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LMOD1 / Leiomodin-1 · IHC design guide
Design Immunohistochemistry for LMOD1
Plan LMOD1 paraffin-section IHC around cytoplasmic staining in smooth muscle and myoepithelial cells (HPA tissue IHC). Start with the catalog antibody at 1:50–1:200 and score staining by cell type (datasheet: IHC dilution; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LMOD1 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 tissue staining (HPA tissue IHC) | |
| Staining pattern | Selective cytoplasmic staining in smooth muscle and myoepithelial cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+3 more · see all | |
| Negative control | Appendix+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Adrenal glandular and placental endothelial cells also stain (HPA tissue IHC) | |
| Regulation | Staining-linked regulation is not established (UniProt) | |
| Isoform / epitope | 2 isoforms; isoform-specific epitope coverage is unreported (UniProt; datasheet) |
Section 1
Recommended LMOD1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published LMOD1 IHC methods for brain and visceral tissues, C. elegans, and leiomyosarcoma (PMC10439352; PMC5594508; PMC4526352).
| Sample | Paraffin-embedded human colon tissue; fixative not specified (datasheet A08666) |
| Fixation | Image fixative and duration unreported (datasheet A08666); 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-LMOD1, 1:50-1:200 (datasheet A08666) |
| 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 | LMOD1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Highly selective cytoplasmic expression in smooth muscle and myoepithelial cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule).
Section 2
What Is the Expected LMOD1 Staining Pattern?
LMOD1 should stain chiefly in the cytoplasm of smooth muscle and myoepithelial cells (HPA tissue IHC: highly selective cytoplasmic profile). It is a cytoplasmic, actin-associated protein with no transmembrane segment (UniProt P29536: subcellular location and topology). HPA rates the tissue IHC pattern Enhanced, citing high consistency between antibody staining and RNA expression (HPA tissue IHC: reliability).
| Cytoplasmic staining in smooth muscle cells, with little staining in adjacent unrelated cells (HPA tissue IHC: smooth muscle, Medium; selective profile). | This fits the expected pattern. Breast myoepithelial cells are another supported positive population (HPA tissue IHC: breast myoepithelial cells, Medium). Judge the relevant cell population, not the whole section, as positive. |
| Predominantly nuclear or cell-surface staining, without a convincing cytoplasmic component. | Treat this as a localisation mismatch: UniProt places LMOD1 in the cytoplasm and cytoskeleton, and HPA describes cytoplasmic tissue staining (UniProt P29536: subcellular location; HPA tissue IHC: profile). Review the staining pattern before scoring it as LMOD1. |
| Strong staining in cells listed as undetected, such as bone marrow hematopoietic cells or bronchial respiratory epithelial cells (HPA tissue IHC: Not detected). | Check for cross-reactivity or endogenous detection activity (standard IHC practice). Cell identity matters: HPA also reports High staining in placental endothelial and adrenal glandular cells, so those populations are not automatically unexpected (HPA tissue IHC: positive cells). |
| Diffuse chromogen across tissue and background, with no clear cytoplasmic cell pattern. | A diffuse deposit does not establish LMOD1 expression. Assess blocking, detection background and chromogen development with appropriate controls (standard chromogenic IHC practice); compare against HPA’s selective cytoplasmic profile (HPA tissue IHC: profile). |
| No signal in a section containing identifiable smooth muscle cells. | A negative result in a supported positive cell population needs a control check (HPA tissue IHC: smooth muscle cells, Medium). Confirm that the cells are present and that the detection run worked before interpreting the absence as biological (standard IHC practice). |
💡Expected LMOD1 appearanceCall a result consistent with LMOD1 when smooth muscle or breast myoepithelial cells show discernible medium cytoplasmic staining (HPA tissue IHC: Medium); isolated nuclear staining or diffuse background is not the reported pattern (UniProt P29536: cytoplasmic location; HPA tissue IHC: profile).
| Cell population and intensity | HPA reports Medium staining in smooth muscle and breast myoepithelial cells, but High staining in placental endothelial and adrenal glandular cells (HPA tissue IHC: positive cells). Interpret intensity with the identified cell type; a single intensity threshold cannot describe every listed positive population. |
| Subcellular position | UniProt places LMOD1 in the cytoplasm, myofibril and cytoskeleton and notes colocalisation with actin filaments in sarcomeres (UniProt P29536: subcellular location). HPA’s tissue summary is cytoplasmic (HPA tissue IHC: profile). Neither source supports a nuclear or membrane-only IHC call. |
| Antibody validation | Three listed rabbit polyclonal antibodies have Enhanced IHC status: HPA028325, HPA028435 and HPA030097 (HPA antibodies: IHC Enhanced). This supports the reported tissue pattern; it does not establish that every reagent or staining run will reproduce it (HPA: Enhanced reliability; standard IHC practice). |
| Isoforms and modifications | UniProt lists 2 isoforms and four example phosphoserine sites (UniProt P29536: isoforms and modified residues). The supplied sources do not map an IHC antibody epitope or show an isoform- or phosphorylation-dependent staining difference, so do not assign one. |
| Situation | Likely cause | Next action |
|---|---|---|
| Smooth muscle cells are unstained in an otherwise interpretable section (HPA tissue IHC: smooth muscle, Medium). | The target population may be absent from the examined field, or the IHC run may have failed (standard IHC practice); the supplied sources do not identify an LMOD1-specific fixation sensitivity. | Locate smooth muscle on the section and inspect a suitable positive control and detection controls before scoring the sample as negative (standard IHC practice). |
| The entire section has diffuse brown staining instead of selective cytoplasmic signal (HPA tissue IHC: profile). | Nonspecific background or endogenous detection activity can obscure a cell-specific pattern (standard chromogenic IHC practice). HPA’s tissue profile does not identify which workflow step caused it. | Compare with omission and detection controls; review blocking, washes and chromogen development under the laboratory’s IHC workflow (standard IHC practice). |
| Only nuclei or cell borders stain strongly. | That distribution conflicts with the documented cytoplasmic location (UniProt P29536: subcellular location; HPA tissue IHC: profile). It may reflect nonspecific staining (standard IHC interpretation). | Check morphology and controls, then repeat interpretation using cytoplasmic staining in a supported positive cell population as the reference (HPA tissue IHC: smooth muscle or myoepithelial cells). |
| Bronchial respiratory epithelium or bone marrow hematopoietic cells stain strongly (HPA tissue IHC: Not detected). | Cross-reactivity or endogenous detection activity is plausible (standard IHC practice); HPA reports these specific cell populations as undetected (HPA tissue IHC: negative cells). | Confirm the stained cell identity and compare with detection controls and a supported positive population before attributing that signal to LMOD1 (standard IHC practice; HPA tissue IHC: positive cells). |
| The slide contains strong placental endothelial or adrenal glandular staining. | These are reported High positive populations, despite HPA’s overall selective smooth muscle and myoepithelial summary (HPA tissue IHC: positive cells and profile). | Score the identified endothelial or glandular cells against the tissue-specific HPA observations; do not label them cross-reactive solely because they are outside the summary profile (HPA tissue IHC: positive cells). |
| IF/ICC Q: What intracellular pattern should be expected? | HPA approves cytosol as the main ICC-IF location; HPA028435 has Supported ICC status (HPA subcellular: Cytosol approved; HPA antibodies: ICC Supported). | A: Look for cytosolic localisation when interpreting IF/ICC images (HPA subcellular: Cytosol approved). Use the separate IF/ICC guide for its workflow. |
Experiment evidence
IHC & IF images showing LMOD1 staining
A08666 · IHCImmunohistochemistry of paraffin-embedded human colon using LMOD1 antibody at dilution of 1:200 (40x lens).
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163431-LMOD1/tissue/cerebral+cortex#imid_7633208
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163431-LMOD1/tissue/colon#imid_7632774
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163431-LMOD1/tissue/liver#imid_7632775
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163431-LMOD1/tissue/kidney#imid_7632768
![HPA ICC-IF BJ [Human fibroblast] — LMOD1 (HPA028435)](https://images.proteinatlas.org/28435/1446_H1_1_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA028435; green = LMOD1, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — LMOD1 (HPA028435)](https://images.proteinatlas.org/28435/1446_H1_6_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA028435; green = LMOD1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LMOD1 IHC & IF
🧪Run placenta first: its endothelial cells should stain (HPA: High in placental endothelial cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in bone marrow hematopoietic cells); on the placental slide, cells judged morphologically negative should show counterstain without specific chromogen deposition (standard IHC interpretation).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LMOD1 in BJ [Human fibroblast], U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species-, immunoglobulin-class-, and concentration-matched isotype controls; use validated LMOD1 knockout material or immunizing-peptide competition, if available, as a biological specificity control (standard IHC practice). In placental sections, quench endogenous peroxidase and check blood-containing areas for background chromogen (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the paraffin-section colon image at 1:200 does not report its fixative (catalog antibody caption: A08666). That caption supports paraffin-section IHC, but the supplied evidence does not establish whether frozen-section IHC or IF is easier; ICC-IF images show cytosolic localization in BJ, U-251MG and U2OS cells (HPA: subcellular). For placental IF, assess autofluorescence with an unstained section before interpreting endothelial signal (standard IF practice; HPA: High in placental endothelial cells).
HPA tissue IHC evidence for LMOD1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Endothelial cells | High | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LMOD1 IHC Tips
Use cell type and cytoplasmic localisation to assess LMOD1 staining, with the paraffin colon image as the catalog antibody’s direct tissue example.
How should I optimize retrieval when LMOD1 staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0 using heat induced retrieval at 95–98 °C for 20 min (page retrieval rule: cytoplasmic antigen). Keep section thickness, cooling time and detection conditions constant while comparing retrieval runs (standard IHC practice). The catalog antibody has a paraffin embedded human colon image at 1:200, but its caption does not report fixation or retrieval conditions (catalog antibody A08666 caption). If staining remains weak, test a different retrieval buffer or duration on adjacent sections and include a known positive smooth muscle area (standard IHC practice; HPA: smooth muscle cell expression). Judge improvement by cellular pattern as well as intensity, because LMOD1 is cytoplasmic and associated with actin filaments (HPA: cytosol; UniProt P29536: actin colocalisation).
Could fixation explain weak or patchy LMOD1 staining?
Target specific fixation sensitivity is unknown: the paraffin colon caption does not state its fixative, and no LMOD1 fixation comparison is supplied (catalog antibody A08666 caption). Record fixative, fixation duration and processing history for each block before attributing a weak result to the antibody (standard IHC practice). Compare adjacent sections from similarly processed positive tissue using the same pH 6.0, 95–98 °C, 20 min retrieval and detection run (page retrieval rule; standard IHC practice). Include a smooth muscle region as a tissue reference and assess whether its cytoplasmic signal survives processing (HPA: smooth muscle cell expression and cytoplasmic profile). Differences across blocks can suggest a processing issue, but they do not establish an LMOD1 specific fixation effect (standard IHC interpretation).
What staining compartment should I expect for LMOD1 in tissue?
Expect predominantly cytoplasmic staining in the relevant cells (HPA: highly selective cytoplasmic expression in smooth muscle and myoepithelial cells). UniProt also places LMOD1 with cytoskeletal structures and actin filaments in sarcomeres, so a filament associated pattern can be plausible in suitable muscle cells (UniProt P29536: subcellular localisation). Evaluate staining within the cell boundaries rather than assigning a nearby vessel’s signal to adjacent epithelium (standard IHC interpretation; HPA: smooth muscle expression). Strong isolated nuclear or luminal staining should prompt review of controls and morphology because neither is the reported principal location (HPA: cytoplasmic profile; UniProt P29536: subcellular localisation). Compare serial sections and a counterstain when cell identity is ambiguous (standard IHC practice).
How can I assess whether the antibody detects both LMOD1 isoforms?
LMOD1 has 2 annotated isoforms, while the supplied catalog caption does not identify the antibody’s binding epitope (UniProt P29536: isoforms; catalog antibody A08666 caption). Obtain the immunogen or epitope sequence and align it against both isoforms before claiming equal detection (standard antibody validation practice). The protein has a WH2 domain at residues 574–593 and annotated phosphoserines, including residues 12 and 555 (UniProt P29536: domains and modified residues). Those annotations alone do not show that either feature affects this antibody’s staining (UniProt P29536: annotations; catalog antibody A08666 caption). If isoform coverage matters, corroborate the staining pattern with an independently characterised reagent or an appropriate expression control (standard IHC validation practice).
How should I assess LMOD1 by multiplex immunofluorescence?
For the separate IF workflow, pair LMOD1 with a marker that identifies smooth muscle or myoepithelial cells, its expected tissue cell types (HPA: tissue IHC profile). Choose spectrally separated fluorophores and consider a far red LMOD1 channel where tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because LMOD1 is cytosolic and has no transmembrane segment, use a mild permeabilisation step when the staining assay requires intracellular antibody access (HPA: cytosol; UniProt P29536: topology; standard IF practice). Include single stain and no primary controls to check bleed through and autofluorescence (standard IF practice). Confirm any overlap within individual cells rather than treating adjacent signals as colocalisation (standard IF interpretation).
How do I distinguish LMOD1 signal from chromogenic background?
Run a no primary control alongside stained sections to identify detection background under the same conditions (standard IHC practice). For peroxidase based detection, block endogenous peroxidase before applying the detection system, and interpret DAB deposits against the control (standard chromogenic IHC practice). Adjust blocking, antibody dilution and wash conditions one variable at a time if diffuse staining obscures cells (standard IHC practice). The catalog image used 1:200 in paraffin embedded human colon, which provides a documented starting dilution rather than a universal optimum (catalog antibody A08666 caption). Prioritise reproducible cytoplasmic staining in smooth muscle over diffuse colour across unrelated structures (HPA: cytoplasmic smooth muscle profile; standard IHC interpretation).
What is a defensible way to quantify LMOD1 IHC staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, with smooth muscle cells assessed separately from neighbouring cell types (HPA: highly selective smooth muscle expression; standard IHC practice). For cytoplasmic chromogenic signal, report the percentage of positive cells and an intensity based H score, or report positive area or cell density per mm² when architecture makes cell counting difficult (standard IHC quantification practice). Normalise each measurement to the eligible cell count or annotated tissue area, and keep thresholds fixed across slides (standard IHC quantification practice). Exclude folds, damaged edges and necrotic regions by prespecified rules (standard IHC practice). Record batch controls and use the same counterstain and image settings for comparisons (standard IHC quantification practice).
When is an LMOD1 positive result convincing rather than artefactual?
A convincing result shows repeatable cytoplasmic signal in morphologically identified smooth muscle or myoepithelial cells, with controls behaving as expected (HPA: selective cytoplasmic profile; standard IHC validation practice). Interpret an unusual cell population cautiously: HPA reports glandular cells as not detected in appendix and several other sampled tissues, while its overall profile is selective (HPA: tissue IHC data). Nuclear dominant staining conflicts with the reported cytosolic location and warrants review (HPA: approved cytosol localisation). Edge concentrated colour, necrotic deposits or colour remaining in a no primary control can indicate technical artefact or endogenous enzyme activity (standard chromogenic IHC interpretation). Verify disputed staining on a repeat section and correlate it with tissue morphology (standard IHC practice).
Boster reagents
Best LMOD1 / Leiomodin-1 IHC Antibodies
The catalog includes an anti-LMOD1 antibody with real IHC data from paraffin-embedded human colon (A08666 IHC image caption). No IF image is supplied (A08666 IF image list).
Real IHC data
Anti-Leiomodin-1 LMOD1 Antibody
Cat # A08666
A08666 is listed for human IHC (catalog applications/reactivity). Its IHC image shows paraffin-embedded human colon at 1:200 (A08666 IHC image caption).
Which to pick: Choose A08666 for human paraffin-section IHC: its own image documents that preparation, while the fixative is unreported (A08666 IHC image caption). No listed SKU has IF/ICC validation or an IF image, so there is no supported IF/ICC pick (catalog applications/IF image list). A08666 is rabbit-hosted with clonality unreported and human-only listed reactivity; the payload supports no cross-species pick (catalog host/clone/reactivity fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P29536 (LMOD1_HUMAN, Leiomodin-1).
- Human Protein Atlas. LMOD1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. LMOD1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. LMOD1 antibody validation summary (3 antibodies).
- LMOD1, an oncogene associated with Lauren classification, regulates the metastasis of gastric cancer cells through the FAK-AKT/mTOR pathway. BMC cancer 2022 — PMC9055720.
- Nodding syndrome is unlikely to be an autoimmune reaction to leiomodin-1 after infection by Onchocerca volvulus. Biochemistry and biophysics reports 2023 — PMC10439352.
- Distinct molecular subtypes of uterine leiomyosarcoma respond differently to chemotherapy treatment. BMC cancer 2017 — PMC5594508.
- Clinically Relevant Molecular Subtypes in Leiomyosarcoma. Clinical cancer research : an official journal of the American Association for Cancer Research 2015 — PMC4526352.
- PubMed PMID:2026759 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LMOD1 (Leiomodin-1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A08666 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Appendix. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon tissue; fixative not specified (datasheet A08666); 2. Fixation: Image fixative and duration unreported (datasheet A08666); 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-LMOD1, 1:50-1:200 (datasheet A08666); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Adrenal glandular and placental endothelial cells also stain (HPA tissue IHC); Isoform / epitope: 2 isoforms; isoform-specific epitope coverage is unreported (UniProt; datasheet); Regulation: Staining-linked regulation is not established (UniProt). Evidence: UniProt P29536, HPA tissue IHC · Enhanced, HPA ICC-IF · cytosol., Datasheet A08666, PMC10439352, PMC5594508. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LMOD1/immunohistochemistry-ihc-guide.