At3g48460 Antibody
Description
Gene and Protein Background
The AT3G48460 gene in Arabidopsis thaliana encodes a protein classified within the GDSL esterase/lipase family. Key attributes include:
| Attribute | Description |
|---|---|
| Gene Symbol | AT3G48460 |
| Entrez Gene ID | 824005 |
| Protein Name | GDSL-like Lipase/Acylhydrolase superfamily protein |
| Organism | Arabidopsis thaliana (thale cress) |
| Function | Involved in fatty acid metabolism, embryo development, and stress responses |
Orthologs include Os03g0844600 in Oryza sativa (rice), sharing functional similarities in lipid-related pathways .
Antibody Development and Production
The AT3G48460 antibody was developed to study the protein’s localization and function. Key steps include:
Antibody Preparation Protocol
| Step | Method |
|---|---|
| Antigen Source | Recombinant SFAR4 protein (signal peptide excluded) expressed in E. coli |
| Purification | HisTrap affinity chromatography |
| Immunization | Rabbit polyclonal antibody production (70-day protocol) |
| Validation | SDS-PAGE, Western blot, and tissue-specific GUS staining in transgenic plants |
This antibody was used to confirm SFAR4 expression in embryo protrusions and its role in fatty acid reduction .
Role in Fatty Acid Metabolism
The AT3G48460 protein (SFAR4) is implicated in:
-
Fatty acid reduction during seed maturation.
-
Embryo development, with intense expression in embryo protrusions .
-
Stress responses, though specific mechanisms remain under investigation.
Experimental Validation
| Assay | Outcome |
|---|---|
| GUS Staining | Localized SFAR4 expression in embryo protrusions across 10+ transgenic lines |
| Genomic PCR | T-DNA insertion validation in mutant plants |
| Southern Blot | Confirming single-copy T-DNA integration |
These experiments established SFAR4’s spatial and temporal expression patterns .
Comparison with Other GDSL Esterases
AT3G48460 belongs to a family of enzymes with diverse catalytic roles. Below is a functional comparison:
| Enzyme | Catalytic Activity | Biological Role |
|---|---|---|
| AT3G48460 (SFAR4) | Fatty acid reduction | Seed maturation, embryo development |
| Other GDSL esterases | Lipid hydrolysis, acyl transfer | Pathogen defense, cell wall remodeling |
While structural homology exists, substrate specificity and biological roles diverge .
Challenges and Future Directions
-
Limited Commercial Availability: Current antibodies are primarily for academic use, with no reported therapeutic applications.
-
Functional Complexity: The GDSL family’s diverse enzymatic activities necessitate targeted substrate profiling.
-
Ortholog Studies: Comparative analysis with rice (Os03g0844600) could reveal conserved mechanisms in lipid metabolism .
Table 1: Gene Orthologs
| Organism | Gene ID | Protein ID |
|---|---|---|
| Arabidopsis thaliana | AT3G48460 | NP_190416.1 |
| Oryza sativa | Os03g0844600 | NP_001051872.1 |
Table 2: Antibody Validation Techniques
| Technique | Purpose | Outcome |
|---|---|---|
| SDS-PAGE | Confirm protein size and purity | ~30 kDa band detected |
| Western Blot | Validate antibody specificity | Single band in SFAR4-expressing tissues |
| GUS Staining | Map tissue-specific expression | Signal in embryo protrusions |
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
