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- Table of Contents
UCHL5 / Ubiquitin carboxyl-terminal hydrolase isozyme L5 · Western blot design guide
Design a Western Blot for UCHL5
Source-linked UCHL5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UCHL5 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
UCHL5 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 | ~37.6 kDa | |
| Gel | 12–15% (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 | Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked UCHL5 Western Blot Protocol Options
The A04276 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | various lysates, (catalog A04276) |
| Gel % | 12–15% (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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04276) |
| Primary antibody | A04276 · 1:433 (catalog A04276) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04276) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04276) |
Section 2
What Is the Expected UCHL5 Western Blot Band Size?
UCHL5 is predicted at 37.6 kDa; four isoforms and three lysine modifications are documented, but their effects on observed migration are unproven.
| Band near 37.6 kDa | consistent with the predicted UCHL5 mass; confirm identity with antibody controls |
| Several bands at different positions | could reflect isoforms 1, 2, 3, and 4; their migration differences are unknown |
| Band away from 37.6 kDa | could reflect an isoform, but band identity needs confirmation |
| Weak band in a cell fraction | UCHL5 occurs in both cytoplasm and nucleus, so recovery may depend on the fraction |
💡Expected UCHL5 appearanceUCHL5 has a predicted mass of 37.6 kDa, but no empirical band size is supplied; isoforms may alter migration, so confirm a candidate band with antibody controls.
| Predicted UCHL5 mass | 37.6 kDa is the sequence-based reference, not a measured band |
| Isoforms 1, 2, 3, and 4 | may differ in size; their masses and resolvable migration differences are unknown |
| N6-succinyllysine at residue 47 | no visible band shift is established |
| N6-acetyllysine at residue 158 | no visible band shift is established |
| N6-succinyllysine at residue 289 | no visible band shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | UCHL5 may be below detection in the sampled material | check a positive lysate and examine cytoplasmic and nuclear fractions |
| Band higher than expected | an isoform or an unrelated antibody target may migrate there | confirm identity by UCHL5 depletion and an independent antibody |
| Band lower than expected | an isoform or an unrelated antibody target may migrate there | confirm identity by UCHL5 depletion and an independent antibody |
| Multiple bands | four named isoforms are documented, but their band positions are unknown | compare bands after UCHL5 depletion |
| Weak or no signal | UCHL5 occurs in cytoplasm and nucleus, affecting recovery from either fraction | check both fractions and a positive lysate |
Experiment evidence
Western blot images showing UCHL5 band
A04276 · UCHL5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UCHL5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UCHL5 in Western blot, you can use adrenal gland lysate, which has high HPA expression.
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 is reported as not detected, so use siRNA knockdown or a KO line for a clear negative control.
HPA tissue expression evidence for UCHL5
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 → |
| Lymph node | germinal center cells | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Colon | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced UCHL5 Western Blot Tips
Deeper troubleshooting and optimisation questions for UCHL5, answered from its protein features.
Where should the main UCHL5 band appear?
Band shift · The predicted mass is 37.6 kDa. No observed Western blot band position is supplied, so use 37.6 kDa as a reference, not a guaranteed apparent mass.
Could UCHL5 isoforms produce different bands?
Isoforms · Four isoforms are listed. Isoforms 2, 3, and 4 lack canonical residue 246; isoform 2 also replaces residues 316–329 with GK, while isoform 4 replaces them with FEKHFEKTLLGK. These differences may affect migration, but the features do not establish visible bands.
Check whether its stated recognition region includes canonical residue 246 or residues 316–329. Those regions differ in the listed isoforms, so recognition may vary. The supplied features do not specify any antibody epitope.
Check whether its stated recognition region includes canonical residue 246 or residues 316–329. Those regions differ in the listed isoforms, so recognition may vary. The supplied features do not specify any antibody epitope.
Which UCHL5 modifications should I consider?
PTM · UniProt lists N6-succinyllysine at residues 47 and 289 and N6-acetyllysine at residue 158. These are UniProt sequence coordinates; antibody or paper numbering may differ. Their presence alone does not establish a visible band shift.
Does this guide establish induction of UCHL5?
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 UCHL5?
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 A04276 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 UCHL5 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.
How should I compare UCHL5 across cell fractions?
Interpretation · UCHL5 is reported in cytoplasm and nucleus: it associates with the proteasome 19S subunit in cytoplasm and the INO80 complex in nucleus. Quantify the same fraction across samples and identify which fraction was analyzed when reporting abundance.
Compare their positions with the 37.6 kDa predicted mass and consider the listed isoform sequence differences and modified residues. Migration alone cannot assign an unexpected band to an isoform or modification; no empirical band positions are supplied.
Compare their positions with the 37.6 kDa predicted mass and consider the listed isoform sequence differences and modified residues. Migration alone cannot assign an unexpected band to an isoform or modification; no empirical band positions are supplied.
Boster reagents
UCHL5 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-UCHL5 Antibody
Cat # A04276
A04276 is an anti-UCHL5 antibody listed for human, mouse, and rat. Its Western blot image uses various lysates at 25 µg per lane and a 1:433 primary dilution. The caption does not identify the lysates or their species.
Which to pick: A04276 is the only listed option and has a Western blot image. Check its listed reactivity against your sample species; the image caption does not specify which species were tested.
Source: BosterBio UCHL5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y5K5.
- Human Protein Atlas. UCHL5 tissue expression.
- PMC7300206 — target-verified WB comparison
- PMC11660788 — target-verified WB comparison
- PMC11110341 — target-verified WB comparison