AGP10 Antibody
Description
Introduction to AGP10 Antibody
The term "AGP10 Antibody" appears to be a potential misnomer or typographical error in available literature. Based on comprehensive analysis of scientific sources, the correct designation is ATG10 Antibody, targeting the Autophagy Related 10 (ATG10) protein, a critical enzyme in autophagy—a cellular degradation pathway. ATG10 facilitates the conjugation of ATG12 to ATG5, a step essential for autophagosome formation . This antibody is widely used in autophagy research to study molecular mechanisms in cancer, neurodegenerative diseases, and immune regulation .
Role in Autophagy and Disease
ATG10 is indispensable for autophagosome maturation, linking ATG12 to ATG5 to form the ATG12–ATG5 conjugate, which promotes LC3 lipidation and membrane elongation . Dysregulation of ATG10 is implicated in:
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Cancer: Elevated ATG10 expression correlates with tumor progression and chemoresistance .
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Neurodegeneration: ATG10 deficiency disrupts protein aggregate clearance in Parkinson’s disease models .
Validation Studies
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Western Blot: Antibody A304840 shows specificity at 1:1,000 dilution in human cell lysates .
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Immunofluorescence: Demonstrated cytoplasmic localization in SK-N-BE neuroblastoma cells .
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Cross-Reactivity: Antibody DF8366 reacts with human, mouse, and rat tissues but not in AGP-deficient models .
Comparative Analysis of Antibody Performance
| Parameter | A304840 | ABIN2429199 | DF8366 |
|---|---|---|---|
| Sensitivity (WB) | 1:1,000 | 1:500 | 1:1,000 |
| Specificity | Human-specific | Multi-species | Multi-species |
| Peer-Reviewed Citations | 2+ | 5+ | 3+ |
Challenges and Limitations
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Epitope Accessibility: ATG10’s conformational flexibility may reduce antibody binding efficiency in fixed tissues .
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Cross-Reactivity: Some antibodies show non-specific binding to ATG10 homologs in non-target species .
Future Directions
Recent advances in structural biology (e.g., cryo-EM) could refine epitope mapping for ATG10 antibodies, enhancing specificity . Additionally, CRISPR-based validation in ATG10-knockout models is recommended to confirm antibody reliability .
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
What is the role of AGP10 in autophagy, and how do antibodies aid in its study?
AGP10 functions as an E2-like enzyme in two ubiquitin-like conjugation pathways: (1) covalent linkage of ATG12 to ATG5, forming a complex critical for autophagosome elongation, and (2) lipid modification of MAP-LC3, enabling its membrane association during autophagosome formation . Anti-AGP10 antibodies enable researchers to:
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Track protein expression via Western blot (WB) or immunofluorescence (IF), using epitopes such as the immunogen sequence RPLTLKDIWEGVHECYKMRLLQGPWDTITQQEHPILGQPFFVLHPCKTNEFMTPVLKNSQKINKNVNYITSWLSIVGPVVGLNLPLSYAKATSQDERNVP .
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Validate knockdown/knockout models by comparing AGP10 levels in wild-type versus genetically modified cells (e.g., CRISPR-Cas9 systems) .
Table 1: Common AGP10 Antibody Clones and Applications
| Clone | Applications | Species Reactivity | Key Validation Data | Source |
|---|---|---|---|---|
| ARC1425 | WB, IF | Human, Mouse | Reduced signal in ATG10-KO cells | Thermo Fisher |
| EPR22456-88 | sELISA | Mouse | Linear detection range: 0.1–10 ng/mL | Abcam |
How should researchers select and validate anti-AGP10 antibodies for Western blot?
Selection Criteria:
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Immunogen Specificity: Prioritize antibodies raised against full-length AGP10 or domains involved in ATG12/MAP-LC3 binding (e.g., residues 50–150) .
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Cross-Reactivity: Verify reactivity with target species using databases like UniProt (Human: Q9H0Y0; Mouse: Q8R1P4) .
Validation Steps:
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Knockout Controls: Use ATG10-deficient cell lines (e.g., HeLa ATG10-KO) to confirm antibody specificity .
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Dose-Response: Test antibody dilutions (1:500–1:2000) to optimize signal-to-noise ratios .
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Cross-Validation: Compare results with orthogonal methods (e.g., mRNA quantification via qPCR) .
What are common pitfalls in AGP10 antibody-based assays, and how can they be mitigated?
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Non-Specific Bands in WB: Caused by antibody cross-reactivity with paralogs (e.g., ATG7). Mitigation: Pre-adsorb antibodies with ATG7 knockout lysates .
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Variable IF Staining: Due to fixation artifacts. Solution: Compare paraformaldehyde (4%) vs. methanol fixation effects .
How can researchers resolve discrepancies in AGP10 subcellular localization studies?
Conflicting reports describe AGP10 as cytoplasmic or membrane-associated . To address this:
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Fractionation Assays: Separate cytosolic and membrane fractions using ultracentrifugation, then probe with anti-AGP10 antibodies .
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Live-Cell Imaging: Fuse AGP10 with fluorescent tags (e.g., GFP) and track dynamics under autophagy-inducing conditions (e.g., nutrient deprivation) .
Table 2: AGP10 Localization Under Autophagy-Inducing Conditions
| Condition | Localization | Antibody Clone | Supporting Evidence |
|---|---|---|---|
| Basal state | Cytoplasmic | ARC1425 | Co-staining with cytosolic markers |
| Starvation (6 hr) | Pre-autophagosomal | EPR22456-88 | Proximity ligation with ATG5 |
What experimental strategies identify AGP10’s post-translational modifications (PTMs)?
AGP10 undergoes phosphorylation and ubiquitination, modulating its E2 activity. Methods include:
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Immunoprecipitation (IP): Use anti-AGP10 antibodies (e.g., ARC1425) to enrich AGP10, followed by mass spectrometry to detect PTMs .
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Phospho-Specific Antibodies: Commercial kits (e.g., ab108854 ELISA) quantify phosphorylated AGP10 in serum, though cross-reactivity with other glycoproteins requires validation .
