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- Table of Contents
SUMO2 / Small ubiquitin-related modifier 2 · Western blot design guide
Design a Western Blot for SUMO2
Real validated SUMO2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SUMO2 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
SUMO2 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 | ~10.9 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Adrenal gland (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 | Acetylated + Ubl conjugation | |
| Caveat | Conjugated species possible | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SUMO2 Western Blot Protocols
The A01282-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog A01282-1) |
| 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 | A01282-1 · (A) 1 and (B) 2 μg/mL (catalog A01282-1) |
| 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 SUMO2 Western Blot Band Size?
SUMO2 has a predicted 10.9 kDa precursor; processing, isoforms, and conjugation could affect bands, but their migration is not demonstrated here.
| Band near 10.9 kDa | Possible unconjugated SUMO2; 10.9 kDa is the predicted precursor mass. |
| Slightly lower band, if resolved | Could reflect removal of propeptide residues 94–95; the small change may not resolve. |
| Several low-mass bands | Could include isoforms 1 and 2, though distinct migration is not established. |
| Higher-mass bands or ladder | Could represent SUMO2 conjugated to other proteins through isopeptide bonds. |
💡Expected SUMO2 appearanceUniProt predicts a 10.9 kDa SUMO2 precursor, but supplies no empirical band; processing, isoforms, and conjugation may affect migration, so confirm band identity with antibody and conjugation controls.
| UniProt predicted precursor mass | Places unconjugated SUMO2 near 10.9 kDa before processing. |
| Propeptide residues 94–95 | Their removal slightly reduces mass, with no established resolvable shift. |
| Splice isoform 1 | Its size relative to isoform 2 is not supplied. |
| Splice isoform 2 | Its size relative to isoform 1 is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear enrichment may leave little detectable SUMO2 in the sampled lysate. | Check a nuclear fraction and a positive control. |
| Band higher than expected | SUMO2 may be covalently conjugated to another protein. | Compare with an unconjugated SUMO2 control and confirm SUMO2 identity. |
| Band lower than expected | Propeptide removal slightly lowers mass, but may not explain a clearly lower band. | Check antibody specificity and compare precursor and mature SUMO2 controls. |
| Broad smear instead of sharp band | SUMO2 conjugates with different protein partners may span many sizes. | Compare with a SUMO2 conjugation control and verify antibody specificity. |
| Multiple bands | Isoforms or SUMO2 conjugates may contribute; distinct isoform bands are unproven. | Check band identity with isoform-aware and conjugation controls. |
| Weak or no signal | SUMO2 is localized to the nucleus and PML bodies. | Check nuclear enrichment, sample loading, and an antibody positive control. |
Experiment evidence
Western blot images showing SUMO2 band
A01282-1 · SUMO2
M01282-2 · SUMO2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SUMO2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SUMO2 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for SUMO2
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 → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial 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 |
|---|---|---|---|---|
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Low | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SUMO2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SUMO2, answered from its protein features.
How should SUMO2 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 SUMO2 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 51..74 in UniProt numbering, so it may have a different mass. The supplied features do not establish whether both isoforms are expressed in your sample or resolve as separate bands.
Does SUMO2 acetylation explain a shifted band?
PTM · UniProt annotates alternate N6-acetyllysine at position 11, using UniProt sequence numbering. Check that numbering convention when comparing antibody or paper annotations. This modification alone does not establish a visible shift or explain a measured mass difference.
Does this guide establish induction of SUMO2?
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 SUMO2?
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 A01282-1 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 free and conjugated SUMO2 be quantified?
Quantitation · Define whether the measurement targets the putative free SUMO2 region or higher-mass SUMO2 signal, and report them separately. SUMO2 has conjugation-related annotations, so combining those regions could obscure which signal changed. The supplied features do not identify band boundaries or establish that every higher-mass band contains SUMO2.
Should SUMO2 run at its predicted 10.9 kDa?
Interpretation · 10.9 kDa is the predicted mass; no observed band position is supplied. UniProt annotates a propeptide at residues 94..95, but these features do not establish where a free SUMO2 band will migrate. Use the predicted mass as a starting reference, not a required band position.
UniProt annotates residues 94..95 as a propeptide on the 95-residue sequence. Consider that annotation when comparing a band with the 10.9 kDa predicted mass. No observed band position or processing measurement is supplied, so do not assign a band solely from this feature.
SUMO2 is annotated in a ubiquitin-like conjugation pathway, and UniProt describes interactions involving SUMO2 chains, ZNF451, and UBE2I/UBC9. Higher-mass signal could therefore warrant investigation as conjugated SUMO2. These annotations alone cannot identify a particular band or its conjugated protein.
Compare the band pattern with the 10.9 kDa predicted mass, the propeptide at UniProt residues 94..95, isoform 2's missing residues 51..74, and SUMO2 conjugation. UniProt also lists alternate acetylation at Lys11, but its presence does not prove a visible shift. Without an observed band position, none of these features assigns a specific band.
UniProt annotates residues 94..95 as a propeptide on the 95-residue sequence. Consider that annotation when comparing a band with the 10.9 kDa predicted mass. No observed band position or processing measurement is supplied, so do not assign a band solely from this feature.
SUMO2 is annotated in a ubiquitin-like conjugation pathway, and UniProt describes interactions involving SUMO2 chains, ZNF451, and UBE2I/UBC9. Higher-mass signal could therefore warrant investigation as conjugated SUMO2. These annotations alone cannot identify a particular band or its conjugated protein.
Compare the band pattern with the 10.9 kDa predicted mass, the propeptide at UniProt residues 94..95, isoform 2's missing residues 51..74, and SUMO2 conjugation. UniProt also lists alternate acetylation at Lys11, but its presence does not prove a visible shift. Without an observed band position, none of these features assigns a specific band.
Boster reagents
SUMO2 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-SUMO2/3 Antibody
Cat # A01282-1
Real WB data
Anti-Sumo2/3 Rabbit Monoclonal Antibody
Cat # M01282-2
The catalog reports two anti-SUMO2/3 antibodies, A01282-1 and M01282-2. Both list human, mouse, and rat reactivity and show Western blots of Jurkat cell lysate. The supplied evidence does not establish that either distinguishes SUMO2 from SUMO3.
Which to pick: Both have Jurkat lysate Western blot images and the same listed reactivity. Pick A01282-1 if you need a reported antibody concentration (1 or 2 μg/mL). M01282-2 is the rabbit monoclonal option; its caption does not report a concentration.
Source: BosterBio SUMO2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P61956.
- Human Protein Atlas. SUMO2 tissue expression.
- PMC13481645 — target-verified WB comparison
- PMC12931719 — target-verified WB comparison
- PMC4603762 — target-verified WB comparison