At5g56900 Antibody

Basic Research Questions

• What functional roles does the At5g56900 protein play in Arabidopsis, and how can its antibody be applied to study these mechanisms?

  • Key Findings:

    • At5g56900 is a component of the autophagy conjugation pathway, critical for plant senescence and basal immunity .

    • Antibodies enable localization studies via immunofluorescence and quantify protein levels during stress responses (e.g., pathogen exposure) .

  • Methodological Guidance:

    • Use Western blotting with At5g56900 Antibody (CSB-PA801046XA01DOA, Uniprot: Q84WU9) to confirm protein expression in mutant vs. wild-type lines .

    • Pair with Arabidopsis knockout mutants to validate antibody specificity .

• How can researchers validate the specificity of At5g56900 Antibody in heterogeneous protein samples?

  • Validation Strategies:

    • Perform immunodepletion assays using recombinant At5g56900 protein to confirm signal loss .

    • Cross-validate with mass spectrometry to identify off-target binding (e.g., AMPD2 or TRIM28, common cross-reactors) .

  • Technical Specifications:

    Parameter	Detail
    Host Species	Mouse-ear cress (Arabidopsis)
    Target Epitope	Uncharacterized (Q84WU9)
    Applications	WB, IP, IF

• Which experimental models are optimal for studying At5g56900 protein-protein interactions?

  • Recommended Systems:

    • Yeast two-hybrid (Y2H) screens to map interaction partners (e.g., DDB1a in autophagy pathways) .

    • Co-IP assays in Arabidopsis protoplasts, using anti-At5g56900 for pull-downs followed by LC-MS/MS .

Advanced Research Questions

• How should researchers resolve contradictory data in co-immunoprecipitation experiments involving At5g56900?

  • Troubleshooting Framework:

    • Step 1: Verify antibody specificity via siRNA knockdown controls .

    • Step 2: Use nuclease-treated lysates to eliminate nucleic acid-mediated false interactions .

    • Step 3: Compare results across orthogonal methods (e.g., BioID proximity labeling) .

• What computational tools improve the prediction of At5g56900 antibody-antigen binding in in silico experiments?

  • Active Learning Approaches:

    • Implement Absolut! framework for predicting antibody-antigen binding landscapes, reducing required mutant testing by 35% .

    • Use AlphaFold-Multimer to model At5g56900-antibody complexes and prioritize epitope bins for mutagenesis .

• How do post-translational modifications (PTMs) of At5g56900 impact antibody performance?

  • PTM-Specific Considerations:

    • Deglycosylation assays (e.g., PNGase F treatment) to assess epitope dependence on glycosylation .

    • Phospho-specific antibodies may be required if At5g56900 activation involves phosphorylation .

Data Contradiction Analysis

Key Citations

• Antibody validation protocols:

• Functional role in autophagy:

• Computational prediction tools: