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USP13 / Ubiquitin carboxyl-terminal hydrolase 13 · Western blot design guide
Design a Western Blot for USP13
Source-linked USP13 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-USP13 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
USP13 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 | ~97.3 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked USP13 Western Blot Protocol Options
The M07816 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M07816) |
| 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 | M07816; 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 USP13 Western Blot Band Size?
USP13 has a predicted mass of 97.3 kDa; isoforms and phosphorylation are annotated, but their effects on band migration are not demonstrated.
| Band near 97.3 kDa | Consistent with the predicted size of USP13; confirm identity with controls. |
| Band at another size | Could represent a USP13 splice isoform; its migration is not supplied. |
| Several discrete bands | Isoforms 1 and 2 are annotated, but separate bands are not established. |
| Closely spaced doublet | Phosphorylation at Ser114 or Thr122 could contribute, but a migration shift is not established. |
💡Expected USP13 appearanceUniProt predicts 97.3 kDa for USP13; no empirical band size is supplied, and isoform or phosphorylation effects on migration require band-identity controls.
| UniProt predicted mass | Sets a 97.3 kDa reference for band assessment. |
| Alternative splicing | Produces named isoforms 1 and 2; their relative sizes are not supplied. |
| Isoform 1 | May migrate differently from isoform 2; its size is not supplied. |
| Isoform 2 | May migrate differently from isoform 1; its size is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | An isoform or modified USP13 is possible, but the cause is unproven. | Confirm identity with USP13 depletion and an independent antibody. |
| Band lower than expected | An isoform is possible; its size is not supplied. | Confirm identity with USP13 depletion and an independent antibody. |
| Multiple bands | USP13 has isoforms 1 and 2, though separate migration is unproven. | Compare isoform-specific expression samples or depletion controls. |
| Weak or no signal | USP13 abundance or antibody sensitivity may be low in the tested lysate. | Check a positive-control lysate, loading, and antibody performance. |
| Fragments below expected size | Sample proteolysis is possible; no cleavage product is annotated. | Prepare fresh lysate with protease inhibitors and confirm band identity. |
Experiment evidence
Western blot images showing USP13 band
M07816 · USP13
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for USP13 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for USP13 in Western blot, you can use adrenal gland lysate, based on its high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Cytoplasmic localization supports tissue lysate testing, and HPA identifies not-detected tissues for negative controls.
HPA tissue expression evidence for USP13
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 → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Testis | sertoli cells | High | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | High | 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 → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced USP13 Western Blot Tips
Deeper troubleshooting and optimisation questions for USP13, answered from its protein features.
How should USP13 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 USP13 isoforms produce different bands?
Isoforms · Yes. Isoform 2 lacks residues 1–65 of canonical isoform 1, so it has a shorter sequence. If both isoforms are expressed and detected, they may appear at different positions. The supplied features do not establish which isoform is present in a sample.
Which USP13 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphoserine at position 114, modified by AURKB, and phosphothreonine at position 122. These are canonical sequence coordinates; antibody or paper numbering may differ. Their annotation does not establish that either modification causes a visible band shift.
Does this guide establish induction of USP13?
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 USP13 Western blot?
Transfer · USP13 has a predicted mass of 97.3 kDa. Choose and verify transfer conditions suitable for a protein of approximately that size, checking that USP13 reaches the membrane. The supplied features do not identify a specific transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M07816 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 USP13 bands be quantified?
Quantitation · Quantify a consistently identified USP13 band across samples. Because isoform 2 lacks canonical residues 1–65, establish whether the antibody detects one or both isoforms before combining band intensities. Keep sample fraction and band selection consistent; USP13 is annotated in the cytoplasm.
Why might USP13 migrate differently from its predicted mass?
Interpretation · The supplied predicted mass is 97.3 kDa, but no observed band position is available. USP13 has two isoforms and two annotated phosphorylation sites; these features alone do not establish a visible shift or explain a mass difference.
Compare them with the 97.3 kDa prediction and consider the shorter isoform 2, which lacks canonical residues 1–65. Phosphorylation at canonical positions 114 and 122 is annotated, but cannot by itself identify an unexpected band. Confirm band identity before assigning an isoform or modification.
Compare them with the 97.3 kDa prediction and consider the shorter isoform 2, which lacks canonical residues 1–65. Phosphorylation at canonical positions 114 and 122 is annotated, but cannot by itself identify an unexpected band. Confirm band identity before assigning an isoform or modification.
Boster reagents
USP13 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-USP13 Monoclonal Antibody
Cat # M07816
M07816 is a monoclonal anti-USP13 antibody listed for Western blotting with Human and Mouse reactivity. Its supplied WB image shows USP13 expression in HepG2 cell lysate; no other validation contexts are provided.
Which to pick: M07816 is the only listed option. Its WB image uses HepG2 cell lysate, so it provides a documented example for that sample; Mouse reactivity is listed but no Mouse WB example is supplied.
Source: BosterBio USP13 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92995.
- Human Protein Atlas. USP13 tissue expression.
- PMC9818633 — target-verified WB comparison
- PMC7948425 — target-verified WB comparison
- PMC12639959 — target-verified WB comparison