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UBA52 / Ubiquitin-ribosomal protein eL40 fusion protein · Western blot design guide
Design a Western Blot for UBA52
Source-linked UBA52 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBA52 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
UBA52 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 | ~14.7 kDa | |
| Observed band | ~15 kDa | |
| Gel | 12% (catalog M04905-1) | |
| 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 + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked UBA52 Western Blot Protocol Options
The M04905-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human THP-1, human RT4, human MCF-7, rat small intestine, rat brain, mouse small intestine, mouse brain (catalog M04905-1) |
| Gel % | 12% (catalog M04905-1) |
| Load | 30 ug; reducing conditions (catalog M04905-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04905-1) |
| Membrane | nitrocellulose membrane (catalog M04905-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04905-1) |
| Primary antibody | M04905-1; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:500 (catalog M04905-1) |
| Secondary incubation | 1.5 hour at RT (catalog M04905-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04905-1) |
| Detection | ECL (catalog M04905-1) |
Section 2
What Is the Expected UBA52 Western Blot Band Size?
UBA52 is predicted at 14.7 kDa and observed at ~15 kDa; the cause of the small difference is not established by the supplied evidence.
| Band at ~15 kDa | matches the empirical UBA52 band in reducing whole-cell lysates |
| Band near 14.7 kDa | is consistent with the predicted mass of the 128-residue fusion protein |
| Band near ~15 kDa in cytoplasmic material | is consistent with the listed cytoplasmic location of ubiquitin |
| Band near ~15 kDa in nuclear material | is consistent with the listed nuclear location of ubiquitin |
💡Expected UBA52 appearanceUniProt predicts 14.7 kDa for the UBA52 fusion protein, and antibody QC reports a ~15 kDa band in reducing whole-cell lysates; confirm band identity with appropriate controls.
| Predicted UBA52 fusion-protein mass | sets a 14.7 kDa reference; the empirical band is ~15 kDa |
| Ser65 phosphorylation by PINK1 | adds a modification, with no demonstrated resolvable band shift |
| Thr66 infection-associated ADP-ribosylation | may modify mass during microbial infection, with no demonstrated band shift |
| Gly76 ADP-ribosylation | adds a modification, with no demonstrated resolvable band shift |
| Lys98 trimethylation | adds a modification, with no demonstrated resolvable band shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | UBA52 signal may be below detection or lost during sample preparation | check loading and transfer, then compare with a positive whole-cell lysate |
| Band higher than expected | the supplied features do not establish an additional higher-mass UBA52 band | compare with the ~15 kDa reference and verify identity with an independent antibody or UBA52 depletion |
| Band lower than expected | the supplied features do not identify a lower-mass product | check sample integrity and verify the band with UBA52 depletion |
| Multiple bands | additional bands cannot be assigned to distinct UBA52 isoforms from the supplied evidence | compare each band with the ~15 kDa reference and test specificity with UBA52 depletion |
| Weak or no signal | insufficient detectable UBA52 or inefficient transfer of a small protein | check lysate loading, transfer and a positive-control lysate |
| Fragments below expected size | sample breakdown is possible, but no fragment masses are supplied | prepare fresh lysate and verify any lower band with an independent antibody or UBA52 depletion |
Experiment evidence
Western blot images showing UBA52 band
M04905-1 · UBA52
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBA52 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBA52 in Western blot, you can use adrenal gland lysate.
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 negative control.
HPA tissue expression evidence for UBA52
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 → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Colon | endothelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced UBA52 Western Blot Tips
Deeper troubleshooting and optimisation questions for UBA52, answered from its protein features.
How should UBA52 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.
Do UBA52 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to UBA52 isoforms from this evidence.
Which UBA52 modification sites matter when interpreting bands?
PTM · The supplied UniProt coordinates are Ser65 (phosphorylated by PINK1), Thr66 (ADP-ribosylated during microbial infection), Gly76 (ADP-ribosylated), and Lys98 (trimethylated). These are coordinates in the supplied 128-residue sequence; antibody or paper numbering may differ. Their presence does not establish a visible band shift.
PINK1 is listed as the enzyme for phosphorylation at Ser65. That identifies a modification to investigate, but the feature alone cannot explain an observed mass difference or confirm the identity of a shifted band.
PINK1 is listed as the enzyme for phosphorylation at Ser65. That identifies a modification to investigate, but the feature alone cannot explain an observed mass difference or confirm the identity of a shifted band.
Does this guide establish induction of UBA52?
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 UBA52?
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 M04905-1 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 UBA52 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.
Is a ~15 kDa UBA52 band consistent with its predicted mass?
Interpretation · Yes. The predicted mass is 14.7 kDa, close to the reported ~15 kDa band. The listed modifications alone do not establish a visible mobility shift.
The Thr66 feature is specifically annotated for microbial infection. Consider that condition when investigating Thr66 modification; the annotation does not establish that a shifted band will appear.
Start with the ~15 kDa band expected from the 14.7 kDa fusion protein. The record also describes ubiquitin conjugation and eL40 membership in the 60S ribosomal subunit, but it does not identify any additional Western-blot band. Verify an unexpected band's identity before assigning a cause.
The Thr66 feature is specifically annotated for microbial infection. Consider that condition when investigating Thr66 modification; the annotation does not establish that a shifted band will appear.
Start with the ~15 kDa band expected from the 14.7 kDa fusion protein. The record also describes ubiquitin conjugation and eL40 membership in the 60S ribosomal subunit, but it does not identify any additional Western-blot band. Verify an unexpected band's identity before assigning a cause.
Boster reagents
UBA52 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-UBA52 Rabbit Monoclonal Antibody
Cat # M04905-1
M04905-1 is a rabbit monoclonal anti-UBA52 antibody with reported human, mouse, and rat reactivity. Its associated WB caption describes human cell and rodent tissue lysates, but names a different antibody SKU and reports a band near 15 kDa versus 10 kDa expected.
Which to pick: M04905-1 is the only listed option. It has an associated WB image, but the caption identifies a different antibody SKU. Confirm that the image applies to M04905-1 and assess the reported band size before choosing it.
Source: BosterBio UBA52 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P62987.
- Human Protein Atlas. UBA52 tissue expression.
- PMC12839166 — target-verified WB comparison
- PMC9738938 — target-verified WB comparison