ced-9 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- HBx interacts directly with CED-9, a B-cell lymphoma 2 (Bcl-2) homolog, through a Bcl-2 homology 3 (BH3)-like motif. This interaction triggers both cytosolic Ca(2+) increase and cell death. PMID: 23091037
- EGL-1 converts CED-9 into a mitochondrial receptor for DRP-1, shifting its activity from mitochondrial fusion to fission. PMID: 21949250
- CED-9 retains the ability of mammalian Bcl-2 proteins to associate with cellular membranes, exhibiting amphitropic behavior. PMID: 21031486
- CED-9 forms a 2:2 heterotetrameric complex with CED-4 until specifically displaced by Caenorhabditis elegans BH3-only proteins, but not mammalian ones. PMID: 16167070
- CED-9 interacts with Mitofusin-2/fuzzy onions and can promote mitochondrial clustering and dramatic reorganization of mitochondrial networks. PMID: 16543146
- These findings support a multimodal character of CED-9 action, demonstrating its ability to regulate apoptosis through interactions in the cytosol and additional evolutionarily conserved roles at the membrane. PMID: 17703231
- While bcl-2 proteins are not required for ceramide to form protein-permeable channels in mitochondrial outer membranes, both recombinant human Bcl-x(L) and CED-9 disassemble ceramide channels in the mitochondrial outer membranes from rat liver and yeast. PMID: 18171672
- CED-9 can regulate the mitochondrial fission-fusion cycle but is not essential for either fission or fusion. PMID: 18827010
- CED-9 promotes Al tolerance in plants by inhibiting Al-induced programmed cell death. PMID: 19341713
Q&A
Basic Research Questions
What experimental approaches validate CED-9 antibody specificity in apoptosis studies?
CED-9 antibody specificity is typically confirmed using:
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Genetic knockout controls: Compare staining in wild-type vs. ced-9 loss-of-function mutants .
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Co-localization assays: Verify mitochondrial localization using markers like ATP synthase or cytochrome c .
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Western blot cross-reactivity tests: Ensure no binding to homologous BCL-2 family proteins (e.g., human BCL-2) .
How does CED-9 antibody aid in distinguishing its anti-apoptotic vs. pro-apoptotic roles?
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Subcellular fractionation: Separate mitochondrial vs. cytoplasmic CED-9 populations to study compartment-specific interactions .
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Co-immunoprecipitation (Co-IP): Identify binding partners (e.g., CED-4 or DRP-1) under apoptotic vs. non-apoptotic conditions .
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Mutation-based epitope mapping: Use CRISPR-edited ced-9 variants (e.g., CED-4-binding domain mutants) to validate functional domains .
Advanced Research Challenges
How to resolve contradictory data on CED-9’s dual roles in apoptosis regulation?
What methodologies address CED-9’s interaction with mitochondrial fission/fusion proteins?
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GST pull-down assays: Confirm direct binding between CED-9 and DRP-1 (e.g., Fig. 1A-B in ).
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Live-cell imaging: Track mitochondrial dynamics in ced-9 RNAi-treated worms .
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Quantitative mass spectrometry: Identify phosphorylation sites regulating CED-9/DRP-1 binding .
How to design experiments isolating CED-9’s anti-apoptotic function from its pro-death role?
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Conditional overexpression: Use heat-shock promoters (e.g., P<sub>HS</sub> ced-9) to temporally control expression .
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Tissue-specific knockdown: Combine CED-9 antibody staining with cell-type-specific reporters (e.g., pharyngeal vs. neuronal cells) .
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Bimolecular fluorescence complementation (BiFC): Visualize real-time CED-9/CED-4 complexes during apoptosis initiation .
Mechanistic & Translational Questions
What evidence supports CED-9’s conserved role in mammalian apoptosis pathways?
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Functional complementation: Human BCL-2 rescues ced-9 mutants but retains anti-apoptotic activity only .
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Cross-species Co-IP: CED-9 antibody detects interactions with mammalian APAF-1 in transgenic models .
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Structural modeling: Predict conserved binding interfaces using AlphaFold2 (e.g., BH3 domain alignment) .
How to troubleshoot CED-9 antibody cross-reactivity in mutant screens?
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Epitope tagging: Fuse CED-9 with HA/FLAG tags for dual antibody validation .
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Competitive ELISA: Pre-incubate antibody with recombinant CED-9 protein to block non-specific binding .
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Single-cell RNA-FISH: Correlate ced-9 mRNA levels with protein detection in apoptotic cells .
