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- Table of Contents
USP10 / Ubiquitin carboxyl-terminal hydrolase 10 · Western blot design guide
Design a Western Blot for USP10
Real validated USP10 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-USP10 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
USP10 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 | ~87.1 kDa | |
| Gel | 8–10% (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 | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated USP10 Western Blot Protocols
The A03786 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 A03786) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03786 · (A) 1 and (B) 2 μg/mL (catalog A03786) |
| 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 USP10 Western Blot Band Size?
USP10 is predicted at 87.1 kDa; isoforms and phosphorylation could affect its pattern, but no empirical migration or distinct isoform bands are established.
| Band near 87.1 kDa | consistent with predicted USP10 size; confirm identity with controls |
| Several bands near the expected region | could reflect USP10 isoforms 1, 2, and 3; distinct migration is unproven |
| Close doublet | could reflect different USP10 phosphorylation states; migration effects are unproven |
| Stronger nuclear band after DNA damage | consistent with reported USP10 translocation to the nucleus |
💡Expected USP10 appearanceUniProt predicts USP10 at 87.1 kDa; no empirical band size is supplied, and isoform or phosphorylation effects on migration require band-identity controls.
| Predicted USP10 mass | sets an 87.1 kDa reference for the protein |
| USP10 isoform 1 | its individual mass and migration are not supplied |
| USP10 isoform 2 | alternative splicing could change size; relative mass is not supplied |
| USP10 isoform 3 | alternative splicing could change size; relative mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | USP10 signal may be below detection | check antibody performance with a positive lysate and load more protein |
| Band higher than expected | phosphorylation may affect migration, but the size effect is unestablished | confirm identity with an independent USP10 antibody or USP10 depletion |
| Band lower than expected | an alternative splice isoform could migrate differently; isoform sizes are unknown | confirm identity by USP10 depletion and an isoform-aware assay |
| Multiple bands | USP10 has three named isoforms and documented phosphorylation sites | compare bands after USP10 depletion; assess phosphorylation if needed |
| Weak or no signal | USP10 occupies cytoplasmic, nuclear, and early endosomal compartments and translocates after DNA damage | check the relevant fraction and use a positive lysate control |
Experiment evidence
Western blot images showing USP10 band
A03786 · USP10
M03786 · USP10
A03786-3 · USP10
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for USP10 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for USP10 in Western blot, you can use adrenal gland lysate, which HPA scores High.
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 listed is undetected, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for USP10
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 → |
| Appendix | 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Soft tissue | chondrocytes | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced USP10 Western Blot Tips
Deeper troubleshooting and optimisation questions for USP10, answered from its protein features.
Where should the main USP10 band appear?
Band shift · The predicted mass of canonical USP10 is 87.1 kDa. No observed band position is supplied, so use 87.1 kDa as a reference, not a guaranteed apparent mass. The listed modifications alone do not establish a visible shift or explain any difference from the prediction.
Could USP10 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to canonical isoform 1, isoform 2 replaces the initial methionine with a longer N-terminal sequence; isoform 3 replaces residues 1–6 (MALHSP) with MPWLPSPGIG. Check whether the antibody recognizes the relevant N-terminal sequence. These variants may differ in size, but the features do not establish their band positions.
Which USP10 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylalanine at position 2; phosphothreonine at 24, 42, and 100; and phosphoserine at 211, 226, 321, 337, 365, 370, 547, 563, and 576. ATM is specified for phosphorylation at 42 and 337. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not demonstrate a visible band shift.
Does this guide establish induction of USP10?
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 USP10?
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 A03786 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 USP10 be quantified across cell fractions?
Quantitation · USP10 is listed in the cytoplasm, nucleus, and early endosome. Quantify matched fractions consistently and account for fraction recovery before interpreting a change in one compartment as a change in total protein. Its reported movement to the nucleus after DNA damage makes compartment-specific measurements particularly relevant.
How should DNA damage affect the USP10 blot design?
Interpretation · USP10 is cytoplasmic under normal conditions, and the supplied location note reports nuclear translocation after DNA damage. Compare matched cytoplasmic and nuclear fractions if testing that response. ATM is listed for phosphorylation at UniProt positions 42 and 337. These features support assessing location and phosphorylation, but do not establish an increase in total USP10 abundance.
Compare them with the 87.1 kDa canonical prediction and consider the documented N-terminal changes in isoforms 2 and 3. UniProt lists phosphorylation and N-terminal acetylation, but these features alone cannot assign an extra band or prove a mobility shift. No glycosylation sites, signal peptide, or propeptide are supplied to support those explanations.
Compare them with the 87.1 kDa canonical prediction and consider the documented N-terminal changes in isoforms 2 and 3. UniProt lists phosphorylation and N-terminal acetylation, but these features alone cannot assign an extra band or prove a mobility shift. No glycosylation sites, signal peptide, or propeptide are supplied to support those explanations.
Boster reagents
USP10 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-USP10 Antibody
Cat # A03786
Real WB data
Anti-USP10 Monoclonal Antibody
Cat # M03786
Real WB data
Anti-USP10 Antibody Picoband®
Cat # A03786-3
All three listed anti-USP10 antibodies have WB images: A03786 in Jurkat lysate, M03786 in A375 lysate, and A03786-3 in the specified human cell and rat/mouse samples. A03786-3 shows bands near 110 and 130 kDa, versus an expected 87 kDa; band identity needs confirmation.
Which to pick: Choose by sample and stated reactivity: A03786-3 lists human, mouse, and rat and shows WB lanes for each; A03786 lists human and mouse but shows Jurkat lysate; M03786 lists human and shows A375 lysate. The images document those samples, not every listed species.
Source: BosterBio USP10 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14694.
- Human Protein Atlas. USP10 tissue expression.
- PMC12256264 — target-verified WB comparison
- PMC10033651 — target-verified WB comparison
- PMC11965059 — target-verified WB comparison