anti-Homo sapiens (Human) UBE2U Antibody raised in Rabbit

Description

Introduction to UBE2U and Its Role in Cellular Processes

UBE2U (Ubiquitin Conjugating Enzyme E2 U) is an E2 ubiquitin-conjugating enzyme that facilitates the transfer of ubiquitin to substrate proteins, a critical step in protein ubiquitination. This post-translational modification regulates diverse cellular processes, including DNA damage response (DDR), cell cycle checkpoints, and chromatin remodeling . UBE2U operates in tandem with E3 ligases like RNF17 to enforce DDR by promoting the accumulation of repair proteins (e.g., 53BP1, BRCA1) at double-strand breaks (DSBs) . Its catalytic activity is essential for maintaining genome stability and mitigating genotoxic stress .

Development and Validation of UBE2U Antibodies

UBE2U antibodies are critical tools for studying its expression, localization, and mechanistic roles. Key challenges include detecting endogenous UBE2U due to low abundance, necessitating highly specific antibodies . Validated antibodies include:

Provider	Catalog Number	Clonality	Applications
Proteintech Group	17323-1-AP	Polyclonal	WB, ELISA, ICC, IHC
LSBio	LS-C409912	Polyclonal	WB, IHC
NovoPro Bioscience Inc.	116538	Polyclonal	WB, ELISA, ICC, IHC

These antibodies have been employed in Western blot (WB), immunohistochemistry (IHC), and immunocytochemistry (ICC) to study UBE2U in cancer models and DDR pathways .

DNA Damage Response Regulation

RNAi Screening: UBE2U depletion via siRNA impaired 53BP1 and BRCA1 focus formation at DSBs, highlighting its role in chromatin remodeling post-irradiation .
Checkpoint Control: UBE2U-deficient cells exhibited defective G2/M checkpoint arrest, linking its activity to cell cycle regulation .

Interaction With E3 Ligases

UBE2U partners with RNF17, an E3 ligase, to regulate RNF168-dependent ubiquitination at DSBs. This interaction was confirmed via co-immunoprecipitation and siRNA-mediated knockdown experiments .

Challenges and Considerations in UBE2U Antibody Usage

Low Endogenous Detection: Most antibodies (e.g., Novus H00148581-B01) perform well with overexpressed UBE2U but struggle with endogenous levels .
Specificity: Cross-reactivity with other E2 enzymes (e.g., UBE2L3) requires rigorous validation .

Product Specs

Buffer

The antibody is provided in phosphate buffered saline (PBS) containing 0.1% sodium azide, 50% glycerol, and adjusted to pH 7.3. Store at -20°C. Avoid repeated freeze-thaw cycles.

Lead Time

We typically dispatch orders within 1-3 business days of receipt. Delivery times may vary depending on the method of purchase and location. Please contact your local distributor for specific delivery timelines.

Synonyms

RP4-636O23.1 antibody; UBE2U antibody; UBE2U_HUMAN antibody; Ubiquitin carrier protein U antibody; Ubiquitin conjugating enzyme E2U (putative) antibody; Ubiquitin-conjugating enzyme E2 U antibody; Ubiquitin-protein ligase U antibody

Target Names

UBE2U

Uniprot No.

Q5VVX9

Target Background

Function

UBE2U Antibody catalyzes the covalent attachment of ubiquitin to other proteins.

Database Links

HGNC: 28559

KEGG: hsa:148581

STRING: 9606.ENSP00000360116

UniGene: Hs.388746

Protein Families

Ubiquitin-conjugating enzyme family

Q&A

FAQs for UBE2U Antibody in Academic Research

Advanced Research Questions

How to resolve contradictions in UBE2U’s reported roles in K48 vs. K63 ubiquitination?

Methodological considerations:

Approach	Purpose
In vitro reconstitution with purified E1, UBE2U, and E3 ligases	Define intrinsic chain-type preference
Mutagenesis of UBE2U’s catalytic cysteine (C87A)	Confirm enzyme dependency
Linkage-specific ubiquitin antibodies (e.g., K48 vs. K63)	Distinguish chain types

Analyze tissue-specific effects: Muscle vs. neuronal systems show divergent E2-substrate preferences .

What strategies mitigate off-target effects in UBE2U antibody-based proximity ligation assays (PLA)?
- Pre-block membranes with lysates from UBE2U−/− cells to reduce non-specific binding .
- Combine PLA with Förster resonance energy transfer (FRET) using fluorescently tagged ubiquitin .
- Validate interactions via orthogonal methods (e.g., crosslinking-MS) .

Technical Challenges & Solutions

How to distinguish UBE2U’s role in canonical vs. non-canonical ubiquitination pathways?
- Experimental design:
  1. Use diGly remnant enrichment proteomics to identify UBE2U-specific substrates.
  2. Compare ubiquitination profiles in UBE2U knockdown vs. wild-type cells under proteasome inhibition (MG132) .
  3. Employ linkage-specific deubiquitinases (DUBs) to cleave specific chain types .
What controls are critical for UBE2U antibody-based subcellular localization studies?
- Required controls:
  - Co-staining with organelle markers (e.g., LAMP1 for lysosomes, Calnexin for ER) .
  - Pharmacological disruption (e.g., Bafilomycin A1 for lysosomal inhibition) .
  - Fractionation-Western blotting to confirm antibody specificity across compartments .

Data Interpretation Guidelines

How to address discrepancies in UBE2U expression levels across disease models?

Analysis framework:

Factor	Consideration
Tissue heterogeneity	Use laser-capture microdissection for precise sampling
Post-translational modifications	Treat lysates with phosphatases/proteases
Antibody lot variability	Include reference standards in all blots

What bioinformatics tools enhance UBE2U substrate prediction?
- Integrate UbPred and UbiNet databases with experimental IP-MS data .
- Use structural modeling (AlphaFold) to predict UBE2U-E3 interfaces .
- Apply machine learning (Scikit-learn) to identify sequence motifs in validated substrates .

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UBE2U Antibody