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- Table of Contents
UFD1 / Ubiquitin recognition factor in ER-associated degradation protein 1 · Western blot design guide
Design a Western Blot for UFD1
Source-linked UFD1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-UFD1 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
UFD1 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 | ~34.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Breast (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 | 3 isoform(s) |
Section 1
Source-Linked UFD1 Western Blot Protocol Options
The M32358-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M32358-1) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M32358-1; 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 UFD1 Western Blot Band Size?
UFD1 is predicted at 34.5 kDa; annotated isoforms and phosphoserines could affect migration, but no empirical band size or feature-dependent shift is established.
| Band near 34.5 kDa | Consistent with the predicted UFD1 mass; confirm identity with controls |
| Several discrete bands | Could reflect the annotated Short, Long, and 3 isoforms; their migration is not established |
| Band in a cytosolic fraction | Consistent with annotated cytosolic localization |
| Band in a nuclear fraction | Consistent with annotated nuclear localization |
💡Expected UFD1 appearanceUFD1 has a predicted mass of 34.5 kDa, but no empirical band size is supplied; use knockdown or another band-identity control to assess any detected band.
| UniProt predicted mass | Places the reference band near 34.5 kDa |
| Short isoform | May differ in size from Long; its mass and migration are not supplied |
| Long isoform | May differ in size from Short; its mass and migration are not supplied |
| Isoform 3 | May differ in size from the other isoforms; its relative size is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear and cytosolic UFD1 may be poorly recovered by the extraction method | Check extraction controls and compare nuclear and cytosolic fractions |
| Band higher than expected | UFD1 has annotated phosphoserines, but an effect on migration is unproven | Compare matched phosphatase-treated samples and confirm band identity |
| Band lower than expected | An alternative isoform is possible, but its mass is not supplied | Confirm the band with UFD1 knockdown or an independent antibody |
| Multiple bands | Short, Long, and 3 isoforms are annotated, but distinct bands are unproven | Use UFD1 knockdown to identify specific bands |
| Weak or no signal | Nuclear and cytosolic distribution may reduce signal in an individual fraction | Compare whole-cell lysate with controlled nuclear and cytosolic fractions |
Experiment evidence
Western blot images showing UFD1 band
M32358-1 · UFD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for UFD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for UFD1 in Western blot, you can use breast tissue, which HPA rates High.
Positive control: Breast (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: HPA lists a not-detected tissue, but confirm its lack of signal in your lysate.
HPA tissue expression evidence for UFD1
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 |
|---|---|---|---|---|
| Breast | myoepithelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | High | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (ciliary rootlets) | Medium | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UFD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for UFD1, answered from its protein features.
How should UFD1 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.
Which UFD1 isoforms could produce different band sizes?
Isoforms · Three isoforms are listed: Short, Long, and 3. Long replaces E at UniProt position 106 with a longer sequence; isoform 3 lacks UniProt residues 267–307. These sequence differences could affect band size, but no isoform-specific observed masses are supplied.
Isoform 3 lacks UniProt residues 267–307, which include the annotated phosphoserine at position 299. A signal assigned to that site should therefore be interpreted in light of which isoform the assay detects.
Isoform 3 lacks UniProt residues 267–307, which include the annotated phosphoserine at position 299. A signal assigned to that site should therefore be interpreted in light of which isoform the assay detects.
Which UFD1 phosphorylation sites are annotated?
PTM · Phosphoserine is annotated at UniProt positions 129, 231, 245, 247, and 299. Match any phospho-specific antibody’s stated numbering convention to these UniProt coordinates before interpreting its signal.
No. UFD1 has N-acetylmethionine at UniProt position 1 and five annotated phosphoserines, but these features alone do not demonstrate a visible shift or explain a difference from 34.5 kDa.
No. UFD1 has N-acetylmethionine at UniProt position 1 and five annotated phosphoserines, but these features alone do not demonstrate a visible shift or explain a difference from 34.5 kDa.
Does this guide establish induction of UFD1?
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 UFD1?
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 M32358-1 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 UFD1 bands be quantified across samples?
Quantitation · Check whether the antibody recognizes all three isoforms, especially where Long changes position 106 and isoform 3 lacks residues 267–307. Compare corresponding bands consistently. If using nuclear and cytosolic fractions, quantify them separately; UFD1 is annotated in both locations.
How should a UFD1 band be compared with predicted mass?
Interpretation · The predicted mass is 34.5 kDa, but no observed band position is supplied. Compare the band with this reference while considering the annotated Long and isoform 3 sequence changes. Their presence does not establish a particular apparent mass.
Compare their positions with the 34.5 kDa predicted mass and the annotated Long and isoform 3 sequence changes. Check antibody recognition of the altered regions. The supplied features and absent observed-band reference cannot assign an unexpected band to an isoform or modification by themselves.
Compare their positions with the 34.5 kDa predicted mass and the annotated Long and isoform 3 sequence changes. Check antibody recognition of the altered regions. The supplied features and absent observed-band reference cannot assign an unexpected band to an isoform or modification by themselves.
Boster reagents
UFD1 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-UFD1L Rabbit Monoclonal Antibody
Cat # M32358-1
The listed anti-UFD1 antibody, M32358-1, is a rabbit monoclonal with reported Human, Mouse, and Rat reactivity. Its Western blot image shows UFD1L expression in HeLa cell lysate; no other specimen or condition is shown here.
Which to pick: M32358-1 is the only listed option for UFD1. It has a Western blot image from HeLa cell lysate. Check the reported reactivity against your sample species; the image documents a HeLa test, not validation across all three listed species.
Source: BosterBio UFD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92890.
- Human Protein Atlas. UFD1 tissue expression.
- PMC9100061 — target-verified WB comparison
- PMC9397562 — target-verified WB comparison
- PMC13542637 — target-verified WB comparison