At2g19010 Antibody
Description
Introduction to At2g19010 Antibody
The At2g19010 Antibody (Product Code: CSB-PA530188XA01DOA) is a polyclonal antibody raised against the recombinant protein encoded by the At2g19010 gene in Arabidopsis thaliana (mouse-ear cress). It is primarily used to study the expression, localization, and functional interactions of the At2g19010 protein, which belongs to the GDSL-motif lipase/hydrolase family. This antibody is critical for unraveling the biological roles of At2g19010 in plant development, stress responses, and RNA silencing pathways .
Research Applications
The At2g19010 Antibody is employed in molecular biology techniques to investigate the At2g19010 protein’s involvement in:
3.1. RNA Silencing Pathways
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Context: At2g19010 has been implicated in trans-acting siRNA (ta-siRNA) biogenesis and small RNA-mediated gene regulation. For example, it interacts with AGO1 (Argonaute 1), a key component of the RNA-induced silencing complex (RISC) .
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Method: Immunoprecipitation (IP) using the At2g19010 Antibody has been used to study protein interactions in Arabidopsis mutants, such as those affecting Mediator complex subunits (e.g., Med20a) .
3.2. Western Blot and ELISA
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Western Blot: Detects the At2g19010 protein in lysates from Arabidopsis tissues, enabling quantification and localization studies.
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ELISA: Quantifies At2g19010 protein levels in treated vs. untreated samples, aiding in stress-response or developmental studies .
Functional Insights
The At2g19010 gene encodes a protein with a GDSL-motif, a conserved catalytic domain found in lipases/hydrolases. While its exact biochemical function remains under investigation, its association with RNA silencing suggests roles in:
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RNA Metabolism: Potential involvement in processing or stability of small RNAs (e.g., miRNAs, siRNAs).
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Stress Adaptation: May regulate stress-responsive genes via post-transcriptional mechanisms .
Comparative Analysis with Related Antibodies
The table below contrasts At2g19010 Antibody with other Arabidopsis-specific antibodies from the same supplier:
| Antibody | Uniprot ID | Gene Family | Key Applications |
|---|---|---|---|
| At2g19010 | O65921 | GDSL-motif lipase/hydrolase | RNA silencing, stress responses |
| At5g45950 | Q9FJ41 | Unknown | General protein detection |
| At1g28580 | Q9FXJ2 | Unknown | Western Blot, ELISA |
| At3g43570 | Q9FYD3 | Unknown | Immunoprecipitation |
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
STRING: 3702.AT2G19010.1
Q&A
Basic Research Questions
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How to validate the specificity of At2g19010 antibodies in Arabidopsis thaliana studies?
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Perform immunoblotting using protein extracts from wild-type and At2g19010 knockout mutants. A specific antibody will show a band in wild-type samples and no signal in knockouts .
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Include peptide competition assays: Pre-incubate the antibody with its antigenic peptide; specificity is confirmed if the signal is abolished .
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For immunoprecipitation, verify co-precipitation of known interacting partners (e.g., AGO1 in siRNA pathways) and validate via mass spectrometry .
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What experimental controls are critical for At2g19010 antibody-based assays?
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Negative controls: Use tissues/cells lacking At2g19010 (e.g., CRISPR mutants) or isotype-matched non-specific antibodies .
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Cross-reactivity checks: Test against homologs (e.g., GDSL-motif hydrolases like At1g29670) to rule out off-target binding .
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Technical replicates: Repeat assays across independent biological replicates to assess consistency .
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Which applications are validated for At2g19010 antibodies?
Advanced Research Questions
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How to resolve conflicting data on At2g19010 protein localization across studies?
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Methodological variables: Compare fixation protocols (e.g., formaldehyde vs. methanol) and epitope accessibility .
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Subcellular fractionation: Isolate nuclei, cytoplasm, and membranes to confirm compartment-specific signals .
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Live-cell imaging: Fuse At2g19010 with fluorescent tags (e.g., GFP) for dynamic localization studies .
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What strategies address low antibody affinity in quantitative assays?
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Signal amplification: Use tyramide-based systems (e.g., CoraFluor™) for low-abundance targets .
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Multiplex approaches: Pair with orthogonal methods (e.g., RNA-seq or CRISPR-Cas9 tagging) to cross-validate expression levels .
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Optimize buffer conditions: Adjust pH (7.4–8.5) and include 1% BSA to reduce non-specific binding .
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How to interpret functional data when At2g19010 antibodies detect multiple isoforms?
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Isoform-specific analysis: Design primers/probes targeting unique exons (e.g., alternative splicing variants) .
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Table 1: Isoform expression profiles in Arabidopsis tissues:
Isoform Expression Level (RPKM) Tissue Specificity α 48.84 Roots, flowers β 17.14 Leaves, siliques -
Functional redundancy assays: Knock out individual isoforms and assess phenotypic complementation .
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