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- Table of Contents
LIAS / Lipoyl synthase, mitochondrial · Western blot design guide
Design a Western Blot for LIAS
Real validated LIAS Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LIAS WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
LIAS Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~41.9 kDa | |
| Observed band | ~39 kDa | |
| Gel | 12% (catalog A08974-1) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Observed below predicted | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated LIAS Western Blot Protocols
The A08974-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2, human Raji, human K562 (catalog A08974-1) |
| Gel % | 12% (catalog A08974-1) |
| Load | 30 ug; reducing conditions (catalog A08974-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A08974-1) |
| Membrane | nitrocellulose membrane (catalog A08974-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A08974-1) |
| Primary antibody | A08974-1 · 1:1000 (catalog A08974-1) |
| Primary incubation | overnight at 4°C (catalog A08974-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A08974-1) |
| Secondary incubation | 1.5 hour at RT (catalog A08974-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A08974-1) |
| Detection | ECL (catalog A08974-1) |
Section 2
What Is the Expected LIAS Western Blot Band Size?
LIAS is predicted at 41.9 kDa and observed near 39 kDa in reducing blots; the cause of the difference is not established.
| Band at ~39 kDa | Empirical LIAS band in reducing whole-cell blots; confirm identity with appropriate controls |
| Band near 41.9 kDa | Near the UniProt predicted mass; identity still needs confirmation |
| Several bands at different positions | Could include isoforms 1, 2, and 3 if their migration differs; distinct bands are not established |
| Weak band in mitochondrial-poor lysate | LIAS is mitochondrial, so sample composition may limit its signal |
💡Expected LIAS appearanceUniProt predicts 41.9 kDa, while reducing whole-cell blots show a ~39 kDa LIAS band; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass of 41.9 kDa | Provides a reference; the empirical band is ~39 kDa, with no established cause for the difference |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Alternative splicing could change apparent size, but its migration is not supplied |
| Isoform 3 | Alternative splicing could change apparent size, but its migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mitochondrial LIAS may be scarce in the sampled lysate | Check sample loading and compare with a mitochondria-enriched preparation |
| Band higher than expected | No supplied LIAS feature establishes a higher-mass band | Check molecular-weight markers and confirm identity with knockdown or an independent antibody |
| Band lower than expected | The observed ~39 kDa band is below the 41.9 kDa prediction; its cause is unknown | Compare with the empirical ~39 kDa reference and verify identity by knockdown or an independent antibody |
| Multiple bands | Isoforms 1, 2, and 3 exist, but distinct migration is unestablished | Test band identity with isoform-specific evidence or LIAS knockdown |
| Weak or no signal | LIAS is mitochondrial, and its abundance may be low in the sample | Check loading and enrichment using a mitochondrial marker |
Experiment evidence
Western blot images showing LIAS band
A08974-1 · LIAS
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LIAS Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LIAS in Western blot, you can use cerebellum tissue, which HPA reports as positive.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: LIAS is mitochondrial, so mitochondrial enrichment may improve its signal in tissue lysate.
HPA tissue expression evidence for LIAS
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for LIAS, answered from its protein features.
How should LIAS band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could LIAS isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Isoform 3 lacks residues 203–245, and isoform 2 lacks residues 323–372, so they may differ in mass. These sequence differences do not establish which isoforms are expressed or detectable in your sample.
Check the antibody epitope against all three isoforms. An epitope within UniProt residues 203–245 is absent from isoform 3; one within residues 323–372 is absent from isoform 2. These are UniProt coordinates and may differ from antibody documentation numbering.
Check the antibody epitope against all three isoforms. An epitope within UniProt residues 203–245 is absent from isoform 3; one within residues 323–372 is absent from isoform 2. These are UniProt coordinates and may differ from antibody documentation numbering.
Should LIAS bands be interpreted as glycosylated or modified forms?
PTM · The supplied features list no glycosylation sites or modified residues. Do not assign an extra band or the 39 kDa apparent mass to a specific modification from these data alone.
Does this guide establish induction of LIAS?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for LIAS?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A08974-1 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should LIAS be quantified if multiple bands appear?
Quantitation · Choose a band whose identity is supported by controls, then quantify that band consistently across samples. Record whether the antibody epitope covers all three isoforms; the listed sequence differences could affect which isoforms it detects.
Why might LIAS appear near 39 kDa instead of 41.9 kDa?
Interpretation · The reported apparent band is about 39 kDa, while the predicted mass is 41.9 kDa. LIAS is mitochondrial and has a transit peptide keyword, but these features alone do not establish the cause of the difference. Use the reported band as a reference and verify its identity with an appropriate control.
LIAS is annotated as mitochondrial. Comparing a mitochondrial fraction with an appropriate control fraction may help assess whether a candidate band follows the expected localization. Localization alone does not prove band identity.
Compare their apparent sizes with the reported ~39 kDa band and consider the listed isoform deletions. Check antibody epitope coverage and whether bands follow LIAS mitochondrial localization. The supplied features do not establish that any unexpected band is a specific isoform.
LIAS is annotated as mitochondrial. Comparing a mitochondrial fraction with an appropriate control fraction may help assess whether a candidate band follows the expected localization. Localization alone does not prove band identity.
Compare their apparent sizes with the reported ~39 kDa band and consider the listed isoform deletions. Check antibody epitope coverage and whether bands follow LIAS mitochondrial localization. The supplied features do not establish that any unexpected band is a specific isoform.
Boster reagents
LIAS Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-LIAS Antibody Picoband®
Cat # A08974-1
The listed anti-LIAS rabbit polyclonal antibody has a Western blot image using human cell lysates and rat and mouse brain and liver lysates. The reported band is approximately 39 kDa, versus an expected 42 kDa. Evidence is limited to this product caption.
Which to pick: A08974-1 is the only listed option. It reports human, mouse, and rat reactivity and includes a Western blot image with samples from all three species. Use the caption’s tested samples and conditions to judge fit for your experiment.
Source: BosterBio LIAS gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43766.
- Human Protein Atlas. LIAS tissue expression.
- PMC10462841 — target-verified WB comparison
- PMC12011582 — target-verified WB comparison
- PMC12715190 — target-verified WB comparison