TIM23-2 Antibody
Product Specs
Target Background
Q&A
Here’s a structured FAQ collection for researchers studying TIM23-2 Antibody in mitochondrial biology, synthesized from peer-reviewed studies and technical methodologies:
What advanced techniques combine TIM23-2 antibody with functional assays for mitochondrial import studies?
Methodological Answer:
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Import Arrest Assays: Trap preproteins in the TIM23 channel using matrix-targeted substrates (e.g., b₂Δ construct) and analyze co-precipitated subunits via IP ( ).
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Electrophysiology Integration: Pair antibody-based immunoblotting with patch-clamp recordings to correlate TIM23 channel gating with complex composition (Fig. 1D, ).
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Pulse-Chase + IP: Monitor precursor protein translocation kinetics using radiolabeled preproteins and quantify TIM23-bound intermediates ( ).
How to resolve contradictions in TIM23 complex subunit associations across studies?
Methodological Answer:
How to optimize TIM23-2 antibody concentration for co-immunoprecipitation (CoIP)?
Methodological Answer:
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Titration Series: Test antibody dilutions (1:50–1:500) in digitonin-solubilized mitochondria to balance specificity and yield ( ).
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Negative Controls: Include preimmune IgG and Tim23-deficient mitochondria to exclude nonspecific binding ( ).
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Quantitative WB: Normalize CoIP eluates to input (e.g., anti-Cox4 for mitochondrial loading control; Fig. 5F, ).
What are limitations of TIM23-2 antibody in studying dynamic complex rearrangements?
Methodological Answer:
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Translocation-Induced Dissociation: Active import destabilizes Tim21/PAM interactions; use import-arrested preproteins to "freeze" intermediate states ( ).
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Membrane Potential Sensitivity: Δψ loss disrupts TIM23-precursor interactions; stabilize with CCCP during lysis ( ).
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Epitope Masking: Conformational changes during gating may reduce antibody accessibility; validate with multiple epitope tags (e.g., HIS/GST-Mgr2; ).
How to design experiments linking TIM23-2 antibody data to mitochondrial respiratory deficits?
Methodological Answer:
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OXPHOS Profiling: Correlate Tim23 levels (via WB) with OXPHOS-CR gene expression (e.g., ATP5L, COX5B; ).
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Functional Rescue: Overexpress wild-type Tim23 in knockdown models and assess import recovery using mtGFP processing assays ( ).
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Metabolic Flux Analysis: Combine Seahorse assays with Tim23 IP to identify interactome changes under energetic stress ( , ).
How does TIM23-2 antibody performance vary across species or tissue types?
Methodological Answer:
Advanced: What computational tools complement TIM23-2 antibody data for complex modeling?
Methodological Answer:
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Molecular Dynamics: Use Cryo-EM structures (e.g., Tim23-Tim17 interface) to predict antibody accessibility (TM2 mutations in ).
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Network Analysis: Apply STRING or BioGRID to map TIM23 interactors identified via IP-MS ( , ).
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Machine Learning: Train classifiers on OXPHOS-CR gene expression ( ) to predict TIM23 dysfunction from antibody quantification data.
