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UBE2L3 / Ubiquitin-conjugating enzyme E2 L3 · Western blot design guide
Design a Western Blot for UBE2L3
Source-linked UBE2L3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBE2L3 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
UBE2L3 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 | ~17.9 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Testis (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 | Acetylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked UBE2L3 Western Blot Protocol Options
The M01832 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 M01832) |
| 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 | M01832; use the WB datasheet starting dilution (standard starting point) |
| 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 UBE2L3 Western Blot Band Size?
UBE2L3 is predicted at 17.9 kDa; isoforms could affect migration, but no empirical band size or distinct isoform pattern is supplied.
| Band near 17.9 kDa | consistent with the predicted UBE2L3 mass; confirm identity with a specific control |
| Several bands at different positions | could reflect isoforms 1, 2, and 3; their migration is not established |
| One band despite three annotated isoforms | the isoforms may not produce resolvable bands |
| Bands in nuclear and cytoplasmic fractions | consistent with UBE2L3 localization in both compartments |
💡Expected UBE2L3 appearanceUBE2L3 has a predicted mass of 17.9 kDa, but no empirical band size is supplied; confirm any candidate band with a specific identity control.
| UniProt predicted mass | places the reference size near 17.9 kDa |
| Splice isoform 1 | has no supplied isoform-specific mass or migration |
| Splice isoform 2 | may differ in size, but its mass and migration are unspecified |
| Splice isoform 3 | may differ in size, but its mass and migration are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| Multiple bands | annotated isoforms or unrelated antibody binding | compare with UBE2L3 depletion and check which bands change |
| Band higher than expected | an isoform or another detected protein; the cause is unestablished | verify identity with UBE2L3 depletion |
| Band lower than expected | an isoform or sample proteolysis; the cause is unestablished | compare fresh lysate with UBE2L3-depleted lysate |
| Weak or no signal | UBE2L3 may be distributed between nucleus and cytoplasm | check both fractions and confirm extraction and loading |
| Fragments below expected size | possible proteolysis during sample preparation | prepare fresh lysate with protease inhibitors and verify fragment identity |
Experiment evidence
Western blot images showing UBE2L3 band
M01832 · UBE2L3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBE2L3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBE2L3 in Western blot, you can use testis lysate, which has high HPA expression.
Positive control: Testis (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: Ovary is reported as not detected, making tissue controls feasible, though a faint signal may still require validation.
HPA tissue expression evidence for UBE2L3
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 |
|---|---|---|---|---|
| Testis | round or early spermatids | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | 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 → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Breast | adipocytes | Low | Protein (IHC) | HPA → |
Section 3
Advanced UBE2L3 Western Blot Tips
Deeper troubleshooting and optimisation questions for UBE2L3, answered from its protein features.
How should UBE2L3 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.
How could UBE2L3 isoforms affect band patterns?
Isoforms · UniProt lists three isoforms. Isoform 2 lacks canonical residues 10–41, while isoform 3 replaces canonical residues 1–9 with a longer sequence. These differences could affect migration, but the supplied features do not establish where either isoform runs.
Check whether the antibody recognizes a region shared by the isoforms before treating one band as total UBE2L3. Isoform 2 lacks canonical residues 10–41, and isoform 3 has a different N-terminal sequence; an N-terminal antibody may detect them differently.
Check whether the antibody recognizes a region shared by the isoforms before treating one band as total UBE2L3. Isoform 2 lacks canonical residues 10–41, and isoform 3 has a different N-terminal sequence; an N-terminal antibody may detect them differently.
Where is UBE2L3 acetylated?
PTM · UniProt lists N6-acetyllysine at position 131 in its canonical numbering. Check an antibody’s numbering convention before comparing sites. This modification alone does not establish a visible band shift.
Does HPV-16 E6 imply UBE2L3 induction?
Induction · No induction is listed. The supplied interaction feature says UBE2L3 interacts with UBE3A, which HPV-16 E6 uses to ubiquitinate p53/TP53. That pathway connection does not establish increased UBE2L3 abundance or a new UBE2L3 band.
How should transfer be checked for UBE2L3?
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 M01832 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.
Should nuclear and cytoplasmic UBE2L3 be quantified together?
Quantitation · UniProt places UBE2L3 in both nucleus and cytoplasm. Specify whether the measurement uses whole-cell lysate or a cellular fraction, and compare like fractions across samples.
Should UBE2L3 run at exactly 17.9 kDa?
Interpretation · 17.9 kDa is the predicted mass of the 154-residue canonical sequence. No observed band position is supplied, so the features cannot establish an apparent mass or explain a difference from 17.9 kDa.
Compare bands with the 17.9 kDa canonical prediction and the sequence changes listed for isoforms 2 and 3. An extra band cannot be assigned to an isoform or to Lys131 acetylation from these features alone.
Compare bands with the 17.9 kDa canonical prediction and the sequence changes listed for isoforms 2 and 3. An extra band cannot be assigned to an isoform or to Lys131 acetylation from these features alone.
Boster reagents
UBE2L3 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-Ube2L3 / UBCH7 Rabbit Monoclonal Antibody
Cat # M01832
M01832 is a rabbit monoclonal anti-UBE2L3/UBCH7 antibody listed for Western blotting, with reported human, mouse, and rat reactivity. Its supplied WB image shows Jurkat cell lysate; no other tested samples or validation evidence are provided.
Which to pick: M01832 is the only listed option and has a WB image from Jurkat cell lysate. Consider it for human samples matching that context; mouse and rat reactivity is listed, but the supplied WB example does not demonstrate those species.
Source: BosterBio UBE2L3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P68036.
- Human Protein Atlas. UBE2L3 tissue expression.
- PMC11043109 — target-verified WB comparison
- PMC5300903 — target-verified WB comparison