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USP11 / Ubiquitin carboxyl-terminal hydrolase 11 · Western blot design guide
Design a Western Blot for USP11
Source-linked USP11 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-USP11 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
USP11 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 | ~109.8 kDa | |
| Observed band | ~110 kDa | |
| Gel | 5–20% (catalog M05225) | |
| Positive control | Cerebellum (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 | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked USP11 Western Blot Protocol Options
The M05225 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human SH-SY5Y, human Jurkat (catalog M05225) |
| Gel % | 5–20% (catalog M05225) |
| Load | 30 ug; reducing conditions (catalog M05225) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M05225) |
| Membrane | nitrocellulose membrane (catalog M05225) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M05225) |
| Primary antibody | M05225 · 1:500 (catalog M05225) |
| Primary incubation | overnight at 4°C (catalog M05225) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M05225) |
| Secondary incubation | 1.5 hour at RT (catalog M05225) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M05225) |
| Detection | ECL (catalog M05225) |
Section 2
What Is the Expected USP11 Western Blot Band Size?
USP11 is predicted at 109.8 kDa and observed at ~110 kDa; no distinct cause for the small difference is established.
| Band at ~110 kDa in whole-cell lysate | Matches the observed USP11 band and its 109.8 kDa predicted mass |
| Single dominant band near ~110 kDa | Consistent with monomeric USP11 |
| ~110 kDa band enriched in nuclear fraction | Consistent with predominantly nuclear USP11 |
| ~110 kDa band in cytoplasmic fraction | Consistent with the reported cytoplasmic location of USP11 |
💡Expected USP11 appearanceUSP11 has a predicted mass of 109.8 kDa and an observed band at ~110 kDa; confirm band identity with antibody specificity controls.
| Predicted USP11 mass | Sets a 109.8 kDa reference, close to the observed ~110 kDa band |
| N6-acetyllysine at K245 | Adds a modification, with no demonstrated visible migration change |
| Phosphoserine at S648 | Adds a modification, with no demonstrated visible migration change |
| Phosphoserine at S733 | Adds a modification, with no demonstrated visible migration change |
| Phosphoserine at S948 | Adds a modification, with no demonstrated visible migration change |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Predominantly nuclear USP11 may be dilute in whole-cell lysate | Check a nuclear fraction and a positive-control lysate |
| Band higher than expected | USP11 phosphorylation is documented, but its effect on migration is unestablished | Compare phosphatase-treated and untreated samples and verify band identity |
| Band lower than expected | A smaller band is not explained by the supplied USP11 processing features | Check sample integrity and confirm identity with an independent antibody |
| Multiple bands | Phosphorylated forms or nonspecific antibody binding are possible; distinct forms are unverified | Compare phosphatase treatment and an independent antibody |
| Fragments below expected size | Sample degradation could produce fragments below full-length USP11 | Prepare fresh lysate with protease inhibitors and verify antibody specificity |
Experiment evidence
Western blot images showing USP11 band
M05225 · USP11
M05225-1 · USP11
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for USP11 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for USP11 in Western blot, you can use cerebellum lysate.
Positive control: Cerebellum (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: USP11 is predominantly nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for USP11
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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - nucleus | High | Protein (IHC) | HPA → |
| Testis | spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced USP11 Western Blot Tips
Deeper troubleshooting and optimisation questions for USP11, answered from its protein features.
How should USP11 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 USP11 isoforms explain multiple bands?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. It provides no isoform-based explanation for multiple bands.
Which USP11 modifications should I consider when interpreting bands?
PTM · UniProt lists N6-acetyllysine at position 245 and phosphoserine at positions 648, 733, and 948. These are UniProt coordinates; paper or antibody numbering may differ. The listed sites do not establish that a separate band will appear.
Does this guide establish induction of USP11?
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 USP11 Western blot?
Transfer · USP11 is predicted to be 109.8 kDa. Choose transfer conditions suitable for a protein near 110 kDa and check that protein in this size range transfers effectively; the supplied features do not specify a membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M05225 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 USP11 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Does the observed USP11 band match its predicted mass?
Interpretation · The observed band near 110 kDa is close to the predicted 109.8 kDa. USP11 modifications are listed, but their presence alone does not establish a visible shift or explain any mass difference.
USP11 is reported in the nucleus, cytoplasm, and chromosomes, with predominantly nuclear localization noted. When quantifying fractions, compare like fractions across samples so a change in distribution is not mistaken for a change in total abundance.
Use the approximately 110 kDa band as a reference point. One isoform is listed, and no signal peptide or propeptide is supplied. The listed modifications do not by themselves identify an unexpected band; check whether the band is reproducible before assigning it to USP11.
USP11 is reported in the nucleus, cytoplasm, and chromosomes, with predominantly nuclear localization noted. When quantifying fractions, compare like fractions across samples so a change in distribution is not mistaken for a change in total abundance.
Use the approximately 110 kDa band as a reference point. One isoform is listed, and no signal peptide or propeptide is supplied. The listed modifications do not by themselves identify an unexpected band; check whether the band is reproducible before assigning it to USP11.
Boster reagents
USP11 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-USP11 Monoclonal Antibody
Cat # M05225
Real WB data
Anti-USP11 Antibody (C-term R565)
Cat # M05225-1
Two anti-USP11 antibodies have Western blot images. M05225 shows an approximately 110 kDa band in human cell, rat tissue, and mouse tissue lysates. M05225-1 shows blots of three human cell lines. The supplied evidence does not establish performance beyond these tested contexts.
Which to pick: For human, mouse, or rat samples, consider M05225, whose blot includes examples from all three species at 30 µg per lane and 1:500. For human cell lines, M05225-1 also has a blot using 35 µg per lane and 1:1000.
Source: BosterBio USP11 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P51784.
- Human Protein Atlas. USP11 tissue expression.
- PMC13014669 — target-verified WB comparison
- PMC8546420 — target-verified WB comparison
- PMC5749825 — target-verified WB comparison