Customize ufd-1 Antibody

Description

Introduction to UFD1 Antibody

UFD1 antibodies are immunoreagents designed to detect and quantify the UFD1 protein, which forms a ternary complex with Valosin-Containing Protein (VCP) and Nuclear Protein Localization 4 (NPLOC4). This complex facilitates the retrotranslocation of misfolded proteins from the ER to the cytosol for proteasomal degradation . UFD1 is upregulated in aggressive cancers like T-cell acute lymphoblastic leukemia (T-ALL) and is implicated in MYC-driven oncogenesis .

Role in ER Stress and Apoptosis

ERAD Dysregulation: UFD1 inactivation exacerbates ER stress by impairing the clearance of misfolded proteins, leading to PERK-mediated apoptosis via upregulation of CHOP and downregulation of BCL2/BCLxL .
Cancer Relevance: In MYC-driven T-ALL, UFD1 heterozygosity reduces tumor burden by inducing caspase-3 activation and G0/G1 cell cycle arrest .

Table 1: Functional Impacts of UFD1 Modulation

Model System	Effect of UFD1 Loss	Key Outcomes
Zebrafish T-ALL	Increased apoptosis, reduced proliferation	Delayed tumor progression
Human T-ALL cells	PERK/CHOP activation, ER stress amplification	Tumor cell death
HeLa cells	Skp2 downregulation, p27 accumulation	Cell cycle delay

Applications in Research

UFD1 antibodies are widely used in:

Western Blot (WB): Detects endogenous UFD1 at ~35–40 kDa in human, mouse, and rat samples .
Immunoprecipitation (IP): Isolates UFD1-containing complexes for functional studies .
Immunofluorescence (IF): Localizes UFD1 to the ER and nucleus .

Table 2: Common UFD1 Antibody Applications

Application	Recommended Clones	Species Reactivity
Western Blot	EPR12847 (Abcam), #13789 (Cell Signaling)	Human, Mouse, Rat
Flow Cytometry	EPR12847 (Abcam)	Human
Immunohistochemistry	B-7, E-4 (Santa Cruz)	Human, Mouse

Future Directions

UFD1 antibodies are pivotal for exploring:

Therapeutic Targeting: Inhibiting UFD1 in MYC-driven cancers to trigger ER stress-mediated apoptosis .
Cell Cycle Mechanisms: Dissecting UFD1’s role in Skp2-p27 regulation and mitotic exit .

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Components: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 weeks (Made-to-order)

Synonyms

ufd-1 antibody; F19B6.2 antibody; Ubiquitin fusion degradation protein 1 homolog antibody; UB fusion protein 1 antibody

Target Names

ufd-1

Uniprot No.

Q19584

Target Background

Function

The ufd-1 antibody targets a protein functioning in a post-ubiquitination step within the ubiquitin fusion degradation (UFD) pathway. It interacts with npl-4.1 and/or npl-4.2, and the ATPases cdc-48.1 and/or cdc-48.2, facilitating the cytoplasmic degradation of misfolded proteins exported from the endoplasmic reticulum (ER). This process, known as ER-associated degradation (ERAD), prevents the unfolded protein response (UPR) activation caused by ER misfolded protein accumulation. During the S phase of the cell cycle, ufd-1, in conjunction with npl-4.1 and/or npl-4.2, cdc-48.1 and/or cdc-48.2, and ubxn-3, ensures the degradation of the DNA licensing factor cdt-1 following DNA replication initiation. This action contributes to the disassembly of the CMG helicase complex by promoting the chromatin dissociation of several components, including cdc-45 and sld-5. Furthermore, ufd-1 regulates ubxn-3 nuclear localization during S phase.

Database Links

KEGG: cel:CELE_F19B6.2

STRING: 6239.F19B6.2a

UniGene: Cel.22547

Protein Families

UFD1 family

Subcellular Location

Cytoplasm. Nucleus.

Q&A

Basic Research Questions

How should researchers validate UFD1 antibody specificity in Western blotting (WB) and immunoprecipitation (IP)?

Methodological Answer:
- Perform WB using lysates from UFD1-knockout cell lines to confirm absence of the ~40 kDa band.
- Use tissues/cells with known UFD1 expression levels (e.g., cancerous vs. normal T-cells) as positive/negative controls .
- For IP, pair with mass spectrometry to verify co-precipitated ERAD complex components (e.g., p97/VCP, NPL4) .

Validation Parameter	Recommended Approach
Specificity	Knockout validation + cross-reactivity testing (human, mouse, rat, monkey)
Sensitivity	Titrate antibody (1:500–1:2000 dilution) against endogenous UFD1 in HeLa/T-ALL cell lysates
Functional Confirmation	Correlate UFD1 depletion with ERAD impairment (e.g., accumulation of CHOP, GRP78)

What experimental models are optimal for studying UFD1’s role in ER-associated degradation (ERAD)?

Methodological Answer:
- Use MYC-overexpressing T-ALL cell lines to mimic pathological ER stress .
- Combine RNAi/CRISPR-mediated UFD1 knockdown with ERAD reporters (e.g., GFP-tagged misfolded proteins) .
- Monitor downstream markers: PERK phosphorylation, CHOP upregulation, and BCL2/BCLxL downregulation .

Advanced Research Questions

How can contradictory data on UFD1’s role in ERAD (proteasomal degradation acceleration vs. impairment) be resolved?

Methodological Answer:
- Conduct parallel experiments in yeast vs. mammalian systems to isolate species-specific effects .
- Compare RNAi (partial knockdown) vs. CRISPR (complete knockout) outcomes in the same cell line .
- Assess ERAD substrate specificity: Test cholera toxin (accelerated degradation) vs. MHC class I heavy chain (impaired degradation) .

Study System	ERAD Outcome	Proposed Mechanism
Yeast	Impaired degradation	Loss of Ufd1-Npl4-p97 retrotranslocation
Mammalian (RNAi)	Variable outcomes	Substrate-specific dependence on UFD1-p97 interaction

What advanced techniques elucidate UFD1’s role in DNA damage response (DDR) beyond ERAD?

Methodological Answer:
- Employ fluorescence microscopy to track UFD1-YFP/CFP-SUMO colocalization during Zeocin-induced DNA damage .
- Perform epistasis analysis with DDR mutants (e.g., rad22Δ, rhp51Δ) to identify synthetic lethality .
- Quantify SUMO conjugate turnover via ubiquitin pulldown assays in ufd1Δ backgrounds .

How can UFD1’s therapeutic potential in MYC-driven cancers be experimentally assessed?

Methodological Answer:
- Use zebrafish ufd1 heterozygous models to study MYC-driven T-ALL apoptosis and proliferation .
- Treat xenografts with UFD1 inhibitors (e.g., disulfiram/CuET) and measure tumor volume suppression .
- Combine UFD1 inactivation with ER stressors (tunicamycin) to amplify PERK/CHOP-mediated apoptosis .

Model	Key Readout	Citation
Zebrafish T-ALL	Caspase-3 activation, cell cycle arrest (G0/G1 phase)
MDA-MB-231 xenografts	Tumor volume reduction (77% with CuET)

What strategies address UFD1’s dual role in pro-survival ERAD and pro-apoptotic UPR?

Methodological Answer:
- Time-course experiments: Acute vs. prolonged ER stress (e.g., tunicamycin treatment at 0–24 hrs vs. >24 hrs) .
- Modulate UPR branches (PERK, IRE1, ATF6) using pharmacological inhibitors to isolate UFD1’s apoptotic effects .
- Transcriptomic profiling of UFD1−/− cells to identify MYC-regulated ER stress survival genes .

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ufd-1 Antibody