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UBR1 / E3 ubiquitin-protein ligase UBR1 · Western blot design guide
Design a Western Blot for UBR1
Source-linked UBR1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UBR1 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
UBR1 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 | ~200.2 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Caudate (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked UBR1 Western Blot Protocol Options
The A03250-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | the lysates from HUVECcells (catalog A03250-1) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A03250-1; 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 UBR1 Western Blot Band Size?
UBR1 has a predicted full-length mass of 200.2 kDa; isoforms and phosphorylation are annotated, but their effects on band migration are not demonstrated.
| Single band near 200.2 kDa | consistent with the predicted full-length UBR1 mass; confirm identity with antibody controls |
| Band at a different mass | could represent isoform 1 or 2; their masses and migration are unspecified |
| Two bands at different masses | could reflect the two named isoforms, but distinct bands are not established |
| Close doublet near 200.2 kDa | phosphorylation at Thr21 or Ser1179 is documented, but an effect on migration is unproven |
💡Expected UBR1 appearanceFull-length UBR1 has a predicted mass of 200.2 kDa; no empirical band position or isoform migration is supplied, so confirm any candidate band with antibody specificity controls.
| Predicted full-length mass | sets a reference of 200.2 kDa, not a measured band position |
| Alternative splicing | produces isoforms 1 and 2, whose relative sizes are unspecified |
| Isoform 1 | may migrate differently from isoform 2; its mass is unspecified |
| Isoform 2 | may migrate differently from isoform 1; its mass is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | cytosolic UBR1 may be below detection or poorly recovered | check lysate quality, loading, and antibody performance with a positive control |
| Band higher than expected | the supplied features do not establish a higher migrating UBR1 species | check antibody specificity and compare with an independent UBR1 antibody or depletion control |
| Band lower than expected | an alternative isoform is possible, but its mass is unspecified | test band identity with UBR1 depletion or an independent antibody |
| Multiple bands | isoforms 1 and 2 are annotated, but distinct migration is unverified | compare bands after UBR1 depletion and check antibody specificity |
| Weak or no signal | low UBR1 abundance or inadequate detection may limit the cytosolic signal | check loading, transfer, antibody conditions, and a positive control |
Experiment evidence
Western blot images showing UBR1 band
A03250-1 · UBR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UBR1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UBR1 in Western blot, you can use caudate tissue, which HPA scores as High.
Positive control: Caudate (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: Adipose tissue is an HPA not-detected comparator, but confirm the absence of signal in your lysate.
HPA tissue expression evidence for UBR1
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Pancreas | exocrine glandular 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 → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for UBR1, answered from its protein features.
How should UBR1 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 UBR1 isoforms produce different bands?
Isoforms · Yes. UniProt lists isoforms 1 and 2. In isoform 2, canonical residues 795–803 change from NNETGLENV to TRCIRPWSL, and residues 804–1749 are missing. Isoform 2 should therefore be substantially smaller, but these features do not establish its apparent band position.
Determine whether the antibody recognizes a region shared by both isoforms or the canonical region missing from isoform 2 (residues 804–1749). Quantify distinct bands separately when possible, and report which isoform or region the measurement represents.
Determine whether the antibody recognizes a region shared by both isoforms or the canonical region missing from isoform 2 (residues 804–1749). Quantify distinct bands separately when possible, and report which isoform or region the measurement represents.
Which UBR1 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylalanine at position 2, phosphothreonine at 21, and phosphoserine at 1179, using canonical UniProt coordinates. Isoform 2 lacks the region containing position 1179. These annotations identify possible modification states; they do not establish that any produces a visible band shift.
Does this guide establish induction of UBR1?
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 UBR1 Western blot?
Transfer · Canonical UBR1 is predicted at 200.2 kDa, so assess transfer efficiency in the high-mass region when optimizing the method. The supplied features do not identify a specific membrane, transfer format, or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A03250-1 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 UBR1 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 a band differ from UBR1’s predicted mass be interpreted?
Interpretation · The canonical 1,749-residue protein has a predicted mass of 200.2 kDa. No observed band position is supplied. Check whether the band could represent isoform 2, which lacks residues 804–1749; the listed modifications alone do not establish a visible shift or explain a mass difference.
Consider isoform 2, which retains the first 794 canonical residues, substitutes residues 795–803, and lacks residues 804–1749. Check antibody epitope coverage before assigning a band. The supplied features give no observed band sizes and no glycosylation sites, so band identity needs experimental confirmation.
UBR1 is annotated in the cytoplasm and cytosol. Include a cytosolic fraction when comparing sample preparations, and interpret a weak signal in another fraction in light of that localization.
Consider isoform 2, which retains the first 794 canonical residues, substitutes residues 795–803, and lacks residues 804–1749. Check antibody epitope coverage before assigning a band. The supplied features give no observed band sizes and no glycosylation sites, so band identity needs experimental confirmation.
UBR1 is annotated in the cytoplasm and cytosol. Include a cytosolic fraction when comparing sample preparations, and interpret a weak signal in another fraction in light of that localization.
Boster reagents
UBR1 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-Ubr1 Antibody
Cat # A03250-1
The catalog reports A03250-1, an anti-UBR1 antibody with reported human and mouse reactivity. Its Western blot image shows HUVEC cell lysate; the supplied evidence does not show a mouse Western blot.
Which to pick: A03250-1 is the only listed option. It has a Western blot image using HUVEC cell lysate. For mouse samples, reactivity is listed, but no mouse Western blot example is supplied.
Source: BosterBio UBR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8IWV7.
- Human Protein Atlas. UBR1 tissue expression.
- PMC12312037 — target-verified WB comparison