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- Table of Contents
UBQLN2 / Ubiquilin-2 · Western blot design guide
Design a Western Blot for UBQLN2
Source-linked UBQLN2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBQLN2 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
UBQLN2 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 | ~65.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked UBQLN2 Western Blot Protocol Options
The A02508 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02508) |
| 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 A02508) |
| Primary antibody | A02508 · 1:1000 (catalog A02508) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A02508) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A02508) |
Section 2
What Is the Expected UBQLN2 Western Blot Band Size?
UBQLN2 is predicted at 65.7 kDa; homodimers and other complexes are possible, but their visible migration is not demonstrated by the supplied evidence.
| Band near 65.7 kDa | Consistent with the predicted UBQLN2 monomer; confirm band identity with controls |
| Band near 131 kDa | Could represent a UBQLN2 homodimer if the complex survives electrophoresis |
| Additional higher band | Could represent a retained complex with UBQLN1; its migration is not established |
| Weak band in a soluble fraction | UBQLN2 can also localize to the nucleus, membranes, and autophagosomes |
💡Expected UBQLN2 appearanceUBQLN2 has a predicted monomer mass of 65.7 kDa, but no empirical band size is supplied; confirm any candidate band with ordinary identity controls.
| Predicted monomer mass | Sets a 65.7 kDa reference, without establishing apparent migration |
| Homodimer formation | Could produce a band near twice the monomer mass if the complex persists during electrophoresis |
| Heterodimer formation with UBQLN1 | Could produce another higher band if the complex persists; its size is not supplied |
| N-acetylalanine at residue 2 | Is documented, but no detectable band shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | A retained UBQLN2 homodimer is possible | Compare reducing and denaturing conditions and confirm UBQLN2 identity |
| Multiple bands | Retained UBQLN2 homodimers or UBQLN1 complexes are possible | Compare denaturing conditions and verify each band with an independent UBQLN2 antibody |
| Weak or no signal | UBQLN2 may partition among cytoplasm, nucleus, membranes, and autophagosomes | Check extraction and fraction recovery with suitable controls |
| No band in lysate | The extraction method may recover UBQLN2 poorly from some compartments | Check lysate recovery and antibody performance with a positive control |
| Fragments below expected size | Sample degradation is possible; no physiological cleavage is annotated | Use fresh, protease-inhibited samples and confirm fragment identity |
Experiment evidence
Western blot images showing UBQLN2 band
A02508 · UBQLN2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBQLN2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBQLN2 in Western blot, you can use appendix lysate.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside samples.
⚠️Feasibility: No HPA tissue is listed as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for UBQLN2
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial 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 → |
| Salivary gland | glandular cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Testis | Leydig cells | Low | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced UBQLN2 Western Blot Tips
Deeper troubleshooting and optimisation questions for UBQLN2, answered from its protein features.
Where should the main UBQLN2 band appear?
Band shift · The predicted mass is 65.7 kDa. No observed band position is supplied, so use 65.7 kDa as a reference, not a guaranteed migration position.
Could an isoform explain a second band?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning a second band to a UBQLN2 isoform.
Does UBQLN2 acetylation cause a visible band shift?
PTM · The listed modification is N-acetylalanine at UniProt position 2. The feature alone does not establish a visible shift or explain a difference between apparent and predicted mass. Keep the UniProt coordinate convention when comparing antibody or paper numbering.
Does this guide establish induction of UBQLN2?
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 UBQLN2?
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 A02508 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 UBQLN2 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.
Could a higher band represent a UBQLN2 complex?
Interpretation · UBQLN2 forms homodimers and heterodimers with UBQLN1 and interacts with other proteins. These associations make a complex worth investigating, but the feature record cannot identify an unexpected band or establish whether a complex survives sample preparation.
UBQLN2 is listed in the cytoplasm, nucleus, membrane and autophagosomes. A signal in any of these fractions is compatible with its annotated locations. Interpret fraction signals alongside fraction purity rather than assigning a band to one location from size alone.
Because UBQLN2 has several annotated locations, compare signals within the same fraction using consistent loading and fraction-specific normalization. A change in one fraction could reflect redistribution; it does not by itself establish a change in total UBQLN2 abundance.
UBQLN2 is listed in the cytoplasm, nucleus, membrane and autophagosomes. A signal in any of these fractions is compatible with its annotated locations. Interpret fraction signals alongside fraction purity rather than assigning a band to one location from size alone.
Because UBQLN2 has several annotated locations, compare signals within the same fraction using consistent loading and fraction-specific normalization. A change in one fraction could reflect redistribution; it does not by itself establish a change in total UBQLN2 abundance.
Boster reagents
UBQLN2 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-Ubiquilin-2 UBQLN2 Antibody
Cat # A02508
The catalog reports A02508, an anti-UBQLN2 antibody with stated human and mouse reactivity and a Western blot image. Its caption reports cell-line extracts at 1:1000 and 25 µg per lane, but does not identify the cell lines.
Which to pick: A02508 is the only listed antibody. It has a WB image and stated human and mouse reactivity; compare its reported sample and blot conditions with your planned experiment.
Source: BosterBio UBQLN2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UHD9.
- Human Protein Atlas. UBQLN2 tissue expression.
- PMC9823102 — target-verified WB comparison
- PMC7873701 — target-verified WB comparison
- PMC12950104 — target-verified WB comparison