MED25 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- MED25 links COI1 with HAC1-dependent H3K9 acetylation to activate MYC2-regulated transcription of jasmonic acid-responsive genes. PMID: 28973940
- Plant density plays a crucial role in phase transitions, productivity, and root system architecture, highlighting the role of Mediator in self-plant recognition. PMID: 28556372
- MED25 is essential for papillae formation on the cell wall surface of leaf trichomes, suggesting that the Arabidopsis MED25 mediator component is likely involved in the transcription of a subset of genes that promote papillae deposition in trichomes. PMID: 28889249
- The PFT1 short tandem repeat is constrained to its approximate wild-type length by its various functional requirements. PMID: 25116137
- PHYTOCHROME AND FLOWERING TIME1 (PFT1), also known as MED25, is degraded by the proteasome, and proteasome-mediated PFT1 turnover is linked to its role in stimulating the transcription of FLOWERING LOCUS T. PMID: 22992513
- ROS homeostasis is crucial throughout the development of root hairs, controlling various processes via PFT1-regulated transcription of genes encoding redox-active enzymes. PMID: 23455023
- PFT1 regulates root hair growth through redox processes. PMID: 23106228
- The MED25 subunit of the Arabidopsis Mediator regulates a broad range of signaling pathways through selective interactions with specific transcription factors. PMID: 22822206
- PFT1 interferes with both positively and negatively acting components that regulate Arabidopsis light signaling. PMID: 22760208
- MED25 is required for the transcription activation ability of MYC2, ERF1, and ORA59. PMID: 22822211
- Genetic and physiological data suggest that MED25 (PFT1) functions to limit cell and organ growth independently of MED25-mediated phytochrome signaling and the jasmonate pathway. PMID: 21903673
- Med25 regulates PDF1.2 expression through these three transcription factors. PMID: 21343311
- Med25, in conjunction with drought response element binding protein 2A, participates in repressing PhyB-mediated light signaling, thus integrating signals from different regulatory pathways. PMID: 21536906
- Research has identified the proteins STRUWWELPETER (SWP) and PHYTOCHROME AND FLOWERING TIME 1 (PFT1) as the Med14 and Med25 subunits, respectively. PMID: 17560376
- Findings indicate that mediator complex subunit PFT1 is a key regulator of the jasmonate signaling pathway within the transcriptional machinery. PMID: 19671879
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with MED25 antibodies, incorporating methodological insights and data from diverse sources:
How do I validate the specificity of MED25 antibodies in Western blot (WB) and immunofluorescence (IF)?
Methodological Answer:
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Positive/Negative Controls: Use lysates from tissues/cell lines with confirmed MED25 expression (e.g., human spleen, brain) and knockout models .
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Blocking Peptide Assays: Pre-incubate antibodies with recombinant MED25 immunogen to confirm signal loss .
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Cross-Reactivity Checks: Validate across species (human, mouse, rat) using comparative WB .
| Validation Parameter | Recommended Approach | Source |
|---|---|---|
| WB Specificity | Lysates from HEK293 (high MED25) | |
| IF Specificity | Knockout cell lines + siRNA | |
| Species Reactivity | Parallel testing in human/mouse |
What are the optimal dilutions and conditions for MED25 antibody applications?
Methodological Answer:
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Western Blot: Start with 1:500–1:2000 dilution in 5% non-fat milk .
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IF/ICC: Use 1:50–1:200 dilution with Triton X-100 permeabilization .
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Fixation: 4% paraformaldehyde for 15 min, followed by antigen retrieval in citrate buffer (pH 6.0) .
How should MED25 antibodies be stored to ensure long-term stability?
How does MED25 regulate transcriptional coactivation in jasmonate (JA) signaling?
Methodological Answer:
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Interaction Studies: Use co-immunoprecipitation (co-IP) with MYC2 or ABI5 transcription factors in Arabidopsis .
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Chromatin Studies: Sequential ChIP-qPCR to detect MED25 and COI1 enrichment on JAZ8 or ERF1 promoters .
| Target Pathway | Key Partners | Technique | Source |
|---|---|---|---|
| JA Signaling | MYC2, COI1 | Co-IP + ChIP-qPCR | |
| ABA Signaling | ABI5 | Y2H + Luciferase |
How to resolve contradictory data on MED25’s role in ABA-responsive gene repression?
Methodological Answer:
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Domain Mapping: Test MED25 truncations (e.g., ACID domain) in yeast two-hybrid (Y2H) assays with ABI5 .
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Mutant Analysis: Compare med25-4 and myc2-2 mutants using qRT-PCR for PDF1.2 and ERF1 expression under ABA/JA treatments .
| Observation | Resolution Strategy | Source |
|---|---|---|
| MED25 represses ABA-responsive genes | Use med25-4 mutants + ABI5 co-IP |
What methods detect MED25 isoforms in mammalian systems?
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Isoform-Specific WB: Use NuPAGE gels (4–12% Bis-Tris) to resolve ~75–80 kDa bands .
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siRNA Knockdown: Target exon junctions unique to specific isoforms (e.g., isoforms 1–5) .
Data Contradiction Analysis
Example Issue: Discrepancies in MED25’s role in JA vs. ABA pathways.
Resolution Workflow:
