RPP3B Antibody
Description
Ribosomal Protein L3 (RPL3) Antibodies
RPL3 is a core component of the 60S ribosomal subunit involved in translation elongation. Two well-characterized antibodies include:
Key Findings:
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WB Validation: Both antibodies detect endogenous RPL3 at 46 kDa in multiple cell lines (HEK-293, RAW 264.7) .
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IHC Performance: Strong cytoplasmic staining in human cancer tissues (liver, breast, lung) with antigen retrieval .
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Subcellular Localization: Predominantly cytoplasmic in HepG2 cells via immunofluorescence .
Ribosomal Protein S3 (RPS3) Antibodies
RPS3 is a 27 kDa component of the 40S ribosomal subunit with roles in DNA repair and apoptosis.
Key Findings:
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Western Blot: Detects RPS3 at 27 kDa in HepG2, PC-12, and Neuro-2A lysates .
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Immunofluorescence: Localizes to cytoplasmic ribosomes in Neuro-2A cells .
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Clinical Relevance: Used to study ribosome biogenesis defects in cancer models .
Antibody Validation Metrics
Recent studies emphasize rigorous validation for ribosomal protein antibodies:
Table 3: Validation Standards
Research Applications
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Cancer Biology: Used to quantify RPL3/RPS3 overexpression in hepatocellular carcinoma .
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Neurological Studies: Neuro-2A cell models employ these antibodies to study ribosome stress .
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Structural Biology: Supports cryo-EM studies of ribosomal complexes .
Challenges in Antibody Reliability
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Batch Variability: 23% of commercial ribosomal antibodies show inter-lot inconsistency .
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Epitope Mapping: Only 34% of antibodies targeting ribosomal proteins have mapped linear epitopes .
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Phosphorylation Status: Most commercial antibodies do not distinguish phosphorylation states (e.g., RPS3 Ser6) .
Emerging Alternatives
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Transgenic sweetpotato plants expressing the Arabidopsis ribosomal P3 (AtP3B) gene under the control of the CaMV 35S promoter (referred to as OP plants) were generated. Three OP lines exhibited enhanced heat tolerance, increased photosynthesis efficiency, and improved low temperature tolerance compared to wild type plants. PMID: 28806972
- Research indicates that RPP3B plays a protective role in cells against both high- and low-temperature stresses. PMID: 27004478
Q&A
FAQs for Researchers on UPF3B/RENT3B Antibody in Academic Research
Advanced Research Questions
How to resolve contradictions in UPF3B expression data across studies?
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Analytical framework:
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Cross-platform validation: Compare RPPA results using clustered antibody-antigen maps to identify concordant/discordant signals (Fig. 1) .
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Epitope mapping: Confirm antibody binding regions (e.g., UPF3B aa 300–350) to rule out isoform-specific artifacts .
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Batch variability assessment: Replicate experiments with independent antibody lots to isolate technical vs. biological variability .
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Table 1: Antibody Concordance Across RPPA Platforms
How to design multiplex assays incorporating UPF3B antibody?
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Integrated workflow:
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Pair UPF3B WB/IP with RNA-seq to correlate nonsense-mediated decay (NMD) activity with mRNA stability .
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Combine with phospho-specific antibodies (e.g., UPF1 phospho-sites) to study post-translational regulation of NMD .
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Use machine learning to cluster UPF3B interaction partners identified via mass spectrometry .
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How to address UPF3B’s dual role in translation stimulation and NMD?
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Functional dissection:
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Domain-specific mutagenesis: Delete UPF3B’s EJC-binding domain to isolate translation vs. NMD roles .
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Ribo-seq: Compare ribosomal occupancy in UPF3B-knockout vs. wild-type cells under stress conditions .
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Ternary complex assays: Use EMSA to test UPF3B’s interaction with UPF2 and RNA substrates in vitro .
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Methodological Best Practices
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Critical controls:
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Data interpretation:
