At5g10900 Antibody
Description
Molecular Target: AT5G10900 Protein
The At5g10900 gene encodes a calcineurin-like metallo-phosphoesterase superfamily protein (UniProt ID: Q9LEV0) . This protein, designated PP7L (Ser/Thr-protein phosphatase 7 inactive homolog), plays critical roles in:
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Chloroplast development and membrane structure maintenance .
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Autophagic processes, including ciliogenesis via OFD1 and IFT20 regulation .
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Lymphocyte survival and proliferation in animals (by homology) .
PP7L lacks catalytic phosphatase activity but participates in protein-protein interactions, influencing signaling pathways .
3.1. Protein Localization and Expression Analysis
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Western Blot: Detects PP7L at ~32 kDa in Arabidopsis lysates, with validation in knockout lines (e.g., pp7l-1 mutants) confirming specificity .
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Immunohistochemistry (IHC): Used to visualize PP7L distribution in plant tissues, such as chloroplast-rich cells in leaves .
3.2. Functional Studies
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Chloroplast Development: PP7L knockout mutants (pp7l-En-1) exhibit impaired chloroplast morphology and reduced photosynthetic efficiency .
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Stress Response: PP7L interacts with autophagy-related proteins under oxidative stress, suggesting a role in cellular repair .
4.1. Western Blot Performance
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Lane Specificity: Bands observed at 32 kDa in wild-type Arabidopsis lysates (e.g., HeLa, HT1080 cells), absent in ATG5 knockout lines .
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Cross-Reactivity: No off-target binding reported in human or mouse samples .
4.2. Immunohistochemistry
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Tissue Staining: Strong signal in paraffin-embedded Arabidopsis root and stem sections, correlating with PP7L’s role in membrane structure maintenance .
Future Directions
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Mechanistic Studies: Elucidate PP7L’s role in chloroplast-mitochondrial crosstalk.
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Agricultural Biotechnology: Engineer PP7L-overexpressing crops for enhanced stress tolerance.
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
Based on the analysis of current research, here is a structured FAQ addressing key scientific considerations for ATG5 antibodies (note: At5g10900 refers to an Arabidopsis thaliana gene unrelated to human ATG5; we address the well-characterized human ATG5 antibody research from available data):
How to validate ATG5 antibody specificity in autophagy studies?
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Methodological approach:
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Perform triple validation using:
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Critical data interpretation:
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What experimental designs effectively link ATG5 expression to apoptotic pathways?
How to resolve conflicting data on ATG5's role in viral replication?
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Multiparametric approach:
What advanced techniques map conformational epitopes of anti-ATG5 antibodies?
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Integrated structural biology workflow:
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Hydrogen-deuterium exchange mapping
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Cryo-EM of antibody-antigen complexes
How to improve signal-to-noise ratio in ATG5 immunohistochemistry?
How to distinguish autophagy-dependent vs independent ATG5 functions?
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Three-axis confirmation system:
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Genetic: CRISPR ATG5 KO + rescue experiments
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Pharmacological: 3-MA vs. wortmannin treatment controls
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Temporal analysis: Early (0-6 hr) vs late-phase (12-24 hr) effects
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Key references: (ATG5 functional domains), (antibody internalization dynamics), (neutralization mechanisms)
