This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SUMO3 / Small ubiquitin-related modifier 3 · Western blot design guide
Design a Western Blot for SUMO3
Real validated SUMO3 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SUMO3 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
SUMO3 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 | ~11.6 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation + Cleaved | |
| Caveat | Conjugation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SUMO3 Western Blot Protocols
The A05193 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse liver tissue lysate (catalog A05193) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A05193 · 2 μg/mL (catalog A05193) |
| 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 SUMO3 Western Blot Band Size?
Full-length SUMO3 is predicted at 11.6 kDa; cleavage, isoforms, and conjugation could affect migration, but no empirical band position is supplied.
| Band near 11.6 kDa | consistent with the predicted full-length SUMO3 mass; confirm identity with controls |
| Band slightly below 11.6 kDa | could reflect cleavage of the 93–103 propeptide |
| Higher-mass bands | could represent SUMO3 covalently attached to other proteins |
| Several bands at different positions | could include SUMO3 conjugates or isoforms 1 and 2; their migration is not established |
💡Expected SUMO3 appearanceFull-length SUMO3 has a predicted mass of 11.6 kDa; propeptide cleavage, isoforms, and conjugation may change the pattern, so confirm band identity with appropriate controls.
| UniProt predicted full-length mass | 11.6 kDa provides the reference size, not a measured band |
| Propeptide at residues 93–103 | cleavage could yield a smaller mature form; its apparent mass is unmeasured |
| Isoforms 1 and 2 | may differ in size, but their masses and migration are not supplied |
| Covalent attachment to other proteins | SUMO3-containing conjugates can appear above the free-protein band |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | SUMO3 may be covalently attached to another protein | compare with a SUMO3-depleted control to check band identity |
| Band lower than expected | the 93–103 propeptide may have been cleaved | compare precursor and mature SUMO3 controls if available |
| Multiple bands | SUMO3 conjugates or isoforms 1 and 2 may contribute | use a SUMO3-depleted control to identify specific bands |
| Broad smear instead of sharp band | SUMO3 attachment to different proteins may produce a range of higher-mass species | check whether the smear diminishes in a SUMO3-depleted control |
| Weak or no signal | free SUMO3 may be scarce relative to its conjugated forms | inspect higher-mass regions and verify detection with a positive control |
Experiment evidence
Western blot images showing SUMO3 band
A05193 · SUMO3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SUMO3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SUMO3 in Western blot, you can use adipose tissue, which has high HPA expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for SUMO3
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SUMO3 Western Blot Tips
Deeper troubleshooting and optimisation questions for SUMO3, answered from its protein features.
How should SUMO3 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 SUMO3 isoforms produce different bands?
Isoforms · Two isoforms are listed. In isoform 2, residue 50 is part of an alternative sequence annotated as Q to QVRHLAPPQSLPVCALVLCVPGIPRARASRGWTQMQLPE. Consider isoform expression when assessing bands, but these features do not establish which isoform is present or where either migrates.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of SUMO3?
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 SUMO3?
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 A05193 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.
What should be quantified on a SUMO3 blot?
Quantitation · Decide whether the question concerns free SUMO3 or SUMO3 attached to other proteins, and quantify those signals separately. UniProt describes covalent attachment to multiple proteins and lists a propeptide at residues 93–103; a single band need not represent every SUMO3-containing form.
Why might SUMO3 migrate differently from its predicted 11.6 kDa mass?
Interpretation · The 11.6 kDa prediction describes the 103-residue sequence. UniProt lists a propeptide at residues 93–103 and covalent attachment to other proteins. These features give reasons to investigate a different apparent mass, but neither establishes where a band will migrate; no observed band mass is supplied.
UniProt marks residues 93–103 as a propeptide. Consider processing when comparing a band with the 11.6 kDa prediction for the 103-residue sequence. The supplied features do not give a measured mass or migration position for processed SUMO3.
UniProt describes SUMO3 as covalently attached to a number of proteins, so conjugated species are a possibility. The annotation alone cannot identify the attached protein or prove that a particular band is a SUMO3 conjugate.
SUMO3 is annotated in the cytoplasm, nucleus, and PML bodies. Compare fractions if band localization matters, while allowing for SUMO3 signal in both cytoplasmic and nuclear samples. Location alone does not assign a band to a specific conjugate or isoform.
UniProt marks residues 93–103 as a propeptide. Consider processing when comparing a band with the 11.6 kDa prediction for the 103-residue sequence. The supplied features do not give a measured mass or migration position for processed SUMO3.
UniProt describes SUMO3 as covalently attached to a number of proteins, so conjugated species are a possibility. The annotation alone cannot identify the attached protein or prove that a particular band is a SUMO3 conjugate.
SUMO3 is annotated in the cytoplasm, nucleus, and PML bodies. Compare fractions if band localization matters, while allowing for SUMO3 signal in both cytoplasmic and nuclear samples. Location alone does not assign a band to a specific conjugate or isoform.
Boster reagents
SUMO3 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-SUMO3 Antibody
Cat # A05193
A05193 is an anti-SUMO3 antibody listed as reactive with human, mouse, and rat. Its Western blot image shows mouse liver tissue lysate tested at 2 μg/mL; the supplied evidence does not show Western blot results for other samples.
Which to pick: A05193 is the only listed option. Its reported reactivity covers human, mouse, and rat, while the supplied Western blot example specifically uses mouse liver lysate at 2 μg/mL.
Source: BosterBio SUMO3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P55854.
- Human Protein Atlas. SUMO3 tissue expression.
- PMC3264615 — target-verified WB comparison
- PMC11345958 — target-verified WB comparison
- PMC5775435 — target-verified WB comparison