RDM16 Antibody
Description
PRDM16 Antibody Overview
PRDM16 (PR Domain-Containing Protein 16) is a zinc finger transcription factor involved in:
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Brown vs. white adipose tissue differentiation (regulates thermogenesis and energy metabolism) .
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TGF-β signaling repression (impacts orofacial development and cancer) .
Antibodies targeting PRDM16 are essential for studying its role in adipose tissue biology, oncogenesis, and developmental processes.
Applications and Detection Methods
PRDM16 antibodies are validated for diverse techniques:
Note: Molecular weight discrepancies arise from isoforms (e.g., isoform 4 lacks the PR domain ) or post-translational modifications.
Research Findings and Functional Insights
PRDM16 antibodies have elucidated critical roles in:
Adipose Tissue and Metabolism
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Brown fat biogenesis: PRDM16 stabilizes beige fat through post-translational modifications (e.g., phosphorylation, sumoylation) .
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White fat suppression: PRDM16 represses white adipose tissue differentiation via CEBPB cooperation .
Cancer and Development
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Myelodysplastic syndrome (MDS)/Acute Myeloid Leukemia (AML): Truncated PRDM16 (lacking the PR domain) is overexpressed in t(1;3)-positive MDS/AML cases .
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Brain development: PRDM16 interacts with LHX2 to specify retinal ganglion cell subtypes .
Signaling Pathways
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TGF-β repression: PRDM16 binds Smad3, inhibiting TGF-β-driven gene expression .
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Histone methylation: PRDM16 monomethylates H3K9, influencing chromatin structure and gene regulation .
Challenges and Recommendations
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Isoform variability: PRDM16 isoforms (e.g., isoform 4) may alter antibody specificity .
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Sample types: Validate antibodies in nuclear extracts (brown adipose tissue) or frozen sections (embryos) .
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Signal enhancement: Use near-infrared fluorescence (e.g., Licor streptavidin 800) for low-abundance targets .
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs tailored for academic research on RDM16 antibodies, incorporating methodological insights and data from peer-reviewed studies:
What is RDM16’s functional role in plant heat stress responses?
RDM16 (RNA-DIRECTED DNA METHYLATION 16) is a pre-mRNA splicing factor critical for heat stress tolerance in Arabidopsis thaliana. It regulates the splicing of 18/20 HEAT SHOCK TRANSCRIPTION FACTOR (HSF) mRNAs, enabling proper heat stress responses. Knockout mutants (rdm16-1, rdm16-4) exhibit hypersensitivity to heat stress, impaired HSF splicing, and reduced survival rates under heat treatment .
Methodological Insight: To study RDM16’s role:
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Use RT-qPCR to quantify HSF splicing efficiency in mutants vs. wild-type plants.
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Perform transcriptomic profiling (RNA-seq) to identify differentially expressed genes (DEGs) under heat stress.
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Validate condensation behavior via GFP-tagged RDM16 in protoplasts under heat stress .
How do researchers validate RDM16 antibody specificity?
Validation involves:
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Western Blot: Detect a ~170 kDa band in lysates (e.g., K562 human leukemia cells) .
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Knockdown Validation: Compare RDM16 protein levels in wild-type vs. rdm16 mutants .
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Immunofluorescence: Localize RDM16 condensates in nuclei under heat stress (42°C) .
Data Table: Antibody Performance Across Studies
| Application | Host | Target Species | Key Validation Result | Source |
|---|---|---|---|---|
| Western Blot | Sheep | Human/Mouse | 170 kDa band in K562 lysates | |
| Immunofluorescence | Rabbit | Arabidopsis | Nuclear condensates at 42°C |
How do RDM16 splice variants (RDL/RDS) affect experimental outcomes?
RDM16 produces two splice isoforms:
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RDM16-LONG (RDL): Full-length protein with intact IDR1 (intrinsically disordered region).
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RDM16-SHORT (RDS): Lacks part of IDR1 but enhances RDL condensation under heat stress .
Methodological Insight:
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Design isoform-specific primers for RT-qPCR to distinguish RDL/RDS expression.
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Use GFP-tagged constructs to compare condensate formation between isoforms .
What are common pitfalls in detecting RDM16 in tissue-specific contexts?
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Cross-Reactivity: Commercial PRDM16 antibodies (e.g., AF6295) may cross-react with non-target proteins due to conserved domains .
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Epitope Masking: Heat-induced condensates may obscure antibody binding sites.
Workflow Optimization:
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Pre-treat samples with proteinase K to expose epitopes in condensates.
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Combine immunofluorescence with transcriptomic data to confirm functional relevance .
How to resolve discrepancies in RDM16 antibody performance across studies?
Case Example: A study reports a 170 kDa band , while another detects smaller isoforms .
Troubleshooting Strategy:
| Factor | Investigation Method | Source |
|---|---|---|
| Post-Translational Modifications | Phos-tag gel electrophoresis | |
| Species Specificity | Compare reactivity in Arabidopsis vs. human |
How to combine RDM16 antibody-based assays with transcriptomic data?
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Perform co-analysis of DEGs (e.g., HSA32, APX2, COR15A) and RDM16 protein levels under heat stress .
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Use ChIP-seq with anti-RDM16 antibodies to identify direct splicing targets.
Data Correlation Table:
| Gene | Expression (Log2FC) | Splicing Efficiency (WT vs. rdm16) |
|---|---|---|
| HSFA2 | +3.2 | 85% reduction in rdm16 |
| APX2 | +1.8 | 72% reduction |
