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- Table of Contents
UBE2S / Ubiquitin-conjugating enzyme E2 S · Western blot design guide
Design a Western Blot for UBE2S
Source-linked UBE2S Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBE2S 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
UBE2S 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 | ~23.8 kDa | |
| Observed band | ~24 kDa | |
| Gel | 5–20% (catalog A03045-2) | |
| Positive control | Appendix (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked UBE2S Western Blot Protocol Options
The A03045-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human Hela, human Jurkat, human MCF-7, rat testis (catalog A03045-2) |
| Gel % | 5–20% (catalog A03045-2) |
| Load | 30 ug; reducing conditions (catalog A03045-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03045-2) |
| Membrane | nitrocellulose membrane (catalog A03045-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03045-2) |
| Primary antibody | A03045-2 · 0.5 μg/mL (catalog A03045-2) |
| Primary incubation | overnight at 4°C (catalog A03045-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03045-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03045-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03045-2) |
| Detection | ECL (catalog A03045-2) |
Section 2
What Is the Expected UBE2S Western Blot Band Size?
UBE2S is predicted at 23.8 kDa and observed near 24 kDa; its listed modifications have no demonstrated effect on that migration.
| Band near 24 kDa | matches the empirical UBE2S band |
| Single sharp band near 24 kDa | consistent with one annotated UBE2S isoform |
| No separate acetylated band | Met1 acetylation need not produce a visible shift |
| No separate phosphorylated band | Ser173 phosphorylation need not produce a visible shift |
💡Expected UBE2S appearanceUBE2S has a predicted mass of 23.8 kDa and an observed band near 24 kDa; confirm band identity with an appropriate specificity control.
| Predicted UBE2S mass | 23.8 kDa, close to the observed ~24 kDa band |
| Met1 N-acetylation | a listed modification with no demonstrated visible shift |
| Ser173 phosphorylation | a listed modification with no demonstrated visible shift |
| APC/C complex association | does not establish a higher molecular weight UBE2S band on SDS-PAGE |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | identity or migration of the higher band is unestablished | compare with the ~24 kDa band and validate specificity |
| Band lower than expected | identity of the lower band is unestablished | check sample integrity and validate specificity |
| Multiple bands | the single annotated isoform does not establish multiple UBE2S sizes | identify which band tracks with UBE2S in a specificity control |
| Weak or no signal | UBE2S may be below the assay's detection limit | check transfer and include a known positive lysate |
| Fragments below expected size | sample degradation is possible; no cleavage product is annotated | check sample handling and validate fragment identity |
Experiment evidence
Western blot images showing UBE2S band
A03045-2 · UBE2S
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBE2S Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBE2S in Western blot, you can use appendix tissue, where HPA reports medium expression.
Positive control: Appendix (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: HPA reports UBE2S as not detected in adipose tissue, though the positive appendix signal is only medium.
HPA tissue expression evidence for UBE2S
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Medium | Protein (IHC) | HPA → |
| Testis | cells in seminiferous ducts | Medium | Protein (IHC) | HPA → |
| Tonsil | germinal center cells | Medium | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBE2S Western Blot Tips
Deeper troubleshooting and optimisation questions for UBE2S, answered from its protein features.
How should UBE2S 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.
Should UBE2S isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Could phosphorylation shift the UBE2S band?
PTM · UniProt lists phosphoserine at position 173 in its sequence numbering. Compare samples with an appropriate phosphorylation control if testing this possibility. The listed site alone does not establish a visible shift.
UniProt lists N-acetylmethionine at position 1 in its sequence numbering. This modification is documented, but the feature alone does not show that it changes apparent mass or creates another band.
UniProt lists N-acetylmethionine at position 1 in its sequence numbering. This modification is documented, but the feature alone does not show that it changes apparent mass or creates another band.
Does this guide establish induction of UBE2S?
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 UBE2S Western blot?
Transfer · UBE2S is predicted to be 23.8 kDa. Use transfer conditions that retain proteins around 24 kDa on the membrane, and check transfer efficiency at that size. The supplied features do not identify a single optimal transfer protocol.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03045-2 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 UBE2S be quantified across samples?
Quantitation · Quantify the same validated band near 24 kDa across samples. UBE2S is listed as an APC/C component and associated with the cell cycle, but these features do not establish how its abundance changes under any particular condition.
Why does UBE2S appear near 24 kDa?
Interpretation · UniProt predicts 23.8 kDa, consistent with the observed band near 24 kDa. This agreement does not establish the band’s identity; confirm it with an appropriate specificity control.
The record lists one isoform, phosphoserine 173, N-acetylmethionine 1, and APC/C complex membership. None alone identifies an unexpected band or proves a visible shift. Check band specificity before assigning extra bands to UBE2S, a modification, or the complex.
The record lists one isoform, phosphoserine 173, N-acetylmethionine 1, and APC/C complex membership. None alone identifies an unexpected band or proves a visible shift. Check band specificity before assigning extra bands to UBE2S, a modification, or the complex.
Boster reagents
UBE2S 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-UBE2S Antibody Picoband®
Cat # A03045-2
The catalog reports one anti-UBE2S antibody, A03045-2, with reported human and rat reactivity. Its Western blot image shows an approximately 24 kDa band in human cell lysates and rat testis lysate. The supplied evidence does not include orthogonal validation.
Which to pick: A03045-2 is the only listed option and has a Western blot image. Its caption documents 293T, HeLa, Jurkat, and MCF-7 cell lysates and rat testis lysate, using 0.5 μg/mL primary antibody. Choose it for these documented contexts; other samples are not shown.
Source: BosterBio UBE2S gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16763.
- Human Protein Atlas. UBE2S tissue expression.
- PMC10523983 — target-verified WB comparison
- PMC6330942 — target-verified WB comparison
- PMC9969189 — target-verified WB comparison