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LIAS / Lipoyl synthase, mitochondrial · IHC design guide
Design Immunohistochemistry for LIAS
Plan chromogenic LIAS IHC in paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC). Compare stained cell populations with controls, bearing in mind the low consistency between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
LIAS 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); mitochondrial LIAS (UniProt) | |
| Staining pattern | Cytoplasmic signal in glandular, neuronal and tubular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08974-1) | |
| Positive control | Cerebellum+4 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 A08974-1) | |
| Caveat | Staining shows low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; mature chain spans residues 28–372 (UniProt) |
Section 1
Recommended LIAS IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published LIAS IHC method for cartilage and knee-joint sections (PMC12283560).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08974-1) |
| Fixation | Image fixative and duration unreported (datasheet A08974-1); 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 A08974-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08974-1) |
| Primary antibody | Rabbit anti-LIAS, 1:50 recommended; image 1:100 (datasheet A08974-1) |
| Primary incubation | Overnight at 4 °C (datasheet A08974-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08974-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | LIAS-positive staining in purkinje cells of cerebellum (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A08974-1); the published method used EDTA pH 9.0 or citric acid pH 6.0 (PMC12283560).
Section 2
What Is the Expected LIAS Staining Pattern?
LIAS is a mitochondrial protein with no transmembrane segment (UniProt O43766 topology). In paraffin IHC, expect cytoplasmic staining in several tissues, including Purkinje cells, neurons, glandular cells, and kidney tubule cells (HPA tissue IHC). HPA rates the tissue staining Approved but reports low consistency with RNA expression and pending external verification (HPA tissue IHC). Interpret the pattern as provisional, especially when comparing tissues.
| Cytoplasmic staining in Purkinje cells, cortical neurons, colon glands, or kidney tubules. | These cells show Medium staining in HPA tissue IHC. Cytoplasmic chromogen agrees with HPA’s tissue profile and LIAS’s mitochondrial location (UniProt O43766 subcellular location). Routine chromogenic IHC cannot resolve individual mitochondria reliably (general IHC practice). |
| Predominantly nuclear staining in a paraffin section, with little cytoplasmic signal. | This differs from the HPA tissue IHC profile and warrants an artefact check (HPA tissue IHC; general IHC practice). Nuclear staining is not categorically impossible: HPA ICC-IF reports approved nucleoplasmic localisation as its main signal, with additional mitochondrial localisation (HPA subcellular ICC-IF). |
| Strong staining in adipocytes or esophageal squamous epithelium, especially across many cells. | HPA lists those cell types as Not detected in tissue IHC (HPA tissue IHC). Check for antibody cross-reactivity or endogenous chromogenic activity before assigning the signal to LIAS (general IHC practice). A negative HPA result is a comparison point, not proof of biological absence. |
| Weak, even colour over cells and tissue spaces, without a clear cellular pattern. | This does not resemble the reported cytoplasmic pattern (HPA tissue IHC). Diffuse colour may reflect nonspecific antibody binding, inadequate blocking, or detection background (general IHC practice); it cannot establish LIAS distribution. |
| No staining in an adequately preserved section containing HPA-positive cells. | Purkinje cells, colon glandular cells, and kidney tubule cells are reported at Medium intensity (HPA tissue IHC). First check the assay’s positive control, antibody conditions, retrieval, and detection steps (general IHC practice). HPA’s reported low RNA–staining consistency limits how strongly one negative section can be interpreted (HPA tissue IHC). |
💡Expected LIAS appearanceCall a result consistent with HPA when Medium cytoplasmic staining appears in its reported positive cells; broad staining of HPA Not detected cells or diffuse background needs investigation (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | HPA reports Medium staining in selected neurons, glandular cells, and kidney tubule cells, but Not detected staining in specified adipocytes, hematopoietic cells, and squamous cells (HPA tissue IHC). Compare the named cell population within a tissue, rather than treating a whole organ as uniformly positive or negative. |
| Compartment evidence across assays | UniProt places LIAS in mitochondria; HPA tissue IHC describes cytoplasmic expression, while approved HPA ICC-IF shows mainly nucleoplasm and additional mitochondria (UniProt O43766 subcellular location; HPA tissue IHC; HPA subcellular ICC-IF). This difference calls for assay-specific interpretation of a nuclear result. |
| Antibody and tissue evidence | Both listed antibodies, HPA018842 and HPA019076, have Approved IHC status; HPA nevertheless reports low consistency between staining and RNA expression and says external verification is pending (HPA antibodies; HPA tissue IHC). Approved status supports using the images as a guide, not treating every stain as confirmed LIAS. |
| Protein processing and variants | UniProt annotates a mature chain spanning residues 28–372 and 3 isoforms (UniProt O43766 processing; isoforms). No epitope position or isoform coverage is supplied, so those annotations cannot predict which variant the catalog antibody stains or justify a retrieval setting. |
| Retrieval conditions | No LIAS-specific retrieval condition or fixation sensitivity is supplied in the UniProt and HPA records. If signal is weak, review retrieval against the antibody’s actual instructions and an on-slide control (general IHC practice); do not infer a target-specific fixation effect from HPA staining levels. |
| Situation | Likely cause | Next action |
|---|---|---|
| A reported positive cell population has no chromogen. | Possible assay failure or weak detection; HPA’s Medium result is a reference observation, not a guarantee for every specimen (HPA tissue IHC; general IHC practice). | Check an on-slide positive control, antibody preparation, retrieval, and detection sequence; compare the same cell population in the HPA tissue images (general IHC practice; HPA tissue IHC). |
| The slide shows mainly nuclear staining. | The result conflicts with HPA’s cytoplasmic tissue IHC profile, although HPA ICC-IF reports approved nucleoplasmic signal (HPA tissue IHC; HPA subcellular ICC-IF). | Review morphology and controls, then repeat or confirm with an independent antibody before assigning the nuclear IHC signal to LIAS (general IHC practice; HPA antibodies). |
| HPA Not detected cells stain strongly. | Cross-reactivity or endogenous detection activity may explain staining in adipocytes or squamous cells (HPA tissue IHC; general IHC practice). | Inspect no-primary and detection controls, assess endogenous activity, and compare adjacent HPA-positive cell populations processed in the same run (general IHC practice; HPA tissue IHC). |
| Brown colour is widespread and obscures cell boundaries. | Nonspecific binding or detection background can produce a pattern unlike HPA’s cellular cytoplasmic staining (general IHC practice; HPA tissue IHC). | Check blocking, washing, antibody concentration, and no-primary control; score LIAS only after cell-associated signal can be separated from background (general IHC practice). |
| Two sections give different staining intensities. | Cell composition, assay variation, or genuine variation may contribute; HPA also flags low staining–RNA consistency (general IHC practice; HPA tissue IHC). | Compare matched cell types and run controls before interpreting the difference as a LIAS change (general IHC practice). Record the cell population and compartment alongside intensity. |
| Q: Should an IF/ICC image showing nucleoplasmic signal make nuclear IHC staining expected? | A: HPA ICC-IF reports mainly approved nucleoplasmic and additional mitochondrial localisation, whereas its tissue IHC profile is cytoplasmic (HPA subcellular ICC-IF; HPA tissue IHC). | Treat IF/ICC as a separate assay and confirm a nuclear paraffin IHC result independently; the IF/ICC pattern alone does not validate that IHC signal (general IHC practice). |
Experiment evidence
IHC & IF images showing LIAS staining
A08974-1 · IHCIHC analysis of LIAS using anti-LIAS antibody (A08974-1). LIAS was detected in a paraffin-embedded section of human breast cancer tissue. He
A08974-1 · IHCIHC analysis of LIAS using anti-LIAS antibody (A08974-1). LIAS was detected in a paraffin-embedded section of human thyroid cancer tissue. H
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000121897-LIAS/tissue/cerebral+cortex#imid_5087620
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000121897-LIAS/tissue/colon#imid_5087546
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000121897-LIAS/tissue/liver#imid_5087539
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000121897-LIAS/tissue/kidney#imid_5087540
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018842; green = LIAS, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018842; green = LIAS, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for LIAS IHC & IF
🧪Run cerebellum first and look for staining in Purkinje cells (HPA: Medium in cerebellar Purkinje cells). Run adipose tissue as a biological negative, focusing on adipocytes (HPA: Not detected in adipocytes); on the cerebellar slide, use neighboring cells without specific signal as an internal background reference, without assuming a particular cell type is LIAS-negative.
Positive control tissue: Cerebellum (Purkinje cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show LIAS in A-431, SK-MEL-30, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched nonimmune rabbit IgG isotype control (selected IHC caption: rabbit anti-LIAS primary); use LIAS knockout tissue or cells, if available, as a target-specific negative. Quench endogenous peroxidase for chromogenic detection and assess neuronal pigment against the controls in cerebellum (standard IHC practice).
⚠️Feasibility: A LIAS-specific fixation window or fixation effect is unreported, and the selected A08974-1 paraffin-section caption does not state the fixative (selected IHC caption). That caption uses heat retrieval in EDTA at pH 8.0, but does not establish retrieval dependence; the supplied evidence does not establish whether frozen sections or IF would be easier (selected IHC caption). In cerebellum, neuronal pigment can complicate chromogenic interpretation, and lipofuscin can autofluoresce in IF (standard IHC/IF practice).
HPA tissue IHC evidence for LIAS
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LIAS IHC Tips
Use the catalog antibody’s tissue IHC conditions as a starting point, then assess compartment, cell type, and staining controls before scoring LIAS.
What retrieval should I try first for weak LIAS staining in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA, pH 8.0 (datasheet A08974-1). The selected paraffin-section example used this retrieval before 1:100 primary antibody overnight at 4°C, so keep those conditions together during the first comparison (datasheet A08974-1). If staining remains weak, compare a longer heating period or an alternative retrieval buffer on matched sections, recording tissue preservation and background; those changes are troubleshooting trials, not reported LIAS conditions (standard IHC practice). Include a positive tissue compartment and a no-primary control in each run so a retrieval change is judged by specific staining rather than stronger DAB alone (standard IHC practice).
How should I handle fixation when LIAS staining varies between tissue blocks?
LIAS-specific fixation sensitivity is unknown: the selected tissue caption identifies a paraffin-embedded section but does not state its fixative (datasheet A08974-1). Record the fixative, fixation duration, block age, and processing history for each specimen, then compare sections processed together using identical retrieval and detection (standard IHC practice). For a prospective series, use one consistent fixation schedule and assess morphology alongside signal; avoid attributing a weak result to LIAS-specific formalin sensitivity without a controlled comparison (standard IHC practice). Keep the documented EDTA, pH 8.0 retrieval and 1:100 overnight primary incubation constant while testing fixation-related variation (datasheet A08974-1).
Which subcellular pattern is credible for LIAS in chromogenic IHC?
Assess LIAS as a mitochondrial protein, while recognizing that chromogenic tissue IHC may resolve this only as granular cytoplasmic staining (UniProt O43766 subcellular location; HPA tissue IHC profile). HPA tissue IHC describes cytoplasmic expression, whereas its separate ICC/IF assessment reports approved nucleoplasmic and additional mitochondrial localization (HPA tissue IHC; HPA subcellular). Treat strong, exclusively nuclear DAB as a pattern requiring independent confirmation rather than assuming the ICC/IF result validates nuclear staining in paraffin sections (HPA subcellular; standard IHC practice). Compare the same cell types across sections, use a no-primary control, and inspect high magnification for pigment or precipitate before assigning a compartment (standard IHC practice).
Can this IHC stain distinguish LIAS isoforms or reveal an inaccessible epitope?
LIAS has 3 listed isoforms and a radical SAM core spanning residues 122–341 (UniProt O43766 isoforms and domains). The supplied tissue caption does not identify the antibody epitope, so its staining cannot establish which isoform contributes signal or whether an epitope lies inside that core (datasheet A08974-1). LIAS has no annotated transmembrane segment and has a processed chain spanning residues 28–372, but neither annotation establishes epitope accessibility after tissue processing (UniProt O43766 topology and processing). If isoform discrimination matters, obtain the immunogen sequence and compare it with isoform sequences before interpreting differences in chromogenic staining (standard IHC practice).
How can I check LIAS localization with multiplex IF after tissue IHC?
For a separate IF experiment, pair LIAS with a marker identifying the expected cells, such as an epithelial marker for colon glandular cells reported at medium intensity (HPA tissue IHC: colon glandular cells). Add a mitochondrial marker to assess spatial agreement, while keeping nucleoplasmic signal open to evaluation because HPA ICC/IF reports approved nucleoplasmic and additional mitochondrial localization (HPA subcellular; standard IF practice). Choose fluorophores after checking tissue autofluorescence and place the weaker signal in a cleaner spectral channel, with single-stain and no-primary controls (standard IF practice). LIAS has no transmembrane segment, but its antibody epitope is unspecified; pilot mild detergent permeabilisation and verify mitochondrial antigen access without assuming which side of a membrane contains the epitope (UniProt O43766 topology; datasheet A08974-1; standard IF practice).
What should I change when LIAS DAB staining appears widespread?
First compare no-primary and secondary-only controls, because diffuse DAB can arise from detection chemistry or endogenous peroxidase rather than LIAS binding (standard IHC practice). Apply a peroxidase block before HRP detection and check that the block, washes, and DAB development are consistent across sections (standard IHC practice). The selected example used 10% goat serum blocking, 1:100 primary overnight at 4°C, and an HRP-linked secondary for 30 minutes at 37°C (datasheet A08974-1). If controls are clean but tissue staining is excessive, titrate the primary and shorten chromogen development while checking whether cytoplasmic detail and morphology remain interpretable (HPA tissue IHC profile; standard IHC practice).
How should I score LIAS staining across samples with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since HPA reports cytoplasmic staining in several tissues and medium signal in specific cells, including kidney tubules and colon glands (HPA tissue IHC). Within that population, record the percentage of positive cells and intensity to calculate an H-score, or report positive-cell density per mm² when spatial abundance is the question (standard IHC practice). Normalize counts to the number or area of eligible cells, excluding empty spaces and visibly damaged regions, and apply identical thresholds and imaging settings across the cohort (standard IHC practice). Report cell composition and control performance alongside scores because HPA flags low agreement between antibody staining and RNA expression (HPA tissue IHC reliability description).
How do I distinguish a true LIAS-positive cell from a staining artefact?
A credible result should occur in intact cells with a reproducible compartment pattern; LIAS is mitochondrial by UniProt, while HPA tissue IHC reports cytoplasmic expression (UniProt O43766 subcellular location; HPA tissue IHC profile). Compare the stained cell type with the reference pattern: kidney tubular cells are reported at medium intensity, whereas adipocytes are reported as not detected (HPA tissue IHC). Treat staining restricted to section edges, necrotic areas, debris, or a no-primary control as suspect, and assess endogenous peroxidase when DAB appears broadly (standard IHC practice). Interpret exclusively nuclear chromogenic signal cautiously because the approved nucleoplasmic observation comes from HPA ICC/IF and tissue IHC has a low staining-to-RNA consistency warning (HPA subcellular; HPA tissue IHC reliability description).
Boster reagents
Best LIAS / Lipoyl synthase, mitochondrial IHC Antibodies
The IHC-validated anti-LIAS antibody has IHC images from paraffin sections of human breast and thyroid cancer and mouse and rat heart (A08974-1 IHC captions). No IF/ICC data are supplied (catalog applications).
Real IHC data
Anti-LIAS Antibody ®
Cat # A08974-1
A08974-1 will render with an IHC image of a human breast cancer paraffin section (A08974-1 IHC caption). Its additional IHC captions show human thyroid cancer and mouse and rat heart paraffin sections (A08974-1 IHC captions).
Which to pick: Choose A08974-1 for tissue IHC: it is a rabbit polyclonal antibody listed for IHC, with paraffin-section images and human, mouse and rat reactivity (catalog applications, dilution data and reactivity; A08974-1 IHC captions). The catalog lists 1:50 for IHC, while the pictured sections used 1:100 after EDTA retrieval at pH 8.0; their fixative is unreported (catalog IHC dilution; A08974-1 IHC captions). No IF/ICC-validated SKU is provided, so A08974-1 has no supported IF/ICC recommendation here (catalog applications and IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O43766 (LIAS_HUMAN, Lipoyl synthase, mitochondrial).
- Human Protein Atlas. LIAS tissue IHC expression (reliability: Approved).
- Human Protein Atlas. LIAS subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the mitochondria..
- Human Protein Atlas. LIAS antibody validation summary (2 antibodies).
- C5a/C5aR pathway blocking promoted CuS-mediated cancer therapy effect by inhibiting cuproptosis resistance. Journal for immunotherapy of cancer 2025 — PMC12161435.
- A broad cuproptosis landscape in inflammatory bowel disease. Frontiers in immunology 2022 — PMC9669451.
- Tanshinone IIA induces oxidative stress in trophoblast cells and enhances copper dependent death. Scientific reports 2025 — PMC12254329.
- Hypoxia, cuproptosis, and osteoarthritis: Unraveling the molecular crosstalk. Redox biology 2025 — PMC12283560.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for LIAS (Lipoyl synthase, mitochondrial). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); mitochondrial LIAS (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08974-1). Validated IHC antibody: A08974-1 at 1:50 recommended; image 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08974-1); 2. Fixation: Image fixative and duration unreported (datasheet A08974-1); 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 A08974-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08974-1); 8. Primary antibody: Rabbit anti-LIAS, 1:50 recommended; image 1:100 (datasheet A08974-1); 9. Primary incubation: Overnight at 4 °C (datasheet A08974-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A08974-1); Caveat: Staining shows low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; mature chain spans residues 28–372 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt O43766, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A08974-1, PMC12283560. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/LIAS/immunohistochemistry-ihc-guide.