TOM20-4 Antibody
Description
Overview of TOM20 Antibodies
TOM20 antibodies are tools for studying mitochondrial protein import machinery, mitophagy, and diseases linked to mitochondrial dysfunction. These antibodies target TOM20, a 16–16.3 kDa receptor subunit of the TOM complex that recognizes nuclear-encoded mitochondrial precursor proteins .
Key Antibody Features
Tom20 (D8T4N) Rabbit mAb (#42406)
-
Reactivity: Human, Mouse, Rat, Monkey
-
Applications: WB, IP, IHC, IF
-
Key Data:
TOM20 Polyclonal Antibody (CAB21722)
-
Reactivity: Human, Mouse
-
Applications: WB, IF
-
Key Data:
Tom20 Antibody (F-10)
-
Host: Mouse IgG2a
-
Conjugates: HRP, PE, FITC, Alexa Fluor®
-
Key Data:
Mitophagy and Neurodegeneration
-
Mitophagy Assay: TOM20 antibodies quantify mitochondrial clearance in neuronal models (e.g., ReNcell VM cells). Co-treatment with oligomycin/antimycin (O/A) reduces TOM20 immunoreactivity, indicating mitophagy activation .
-
Parkinson’s Disease: TOM20 overexpression rescues α-synuclein-induced mitochondrial dysfunction, preserving dopaminergic neurons by restoring electron transport chain proteins .
Recommended Dilutions
| Antibody | WB | IHC | IF/ICC | IP |
|---|---|---|---|---|
| Proteintech 11802-1-AP | 1:5,000–50,000 | 1:500–2,000 | 1:50–500 | 0.5–4 µg/mg lysate |
| Tom20 (D8T4N) Rabbit mAb | 1:1,000 | 1:50 | 1:100 | 1:50 |
Cross-Reactivity
Disease Relevance
-
Neurodegeneration: Reduced TOM20 levels correlate with mitophagy defects in Parkinson’s and ALS .
-
Cancer: TOM20 overexpression in melanoma regulates Bax translocation, influencing pyroptosis .
Validation and Citations
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Basic Research Questions
How to validate TOM20-4 antibody specificity for mitochondrial outer membrane protein studies?
-
Perform Blue Native PAGE (BN-PAGE) with isolated mitochondria, followed by immunoblotting using TOM20-4 antibody. Compare migration patterns to known TOM complex subunits (e.g., TOM40 at 34 kDa).
-
Validate via protease susceptibility assays: Treat intact mitochondria with trypsin before lysis. TOM20-4 signal should disappear due to cytoplasmic domain degradation, while TOM40 remains intact .
-
Use knockout cell lines or siRNA-mediated TOM20 knockdown as negative controls .
What experimental models are appropriate for studying TOM20-4 dynamics?
-
HeLa cells: For single-molecule tracking of TOM20-4 diffusion dynamics using fluorescent protein (FP) tags (e.g., EGFP-Tom20) .
-
ReNcell VM neuronal models: To assess mitophagy via TOM20 immunostaining intensity under mitochondrial stressors (e.g., oligomycin/antimycin) .
-
Plant mitochondria: Arabidopsis outer membrane preparations for studying isoform-specific interactions (Table II) .
Advanced Research Questions
How to resolve contradictions in TOM20-4 molecular weight observations across studies?
-
Always include reducing/non-reducing Western blots to detect oxidation-induced dimerization, as cysteine residues (Cys13/Cys21) alter migration .
How to design assays differentiating TOM20-4’s receptor function from structural roles?
-
Proximity labeling: Use TOMM20-APEX2 constructs to map interactomes under varying metabolic states (e.g., iron stimulation). Compare partners like SYNJBP2 or PABPC4L against negative controls (Mito-APEX2) .
-
Functional rescue experiments: Express TOM20-4 mutants (e.g., C13S/C21S) in knockdown models. Monitor Bax translocation efficiency and caspase activation .
What methodologies address TOM20-4 antibody cross-reactivity in multi-species studies?
-
Epitope mapping: The D8T4N antibody targets a conserved N-terminal region (aa 50–70) across Homo sapiens, Mus musculus, and Arabidopsis .
-
Validate using:
Methodological Optimization
How to standardize TOM20-4 quantification in high-content mitophagy assays?
-
ReNcell VM protocol:
-
Normalize to nuclear counts (DAPI) to exclude cytotoxicity confounders.
How to interpret variable TOM20-4 diffusion coefficients in live-cell imaging?
-
Single-particle tracking (SPT) in HeLa cells reveals two subpopulations:
-
Use photoactivatable GFP tags to minimize overexpression artifacts.
Data Conflict Analysis
How to reconcile discrepancies in TOM20-4 interactome studies?
-
Combine orthogonal approaches: Crosslink with formaldehyde before BN-PAGE to stabilize weak interactions.
Why do TOM20-4 oxidation states affect antibody binding in stress models?
-
Under ROS (e.g., FeSO4/CCCP), Cys13/Cys21 form disulfide bonds, altering epitope accessibility .
-
Non-reducing SDS-PAGE + Western blot confirms oxidation. Always include β-mercaptoethanol controls.
Table: Arabidopsis TOM20 Isoforms
| Isoform | AA Length | Calculated MW (kDa) | pI |
|---|---|---|---|
| TOM20-4 | 187 | 21.0 | 5.4 |
