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- Table of Contents
SUCLA2 / Succinate--CoA ligase [ADP-forming] subunit beta, mitochondrial · Western blot design guide
Design a Western Blot for SUCLA2
Source-linked SUCLA2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SUCLA2 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
SUCLA2 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~50.3 kDa | |
| Observed band | Approximately 48 kDa | |
| Gel | 5–20% (catalog A04807-1) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Ovary (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 | 2 isoform(s) |
Section 1
Source-Linked SUCLA2 Western Blot Protocol Options
The A04807-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Caco-2, human 293T, human MOLT-4, human HepG2, rat lung, rat brain, mouse lung, mouse brain (catalog A04807-1) |
| Gel % | 5–20% (catalog A04807-1) |
| Load | 30 ug; reducing conditions (catalog A04807-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04807-1) |
| Membrane | nitrocellulose membrane (catalog A04807-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04807-1) |
| Primary antibody | A04807-1 · 0.25 μg/mL (catalog A04807-1) |
| Primary incubation | overnight at 4°C (catalog A04807-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04807-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04807-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04807-1) |
| Detection | ECL (catalog A04807-1) |
Section 2
What Is the Expected SUCLA2 Western Blot Band Size?
SUCLA2 is predicted at 50.3 kDa and observed near 48 kDa; the cause of the difference is not established.
| Band near 48 kDa | Empirical SUCLA2 band in reducing whole-cell lysates; confirm identity with antibody controls. |
| Band near 50.3 kDa | Compatible with the predicted full-length polypeptide mass, but identity requires confirmation. |
| Band below 50.3 kDa | Could reflect mitochondrial transit-peptide processing; the mature mass is not supplied. |
| Several nearby bands | Isoforms 1 and 2 exist, but their migration difference is not established. |
💡Expected SUCLA2 appearanceUniProt predicts 50.3 kDa, while antibody QC reports a band at approximately 48 kDa in reducing whole-cell lysates; confirm band identity with ordinary antibody controls because the cause of the difference is unproven.
| Predicted full-length mass | 50.3 kDa is the reference polypeptide mass; the empirical band is approximately 48 kDa. |
| Mitochondrial transit peptide | Processing could lower the mature protein mass, but no cleavage position or resulting mass is supplied. |
| Isoform 1 | Its mass and migration relative to isoform 2 are not supplied. |
| Isoform 2 | Its mass and migration relative to isoform 1 are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mitochondrial SUCLA2 may be insufficiently represented in the sample. | Check lysate quality and compare with a mitochondria-enriched sample. |
| Band higher than expected | The band may be nonspecific; no SUCLA2 size-increasing feature is established here. | Check antibody specificity with a SUCLA2 depletion control. |
| Band lower than expected | Transit-peptide processing is possible, but its contribution to migration is unknown. | Compare with the reported 48 kDa band and verify identity by SUCLA2 depletion. |
| Multiple bands | Isoforms 1 and 2 exist, but distinct bands are not demonstrated. | Use SUCLA2 depletion to identify which bands depend on SUCLA2. |
| Weak or no signal | Low SUCLA2 abundance or poor recovery from the mitochondrial fraction is possible. | Check sample loading and transfer, then compare with a mitochondria-enriched sample. |
Experiment evidence
Western blot images showing SUCLA2 band
A04807-1 · SUCLA2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SUCLA2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SUCLA2 in Western blot, you can use cerebellum tissue lysate, which HPA rates High.
Positive control: Cerebellum (IHC candidate; verify WB)
Negative control: Ovary (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: A tissue lysate is suitable for mitochondrial SUCLA2, and HPA lists ovary as not detected.
HPA tissue expression evidence for SUCLA2
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 | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Liver | hepatocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SUCLA2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SUCLA2, answered from its protein features.
How should SUCLA2 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 SUCLA2 isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 lacks residues 26–47 of the canonical sequence, so it differs in length from isoform 1. The supplied features do not establish whether the isoforms resolve as separate Western blot bands.
Decide whether the measured band represents one isoform or a combined signal. Isoform 2 lacks canonical residues 26–47, but the supplied features do not show whether the isoforms separate on the blot or whether an antibody detects both. Apply the same band-selection rule across samples.
Decide whether the measured band represents one isoform or a combined signal. Isoform 2 lacks canonical residues 26–47, but the supplied features do not show whether the isoforms separate on the blot or whether an antibody detects both. Apply the same band-selection rule across samples.
Which SUCLA2 lysine modifications should I consider?
PTM · The supplied UniProt coordinates list N6-acetyllysine at 78, 88, 129, 139, 143, 216 and 368. At 88, N6-succinyllysine is listed as an alternate modification. These entries identify possible modified residues; they do not establish a visible band shift. Check coordinate conventions before comparing them with antibody or paper numbering.
The supplied UniProt coordinates list phosphotyrosine at 84, phosphoserine at 279 and phosphothreonine at 341. Consider these sites when interpreting a phosphorylation-specific assay, but their presence alone does not establish a detectable shift in a total-SUCLA2 blot. Confirm the numbering convention before comparing sites across sources.
The supplied UniProt coordinates list phosphotyrosine at 84, phosphoserine at 279 and phosphothreonine at 341. Consider these sites when interpreting a phosphorylation-specific assay, but their presence alone does not establish a detectable shift in a total-SUCLA2 blot. Confirm the numbering convention before comparing sites across sources.
Does this guide establish induction of SUCLA2?
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 SUCLA2?
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 A04807-1 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 SUCLA2 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 is the SUCLA2 band near 48 kDa instead of 50.3 kDa?
Interpretation · The observed band is approximately 48 kDa, while the supplied predicted mass is 50.3 kDa. SUCLA2 is mitochondrial and has a transit peptide keyword, but no cleavage site or mature mass is supplied. These features do not establish the cause of the difference in apparent mass.
Consider the listed isoforms and modifications as possibilities, then verify band identity experimentally. Isoform 2 lacks residues 26–47; acetylation, succinylation and phosphorylation sites are listed. None of those feature entries proves that a particular unexpected band is SUCLA2 or explains its apparent mass.
SUCLA2 is listed as mitochondrial. For comparisons, use a consistent sample preparation and compare equivalent sample types, since a mitochondrial fraction and a whole-cell sample need not give comparable SUCLA2 signal. The supplied features do not specify an expected abundance or a normalization control.
Consider the listed isoforms and modifications as possibilities, then verify band identity experimentally. Isoform 2 lacks residues 26–47; acetylation, succinylation and phosphorylation sites are listed. None of those feature entries proves that a particular unexpected band is SUCLA2 or explains its apparent mass.
SUCLA2 is listed as mitochondrial. For comparisons, use a consistent sample preparation and compare equivalent sample types, since a mitochondrial fraction and a whole-cell sample need not give comparable SUCLA2 signal. The supplied features do not specify an expected abundance or a normalization control.
Boster reagents
SUCLA2 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-SUCLA2 Antibody Picoband®
Cat # A04807-1
A04807-1 is a rabbit polyclonal anti-SUCLA2 antibody with a Western blot image showing an approximately 48 kDa band in human cell lysates and rat and mouse tissue lysates. The supplied evidence does not include independent validation.
Which to pick: A04807-1 is the only listed option. Its WB image includes human Caco-2, 293T, MOLT-4, and HepG2 cells plus rat and mouse lung and brain tissue, tested with 30 µg of reducing lysate and 0.25 µg/mL primary antibody.
Source: BosterBio SUCLA2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9P2R7.
- Human Protein Atlas. SUCLA2 tissue expression.
- PMC5346465 — target-verified WB comparison
- PMC13519632 — target-verified WB comparison
- PMC5126846 — target-verified WB comparison