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UBE2C / Ubiquitin-conjugating enzyme E2 C · Western blot design guide
Design a Western Blot for UBE2C
Real validated UBE2C Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBE2C 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
UBE2C 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 | ~19.7 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Adrenal gland (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 | 4 isoform(s) |
Section 1
Real Curated UBE2C Western Blot Protocols
The M01491-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | MCF7 cell lysate (catalog M01491-1) |
| 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 | M01491-1; 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 UBE2C Western Blot Band Size?
UBE2C is predicted at 19.7 kDa; alternative splicing and listed modifications may affect migration, but no empirical band or demonstrated shift is supplied.
| Band near 19.7 kDa | Consistent with the predicted UBE2C mass; confirm identity with a specific control. |
| Several bands at different positions | Isoforms 1, 2, 3 and 4 could contribute if expressed and resolved; their migration is not supplied. |
| Doublet near 19.7 kDa | Isoforms or Ser3 phosphorylation could contribute, but neither a visible split nor its cause is established. |
| Band away from 19.7 kDa | An isoform is possible, but its size and the band's identity require confirmation. |
💡Expected UBE2C appearanceUBE2C has a predicted mass of 19.7 kDa, but no empirical band size or isoform migration is supplied; confirm a candidate band with a UBE2C-specific control.
| Predicted UBE2C mass | 19.7 kDa is the sequence-based reference, not a measured band position. |
| Isoform 1 | Its individual mass and migration are not supplied. |
| Isoform 2 | Alternative splicing may change its size; its relative migration is not supplied. |
| Isoforms 3 and 4 | Alternative splicing may change their sizes; their relative migration is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | UBE2C signal may be below detection or the assay may have failed. | Check transfer and antibody performance with an MCF7 lysate positive control. |
| Band higher than expected | An isoform or unrelated antibody signal is possible; the cause is unestablished. | Confirm identity by UBE2C knockdown and compare with a positive control. |
| Band lower than expected | An alternative isoform or degradation is possible; no fragment size is supplied. | Use fresh lysate and confirm the band by UBE2C knockdown. |
| Multiple bands | Four named isoforms exist, but their resolution on a blot is unknown. | Check which bands decrease after UBE2C knockdown. |
| Weak or no signal | Target abundance or assay sensitivity may be insufficient. | Compare with MCF7 lysate and check loading, transfer and antibody conditions. |
Experiment evidence
Western blot images showing UBE2C band
M01491-1 · UBE2C
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBE2C Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBE2C in Western blot, you can use Appendix tissue, the top HPA positive candidate.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adrenal gland (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA rates Appendix expression as medium, so the positive signal may be modest.
HPA tissue expression evidence for UBE2C
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 | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBE2C Western Blot Tips
Deeper troubleshooting and optimisation questions for UBE2C, answered from its protein features.
How should UBE2C 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 UBE2C isoforms produce different bands?
Isoforms · Four isoforms are listed. Relative to canonical isoform 1, isoform 2 lacks residues 1–39, isoform 3 lacks 44–72, and isoform 4 replaces 73–140 with a different sequence. These differences could affect migration, but the features do not establish which isoforms appear as bands.
An epitope within canonical residues 1–39 is absent from isoform 2; one within 44–72 is absent from isoform 3. Isoform 4 replaces residues 73–140. Check the antibody's stated epitope against these canonical UniProt coordinates before treating a missing band as absence of UBE2C.
An epitope within canonical residues 1–39 is absent from isoform 2; one within 44–72 is absent from isoform 3. Isoform 4 replaces residues 73–140. Check the antibody's stated epitope against these canonical UniProt coordinates before treating a missing band as absence of UBE2C.
Which UBE2C modifications should guide band interpretation?
PTM · UniProt lists N-acetylalanine at position 2 and phosphoserine at position 3, using its canonical sequence coordinates. Neither feature alone establishes a resolvable band shift. Check the numbering convention before comparing these sites with antibody or paper annotations.
Does this guide establish induction of UBE2C?
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 UBE2C?
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 M01491-1 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 cell-cycle context affect UBE2C quantitation?
Quantitation · UBE2C is annotated with cell cycle, cell division, and mitosis keywords and as a component of the APC/C complex. Record and match cell-cycle context across samples when comparing band intensity; these annotations do not establish a specific induction condition or expression change.
Should UBE2C migrate exactly at its predicted 19.7 kDa?
Interpretation · The canonical sequence has a predicted mass of 19.7 kDa, but no observed band position is supplied. The listed modifications and isoforms alone do not establish a visible shift or explain any difference between apparent and predicted mass.
Compare bands with the four annotated isoform sequences and the antibody epitope. UBE2C also has listed modifications at canonical positions 2 and 3, but their presence does not identify an extra band. The supplied features do not establish the identity of any observed band.
Compare bands with the four annotated isoform sequences and the antibody epitope. UBE2C also has listed modifications at canonical positions 2 and 3, but their presence does not identify an extra band. The supplied features do not establish the identity of any observed band.
Boster reagents
UBE2C 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-UBE2C Rabbit Monoclonal Antibody
Cat # M01491-1
The catalog reports one anti-UBE2C antibody for Western blot: rabbit monoclonal M01491-1. It is listed as reactive with human samples and has a WB image showing UBE2C expression in MCF7 cell lysate. The supplied evidence covers that tested context only.
Which to pick: M01491-1 is the only listed option. Its human reactivity and MCF7 lysate WB image make it the relevant choice for a similar human cell lysate experiment; other sample contexts are not documented here.
Source: BosterBio UBE2C gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O00762.
- Human Protein Atlas. UBE2C tissue expression.
- PMC5602297 — target-verified WB comparison
- PMC8268282 — target-verified WB comparison
- PMC3974821 — target-verified WB comparison