FTSH1 Antibody
Description
Immunogen and Epitope
The FTSH1 antibody is generated using a synthetic peptide derived from the Arabidopsis thaliana FTSH1 sequence (AT1G50250, UniProt ID: Q39102). This peptide is homologous to FTSH5 (AT5G42270), enabling cross-reactivity with related FtsH proteins in diverse plant species .
Techniques and Purposes
The FTSH1 antibody is primarily used in:
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Western Blotting: Detects FTSH1 in chloroplast membrane fractions, confirming its localization .
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Immunoprecipitation: Identifies protein interactions, such as the ubiquitination of FTSH1 by AtCHIP (an E3 ligase) in Arabidopsis .
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Immunolocalization: Maps FTSH1 to thylakoid membranes in chloroplasts .
Species Cross-Reactivity
The FTSH1 antibody demonstrates broad specificity across plant species and algae due to conserved FtsH domains:
| Species | Homology | Reference |
|---|---|---|
| Arabidopsis thaliana | 100% (target) | |
| Sorghum bicolor | High | |
| Chlamydomonas reinhardtii | Moderate | |
| Gossypium raimondii | High | |
| Nicotiana tabacum | Moderate |
Key Studies
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Protein Quality Control in Chloroplasts
FTSH1 is essential for degrading photo-damaged D1 protein in photosystem II (PSII). Antibody-based studies confirm its role in maintaining photosynthetic activity . -
Interaction with AtCHIP
Co-immunoprecipitation experiments reveal that AtCHIP binds FTSH1 in the cytoplasm, targeting its precursor for ubiquitination. Western blots using the FTSH1 antibody detected the mature 72 kDa form in thylakoid membranes . -
Cross-Species Functional Studies
While primarily used in Arabidopsis, the antibody’s cross-reactivity with FtsH5 in Sorghum and Gossypium suggests conserved roles in plant plastid maintenance .
Role in Chloroplast Development
FTSH1 is critical for:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research findings highlight the significance of protein activity and abundance in the functional integrity of FtsH heterocomplexes, which are composed of various isoforms within the thylakoid membrane. PMID: 21062893
- In darkness, FtsH hexamers are positioned adjacent to PSII complexes in the grana, enabling the prompt degradation of the photodamaged D1 protein under light stress. PMID: 20921219
Q&A
Here’s a structured FAQ for researchers working with FTSH1 antibodies, organized by scientific depth and methodological focus, based on current research findings:
What is the functional role of FTSH1 in apicomplexan parasites, and how can antibodies aid in studying it?
FTSH1 is an ATP-dependent metalloprotease critical for apicoplast biogenesis in Plasmodium falciparum and Toxoplasma gondii. Antibodies enable:
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Localization studies: Immunofluorescence to track apicoplast dynamics during parasite replication .
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Protein quantification: Western blotting to measure FTSH1 expression under drug treatment (e.g., actinonin) .
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Knockdown validation: Confirm reduced FTSH1 levels in conditional knockdown strains .
How do I validate FTSH1 antibody specificity for parasite vs. host homologs?
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Cross-reactivity testing: Use lysates from uninfected host cells (e.g., human hepatocytes) and parasite cultures in Western blotting .
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Epitope mapping: Compare antibody binding to conserved vs. divergent regions between host (YME1L1) and parasite FTSH1 (e.g., PfFtsH1 residues 91–612) .
Which experimental models are optimal for studying FTSH1 function?
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Parasite lines: Use P. falciparum 3D7 or Dd2 strains with inducible FTSH1 knockdown .
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Biochemical assays: Recombinant PfFtsH1 (ΔTM domain) for ATPase/protease activity profiling .
How to resolve contradictions between biochemical inhibition and phenotypic data in FTSH1 studies?
What strategies improve FTSH1 inhibitor selectivity over host metalloproteases?
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Structural analysis: Target non-conserved regions (e.g., PfFtsH1-specific loops) using cryo-EM or homology modeling .
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Analog testing: Replace actinonin’s hydroxamate group with bioisosteres (e.g., actinamide reduces potency 10-fold but retains specificity) .
How to address discrepancies between in vitro and in vivo FTSH1 inhibition?
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Pharmacokinetic profiling: Adjust drug exposure times to account for delayed apicoplast biogenesis defects .
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Metabolic labeling: Track apicoplast-targeted proteins (e.g., ACP) via radiolabeling post-inhibition .
Methodological Best Practices
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Antibody applications:
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Data normalization: Compare FTSH1 levels to apicoplast markers (e.g., ClpP1) to control for organelle loss .
