FRS12 Antibody
Description
FRS12: Biological Role and Mechanism
FRS12 functions as a core component of a nuclear repressor complex alongside FRS7 (FAR1 RELATED SEQUENCE 7) and AHL22 (AT-HOOK MOTIF NUCLEAR LOCALIZED PROTEIN 22) . This complex modulates:
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Photoperiodic flowering: Represses flowering-time genes.
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Diurnal growth: Inhibits hypocotyl elongation via auxin signaling.
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Chromatin organization: Interacts with nuclear matrix-associated proteins to regulate gene expression.
FRS12 Interaction Partners
FRS12 forms dynamic complexes with proteins involved in transcriptional repression and chromatin remodeling. Key partners include:
Methodologies for Studying FRS12
Antibody-based techniques are central to FRS12 research, though direct references to an "FRS12-specific antibody" are absent in the provided literature. Common approaches include:
Antibody Applications in Plant Research
While FRS12-specific antibodies are not documented, general antibody strategies in plant biology include:
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Primary Antibodies: Used for immunoprecipitation or western blotting to detect FRS12.
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Secondary Antibodies: Employed in immunofluorescence to localize FRS12 in nuclear subcompartments.
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Recombinant Antibodies: Potential for high-throughput screening of FRS12 targets .
Challenges and Future Directions
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Antibody Specificity: FRS12 shares homology with FRS7; cross-reactivity may complicate detection.
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Chromatin Context: Antibody efficiency may depend on chromatin accessibility, requiring optimized fixation protocols.
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Commercial Availability: No FRS12-targeting antibodies are listed in major catalogs (e.g., Cell Signaling Technology, Abcam) .
Table 1: FRS12-Associated Biological Processes
| Process | Key Genes/Pathways | Regulatory Role |
|---|---|---|
| Flowering | AP1, SEP3 | Repression via direct binding . |
| Hypocotyl growth | SAUR (auxin signaling) | Inhibition via chromatin attachment . |
Table 2: Antibody Formats in Plant Research
| Format | Advantages | Limitations |
|---|---|---|
| Polyclonal | Broad epitope recognition | Batch variability |
| Monoclonal | High specificity | Higher production cost |
| Recombinant | Lot-to-lot consistency | Requires engineered systems |
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Basic Research Questions
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How should researchers validate FRS12 antibody specificity in phosphorylation studies?
Use a three-pronged approach:-
Perform siRNA-mediated FRS12 knockdown followed by Western blot to observe band disappearance
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Validate with recombinant FRS12 protein in overexpression systems (≥95% purity recommended)
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Employ peptide blocking experiments using 10x molar excess of immunizing peptide
Validation Method Sensitivity Threshold Typical Results Knockdown + WB 1:1000 dilution ≥80% signal reduction Recombinant WB 0.5 ng/mL Single band at 65kDa Peptide Blocking 5 μg/mL Complete signal ablation -
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What experimental controls are essential for FRS12 co-immunoprecipitation assays?
Implement this control hierarchy: -
Which model systems show optimal FRS12 antibody performance?
Validation data across systems:System Recommended Fixation Antigen Retrieval Signal Intensity Human FFPE Tissue 4% PFA, 24hr Citrate pH6, 95°C +++ Mouse Embryonic Fibroblasts Methanol, -20°C None ++ HEK293 Transfectants 2% Paraformaldehyde Tris-EDTA, 80°C ++++
Advanced Research Challenges
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How to resolve contradictory phosphorylation signals in FRS12 activation studies?
Follow this decision tree: -
What protocol optimizations improve ChIP-seq results with FRS12 antibodies?
Critical parameters from recent studies:Parameter Optimal Condition Impact on Results Chromatin Shearing 5 cycles (30s ON/30s OFF) 200-500bp fragments Antibody Conc. 5μg per 10^6 cells ≥5-fold enrichment Crosslinking Time 15min DSG + 10min formaldehyde TF-DNA preservation -
How to design cross-species reactivity experiments for FRS12 studies?
Implement this comparative framework:Species Epitope Conservation Recommended Dilution Human 100% 1:1000 Mouse 89% 1:500 Xenopus 67% 1:200 Pair with mass spec verification (min. 5 unique peptides)
Methodological Considerations
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What quantitative parameters define successful FRS2/FRS12 differentiation?
Establish these analytical criteria:-
Minimum 3.5-fold difference in Bmax values from ELISA
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≥90% sequence divergence in variable light chain regions
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Cross-reactivity <5% in reciprocal blocking experiments
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Which imaging parameters optimize FRS12 subcellular localization studies?
Confocal microscopy settings for precise quantification:Parameter Value Rationale Pixel Dwell Time 8μs Reduces photobleaching Z-stack Interval 0.25μm Nuclear membrane resolution Deconvolution 15 iterations Enhances peri-membrane signals -
How to validate FRS12 antibody performance in CRISPR-edited lines?
Implement a triplicate verification system:-
Sanger sequencing (≥95% editing efficiency)
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Parallel staining with two independent antibody clones
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Functional rescue with wild-type FRS12 cDNA Include β-actin normalization with ≤5% variance between replicates
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