell types (standard IHC practice; HPA: tissue IHC profile).
How can I distinguish genuine LMAN1 staining from artefact?
Favor reproducible cytoplasmic staining in intact cells, consistent with the reported ER–Golgi and vesicular locations, over nuclear-only color (UniProt P49257: subcellular localization; HPA: subcellular summary). Check whether the stained cell population matches the annotated region; HPA reports high staining in several glandular or endocrine populations and lower staining in several other listed populations (HPA: tissue IHC profile). Discount signal concentrated at section edges, in necrosis or in the no-primary control, and investigate endogenous peroxidase when HRP/DAB is used (standard IHC practice). HPA rates tissue staining as Supported with medium RNA–protein consistency, so resolve a discordant result with matched controls before inferring a biological change (HPA: reliability description; standard IHC practice).
Boster reagents
Best LMAN1 / Protein ERGIC-53 IHC Antibodies
The catalog antibody has human paraffin-section IHC images and human IF/ICC images; its listed reactivity includes Human, Mouse, and Rat (catalog: image captions, applications, reactivity).
Real IHC data
Anti-LMAN1 Antibody ®
Cat # A03628-1
A03628-1 is the SKU that will render; its IHC captions describe human lung cancer and spleen paraffin sections, but identify the antibody as A03768-1 (catalog: IHC captions). Its IF captions identify A03628-1 in A549 cells and human placenta and ovarian cancer paraffin sections (catalog: IF captions).
Which to pick: For tissue IHC, A03628-1 lists 2–5 μg/mL and EDTA pH 8 retrieval for human paraffin sections; confirm the antibody identity before relying on the figure because its IHC caption names A03768-1 (catalog: IHC dilution and caption). For IF/ICC, choose A03628-1 at the listed 5 μg/mL; its IF captions identify that SKU in both cells and tissue sections (catalog: applications, IF dilution and captions). For mouse or rat samples, A03628-1 lists reactivity but the supplied IHC/IF examples are human; the paraffin IHC caption does not report the fixative (catalog: reactivity and image captions).
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 Cytoplasmic staining in tissue (HPA tissue IHC). 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); Caveat: Signal may be low in smooth muscle cells (HPA tissue IHC); Isoform / epitope: No isoforms; lumenal versus cytoplasmic epitope matters (UniProt); Regulation: Ubiquitous expression; regulation unreported (UniProt). 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.