The AT1G28650 gene encodes a protein with the following attributes:
Attribute
Details
Gene Symbol
AT1G28650
Entrez Gene ID
839765
Full Name
GDSL-like Lipase/Acylhydrolase superfamily protein
Synonyms
F1K23.27, F1K23_27
Organism
Arabidopsis thaliana (thale cress)
Protein Function
Putative involvement in lipid metabolism and enzymatic acyltransferase activities
The GDSL-like Lipase/Acylhydrolase family is conserved in plants and plays roles in processes such as lipid remodeling, pathogen defense, and stress adaptation.
Key Challenges in Plant Antibody Production
Low Success Rates with Peptide Immunogens:
Peptide-based antibodies often fail to detect targets in situ due to poor epitope accessibility or improper folding.
Example: Only 55% of 70 recombinant protein antibodies showed detectable signals after affinity purification.
Signal Peptide Optimization:
Heavy-chain signal peptides (SPs) with higher essential amino acid (EAA) content improve transient antibody production in HEK293 cells.
Light-chain SPs (e.g., Vκ1) exhibit better homogeneity and lower EAA counts, enhancing yield.
Affinity Purification:
Crude antisera often require purification via Protein A/G or recombinant protein affinity columns to reduce non-specific binding.
Potential Applications and Research Gaps
While no direct studies on the AT1G28650 antibody exist, its utility can be inferred from analogous antibodies in plant biology:
Application
Relevance to AT1G28650
Protein Localization
Determining subcellular localization (e.g., cytoplasm, membrane) via immunofluorescence.
Functional Studies
Investigating roles in lipid metabolism, stress responses, or enzymatic activity.
Biomarker Discovery
Identifying disease-related or stress-induced protein modifications.
Research Gaps:
No published studies validate the AT1G28650 antibody’s specificity or efficacy.
Limited data on cross-reactivity with homologous GDSL-like proteins in other species.
Comparative Analysis of Antibody Production Strategies
Below is a comparison of methods used in plant antibody development, relevant to AT1G28650:
Strategy
Advantages
Limitations
Recombinant Protein Immunization
Higher success rates vs. peptide immunogens.
Requires soluble, folded recombinant protein.
Peptide Immunization
Lower cost; easier epitope design.
High failure rates; poor in situ detection.
Signal Peptide Optimization
Improves transient production yields.
Requires extensive screening of SP variants.
Future Directions
To advance AT1G28650 antibody research:
Epitope Mapping: Define linear or conformational epitopes for improved specificity.
Validation in Mutant Backgrounds: Confirm antibody specificity using at1g28650 knockout lines.
