urm-1 Antibody
Description
What is URM1 Antibody?
URM1 antibodies are immunological reagents designed to detect and quantify URM1, a ubiquitin-like protein (UBL) that functions as a sulfur carrier for tRNA modification and covalently conjugates to target proteins during oxidative stress . These antibodies are critical for:
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Identifying URM1-protein conjugates (urmylation) in stress response pathways .
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Studying URM1’s role in tRNA 2-thiolation, which optimizes ribosomal translation .
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Analyzing URM1’s association with cancer progression and apoptosis .
Validation Data for URM1 Antibodies
Key commercially available URM1 antibodies include:
Key Validation Results:
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ab110699: Detects recombinant URM1 at 0.01 µg and endogenous URM1 in HepG2 lysates .
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15285-1-AP: Validated in WB (human placenta, A549 cells) and IHC (mouse liver) .
tRNA Thiolation and Oxidative Stress
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URM1 antibodies confirm its thiocarboxylated form (-COSH), which donates sulfur for tRNA wobble uridine modification (mcm⁵s²U) .
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Oxidative stress (e.g., diamide, H₂O₂) induces URM1 conjugation to substrates like Ahp1 and USP15, detectable via WB .
Cytokinesis and Cell Cycle Regulation
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Knockdown studies using URM1-targeting shRNA revealed cytokinesis defects in HeLa cells, including multinucleation and >4n DNA content .
Cancer Research
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Liver Cancer: High URM1 expression correlates with poor prognosis in hepatocellular carcinoma (HCC). URM1 knockdown suppresses proliferation, migration, and tumor growth in xenograft models .
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Toxoplasma gondii: URM1 deletion reduces parasite virulence and stress tolerance, highlighting its role in pathogen survival .
URM1 in Apoptosis and Signaling
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URM1 knockdown upregulates phosphorylated JNK1/2 and p53, promoting apoptosis in liver cancer cells .
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Table: URM1 Knockdown Effects in HCC (n=90)
Parameter URM1 High vs. Low P-value Tumor size Larger <0.01 Survival rate Reduced <0.05 Apoptosis markers Increased <0.01
Urmylation in Stress Response
Product Specs
Target Background
Q&A
Basic Research Questions
What experimental validation strategies ensure specificity of URM1 antibodies in Western blotting?
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Methodological approach:
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Perform siRNA/shRNA-mediated URM1 knockdown (≥95% reduction) in target cell lines (e.g., HeLa, U2OS) and compare band intensity between control and knockdown lysates .
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Use recombinant URM1 protein (11 kDa theoretical size) as a positive control to confirm antibody reactivity at the expected molecular weight .
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Validate cross-reactivity across species using lysates from human, mouse, and rat tissues, as URM1 is evolutionarily conserved .
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How do researchers select model organisms for URM1 functional studies?
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Key considerations:
Advanced Research Questions
How can URM1 antibody-based proteomics resolve contradictory data on its tRNA modification vs. ubiquitin-like roles?
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Integrated methodology:
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Functional proteomics: Use URM1 vinyl methylester (VME) probes to covalently trap interacting enzymes (e.g., ATPBD3, MOCS3) in HEK293T lysates, followed by LC-MS/MS .
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Oxidative stress assays: Perform Western blotting with anti-URM1 under diamide/H₂O₂ treatment to detect UBL-conjugates (e.g., Ahp1 modification) .
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tRNA analysis: Combine antibody-based immunoprecipitation with mass spectrometry to quantify 2-thiolation levels at mcm⁵S²U wobble positions .
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What controls are critical when interpreting URM1 immunohistochemistry (IHC) data in cancer studies?
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Technical safeguards:
How do researchers reconcile cell type-specific variability in URM1 phenotypes?
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Case analysis:
What multi-omics strategies leverage URM1 antibodies to study its dual roles?
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Workflow integration:
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Ubiquitinome profiling: Immunoprecipitate URM1 conjugates from diamide-treated cells, identify targets via LC-MS/MS (e.g., Ahp1, CAS) .
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tRNA sequencing: Compare ΔUrm1 vs. wild-type strains to quantify wobble uridine thiolation (requires ≥50× coverage) .
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Structural biology: Use antibody-guided crosslinking to map URM1’s interaction interface with MOCS3/ATPBD3 .
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Data Contradiction Analysis
Why do some studies report URM1 as essential while others show viability in knockout models?
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Resolution framework:
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Species differences: Human cells exhibit cytokinesis failure , while T. gondii survives with attenuated virulence .
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Stress conditions: URM1 is dispensable under standard culture but critical under oxidative stress (e.g., 1.5 mM diamide reduces ΔUrm1 survival by 70%) .
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Functional redundancy: Thioredoxin systems may compensate in certain cell types (e.g., DF-1 fibroblasts ).
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How to troubleshoot inconsistent URM1 conjugation signals in Western blots?
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Optimization checklist:
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Lysis buffer: Include 20 mM N-ethylmaleimide to preserve thioester bonds .
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Electrophoresis: Use 4–12% Bis-Tris gels to resolve URM1 (~11 kDa) from conjugates (e.g., URM1-Ahp1 at ~35 kDa) .
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Antibody validation: Compare multiple clones (e.g., Proteintech 15285-1-AP vs. Abcam EPR4809) across ≥3 biological replicates .
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