This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SUCLG1 / Succinate--CoA ligase [ADP/GDP-forming] subunit alpha, mitochondrial · Western blot design guide
Design a Western Blot for SUCLG1
Source-linked SUCLG1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SUCLG1 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
SUCLG1 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 | ~36.3 kDa | |
| Observed band | ~35 kDa | |
| Gel | 5–20% (catalog A06274-1) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Hippocampus (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SUCLG1 Western Blot Protocol Options
The A06274-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human 293T, human HepG2, rat liver (catalog A06274-1) |
| Gel % | 5–20% (catalog A06274-1) |
| Load | 30 ug; reducing conditions (catalog A06274-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06274-1) |
| Membrane | nitrocellulose membrane (catalog A06274-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06274-1) |
| Primary antibody | A06274-1 · 0.5 μg/mL (catalog A06274-1) |
| Primary incubation | overnight at 4°C (catalog A06274-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06274-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06274-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06274-1) |
| Detection | ECL (catalog A06274-1) |
Section 2
What Is the Expected SUCLG1 Western Blot Band Size?
SUCLG1 is predicted at 36.3 kDa and observed at ~35 kDa; the cause of the difference is not established.
| Band at ~35 kDa | Empirical SUCLG1 band in reducing whole-cell lysates |
| Band near 36.3 kDa | Near the predicted SUCLG1 mass; verify identity |
| Little or no band in whole-cell lysate | Mitochondrial SUCLG1 may be more readily detected in a mitochondrial fraction |
| Band below the main band | Possible processing of the annotated transit peptide or degradation; identity needs confirmation |
💡Expected SUCLG1 appearanceSUCLG1 has a predicted mass of 36.3 kDa and an empirical band at ~35 kDa in reducing whole-cell lysates; confirm band identity with appropriate controls, since the cause of the difference is not established.
| Predicted SUCLG1 mass | Places the full-length sequence near 36.3 kDa; the observed band is ~35 kDa |
| Mitochondrial transit peptide annotation | Processing could lower mass, but the cleavage site and resulting band size are not supplied |
| N6-acetyllysine sites | Modification is documented at several residues, but a visible size shift is not established |
| N6-succinyllysine sites | Modification is documented, but a visible size shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Mitochondrial SUCLG1 may be scarce in the sampled whole-cell lysate | Check a mitochondrial-enriched fraction and a positive-control lysate |
| Band lower than expected | Transit peptide processing or degradation is possible; neither explains the ~35 kDa band conclusively | Compare with the ~35 kDa reference band and check band identity |
| Multiple bands | Modified SUCLG1 or nonspecific antibody binding is possible; distinct forms are not established | Compare with a validated positive control and an SUCLG1 depletion control |
| Weak or no signal | Low SUCLG1 abundance in the sampled mitochondrial material is possible | Check loading and mitochondrial enrichment against a positive control |
| Fragments below expected size | Protein degradation may produce smaller immunoreactive bands | Use fresh, protease-inhibited lysate and confirm the main ~35 kDa band |
Experiment evidence
Western blot images showing SUCLG1 band
A06274-1 · SUCLG1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SUCLG1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SUCLG1 in Western blot, you can use adrenal gland lysate.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Hippocampus (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: SUCLG1 is mitochondrial, so a mitochondrial-enriched lysate may improve detection.
HPA tissue expression evidence for SUCLG1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | white matter cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SUCLG1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SUCLG1, answered from its protein features.
Does the beta subunit determine which SUCLG1 band appears?
Band shift · SUCLG1 forms a heterodimer with a beta subunit; different beta subunits determine nucleotide specificity. The supplied features do not show that beta-subunit choice changes SUCLG1's apparent band position.
Could SUCLG1 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands; check antibody specificity before assigning additional bands to SUCLG1.
Which SUCLG1 modifications matter when interpreting a blot?
PTM · In UniProt sequence numbering, acetyllysine is listed at positions 54, 57, 66, 81 and 105; succinyllysine at 57, 66 and 338. These sites establish possible modifications, not a visible band shift.
UniProt marks acetyllysine and succinyllysine as alternate modifications at each of positions 57 and 66. Interpret them as alternatives at each residue, rather than assuming both occupy the same site on one molecule.
UniProt marks acetyllysine and succinyllysine as alternate modifications at each of positions 57 and 66. Interpret them as alternatives at each residue, rather than assuming both occupy the same site on one molecule.
Does this guide establish induction of SUCLG1?
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 SUCLG1?
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 A06274-1 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 total SUCLG1 and modified SUCLG1 be quantified?
Quantitation · Decide which signal the antibody measures. A total-SUCLG1 measurement cannot by itself quantify acetylation or succinylation at the listed sites; a modification-specific signal requires its own specificity assessment. Keep UniProt sequence numbering explicit when reporting sites.
Why is the SUCLG1 band near 35 kDa instead of 36.3 kDa?
Interpretation · The supplied apparent band is about 35 kDa, versus a predicted mass of 36.3 kDa. UniProt places SUCLG1 in mitochondria and lists a transit peptide keyword, but supplies no cleavage position. These features do not establish the cause of the mass difference.
Compare it with the approximately 35 kDa observed band and assess antibody specificity. The single listed isoform and modification sites alone do not identify an additional band or establish a shift.
Compare it with the approximately 35 kDa observed band and assess antibody specificity. The single listed isoform and modification sites alone do not identify an additional band or establish a shift.
Boster reagents
SUCLG1 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-SUCLG1 Antibody Picoband®
Cat # A06274-1
The catalog reports one anti-SUCLG1 antibody, A06274-1, with reported human and rat reactivity. Its Western blot image shows a band near 35 kDa in three human cell lysates and rat liver lysate; the expected size is 36 kDa. No independent validation is supplied.
Which to pick: A06274-1 is the only listed option. Its WB image covers HeLa, 293T, HepG2, and rat liver lysates using 30 µg per lane and 0.5 µg/mL primary antibody. Those examples support those tested contexts, not every human or rat sample.
Source: BosterBio SUCLG1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P53597.
- Human Protein Atlas. SUCLG1 tissue expression.
- PMC11709107 — target-verified WB comparison
- PMC2928220 — target-verified WB comparison
- PMC5126846 — target-verified WB comparison