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- Table of Contents
ACADL / Long-chain specific acyl-CoA dehydrogenase, mitochondrial · Western blot design guide
Design a Western Blot for ACADL
Real validated ACADL Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ACADL 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
ACADL 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 | ~47.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Kidney (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 | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ACADL Western Blot Protocols
The A07206 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | SHSY5Y (A), mouse liver (B), mouse heart (C), rat spinal cord (D) whole cell lysates (catalog A07206) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A07206; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ACADL Western Blot Band Size?
ACADL has a predicted precursor mass of 47.7 kDa; transit-peptide processing and modifications could affect migration, but no empirical band or demonstrated shift is supplied.
| Band near 47.7 kDa | Consistent with the predicted ACADL precursor size; confirm identity with controls |
| Band below 47.7 kDa | Could reflect removal of the annotated mitochondrial transit peptide; mature mass is not supplied |
| Closely spaced bands | Could reflect processing or modification states; their separation is not established |
| High-mass band under non-denaturing conditions | Could reflect the annotated homotetramer; confirm with denaturing conditions |
💡Expected ACADL appearanceUniProt predicts a 47.7 kDa ACADL precursor, but no empirical Western-blot band is supplied; use band-identity controls because transit-peptide processing and modifications have no quantified migration effects here.
| Predicted precursor mass | Sets a 47.7 kDa reference, not a validated band position |
| Mitochondrial transit peptide | Its removal could make mature ACADL smaller; the cleaved length is unspecified |
| Lysine acetylation, including residue 42 | May alter migration, but no visible shift is established |
| Lysine succinylation at residues 66 and 81 | May alter migration, but no visible shift is established |
| Phosphoserine 54 | May alter migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The mitochondrial fraction may have been lost during preparation | Check mitochondrial recovery and a positive-control lysate |
| Band higher than expected | Incomplete denaturation could retain the annotated homotetramer | Repeat with denaturing conditions and confirm identity with an independent antibody |
| Band lower than expected | Mitochondrial transit-peptide removal could reduce mass | Confirm identity with an independent antibody; do not assign a mature mass from this evidence |
| Multiple bands | Processing or modification states are possible, but distinct migration is unproven | Compare mitochondrial and whole-cell samples and validate bands with an independent antibody |
| Weak or no signal | Mitochondrial ACADL may be poorly recovered | Check mitochondrial recovery and antibody performance with a positive control |
| Fragments below expected size | Protein degradation during sample preparation is possible | Prepare fresh samples with protease inhibitors and compare the band pattern |
Experiment evidence
Western blot images showing ACADL band
A07206 · ACADL
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ACADL Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ACADL in Western blot, you can use kidney tissue.
Positive control: Kidney (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: ACADL is mitochondrial, so mitochondria-enriched lysate may improve detection.
HPA tissue expression evidence for ACADL
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 |
|---|---|---|---|---|
| Kidney | collecting ducts | High | Protein (IHC) | HPA → |
| Lung | alveolar cells type II | High | Protein (IHC) | HPA → |
| Thyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
| Liver | hepatocytes | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ACADL Western Blot Tips
Deeper troubleshooting and optimisation questions for ACADL, answered from its protein features.
How should ACADL 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 multiple ACADL bands represent different isoforms?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not support assigning additional bands to known ACADL isoforms.
Which ACADL modifications matter when interpreting bands?
PTM · UniProt lists acetyllysine at 42, 66, 81, 92, 95, 254, 279, 318, 322 and 358; succinyllysine at 66, 81, 165, 240, 254, 279 and 322; and phosphoserine at 54 and 362. These are UniProt sequence coordinates, which may differ from mature-protein or antibody numbering. Their presence alone does not establish a visible band shift.
UniProt lists them as alternate modifications at lysines 66, 81, 254, 279 and 322. Interpret these as alternative site assignments, not evidence that both modifications occupy the same residue on one molecule.
UniProt lists them as alternate modifications at lysines 66, 81, 254, 279 and 322. Interpret these as alternative site assignments, not evidence that both modifications occupy the same residue on one molecule.
Does this guide establish induction of ACADL?
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.
What transfer method to use for ACADL Western blot?
Transfer · ACADL is predicted to be 47.7 kDa. Check transfer around that size with a molecular-weight marker. The supplied features do not establish a specific membrane, transfer mode or transfer duration.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A07206 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 ACADL be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might ACADL migrate away from its predicted 47.7 kDa?
Interpretation · ACADL has a predicted mass of 47.7 kDa and a transit-peptide annotation, but no cleavage position or observed band size is supplied. A different apparent mass cannot be assigned to processing from these features alone. Compare the band with a molecular-weight marker without assuming an exact mature-protein mass.
ACADL is annotated in the mitochondrial matrix. Keep sample preparation and the amount loaded consistent across samples, especially when comparing mitochondrial fractions. Quantify the same identified band in each lane; the homotetramer annotation alone does not identify additional bands.
Check whether they occur near the predicted 47.7 kDa region and whether sample preparation could affect mitochondrial protein recovery. The listed modifications, transit-peptide keyword and homotetramer annotation do not, by themselves, establish the identity or apparent mass of another band.
ACADL is annotated in the mitochondrial matrix. Keep sample preparation and the amount loaded consistent across samples, especially when comparing mitochondrial fractions. Quantify the same identified band in each lane; the homotetramer annotation alone does not identify additional bands.
Check whether they occur near the predicted 47.7 kDa region and whether sample preparation could affect mitochondrial protein recovery. The listed modifications, transit-peptide keyword and homotetramer annotation do not, by themselves, establish the identity or apparent mass of another band.
Boster reagents
ACADL 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-ACADL Antibody
Cat # A07206
the supplier’s Anti-ACADL antibody A07206 lists human, mouse, and rat reactivity. Its Western blot image shows SHSY5Y, mouse liver, mouse heart, and rat spinal cord whole cell lysates; these are the reported tested samples, not validation of every tissue.
Which to pick: A07206 is the only listed ACADL antibody and has a Western blot image. Check whether its reported samples match your planned experiment.
Source: BosterBio ACADL gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P28330.
- Human Protein Atlas. ACADL tissue expression.
- PMC12026206 — target-verified WB comparison
- PMC10894480 — target-verified WB comparison
- PMC8841782 — target-verified WB comparison