At5g03820 Antibody
Description
Basic Information
The At5g03820 antibody is cataloged as a commercial research product with the following specifications:
| Parameter | Details |
|---|---|
| Product Name | At5g03820 Antibody |
| Product Code | CSB-PA878645XA01DOA |
| Target Protein | At5g03820 (UniProt ID: Q9LZC5) |
| Host Species | Arabidopsis thaliana (Mouse-ear cress) |
| Available Formats | 2 ml or 0.1 ml aliquots |
| Applications | Western blotting, immunohistochemistry, ELISA |
This antibody is part of a broader catalog of antibodies targeting Arabidopsis proteins, enabling researchers to study plant-specific molecular pathways .
Molecular Characterization
The At5g03820 gene encodes a protein whose exact biological function remains under investigation. Key features include:
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Protein Structure: Predicted molecular weight and domain architecture (derived from UniProt data).
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Expression Profile: Likely expressed in specific tissues or under certain stress conditions, based on homology to related plant proteins .
Limitations: No peer-reviewed studies directly characterizing the At5g03820 antibody’s specificity or performance were identified in the provided sources. Structural details (e.g., epitope mapping) are unavailable .
Research Applications
While direct studies on this antibody are scarce, its potential applications align with broader uses of plant-specific antibodies :
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Protein Localization: Tracking spatial expression of At5g03820 in plant tissues.
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Functional Genomics: Knockout/knockdown studies to elucidate gene function.
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Stress Response Studies: Investigating roles in abiotic/biotic stress pathways.
Challenges and Future Directions
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Specificity Validation: Independent verification of cross-reactivity and batch consistency is needed.
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Functional Data: Linking At5g03820 protein activity to phenotypic traits in Arabidopsis.
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
STRING: 3702.AT5G03820.1
Q&A
How can I validate the specificity of the At5g03820 antibody in Western blot assays?
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Method: Perform immunoblotting using protein extracts from Arabidopsis thaliana wild-type (Col-0) and at5g03820 knockout mutants. A specific antibody will show a band in wild-type samples and no signal in mutants.
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Controls: Include a recombinant At5g03820 protein as a positive control and pre-immune serum as a negative control.
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Troubleshooting: If nonspecific bands appear, optimize blocking conditions (e.g., 5% nonfat milk in TBST) or try alternative electrophoresis buffers (e.g., Tris-Glycine vs. Bis-Tris).
What experimental designs are optimal for studying At5g03820 protein localization in root tissues?
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Approach:
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Immunolocalization: Fix roots in 4% paraformaldehyde, embed in paraffin, and use the antibody at 1:200 dilution with Alexa Fluor 488-conjugated secondary antibodies.
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Confocal imaging: Combine with propidium iodide counterstaining to visualize cell walls.
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Validation: Compare localization patterns with GFP-tagged At5g03820 transgenic lines.
How to resolve contradictions in At5g03820 expression data between qRT-PCR and Western blot analyses?
| Factor | qRT-PCR | Western Blot |
|---|---|---|
| Sensitivity | Detects mRNA levels (femtomolar) | Detects protein (picogram range) |
| Post-transcriptional regulation | No | Affected by translation efficiency |
| Antibody specificity | N/A | Risk of cross-reactivity |
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Solution: Perform time-course experiments to account for delays in transcription-translation and use protease inhibitors during protein extraction.
What advanced techniques can characterize At5g03820’s interaction partners?
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Co-Immunoprecipitation (Co-IP):
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Prepare native protein extracts from inflorescence tissue.
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Use the At5g03820 antibody conjugated to Protein A/G beads.
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Identify interactors via mass spectrometry.
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Bimolecular Fluorescence Complementation (BiFC): Co-express At5g03820-YFPn and candidate interactors-YFPc in protoplasts.
How does pH variability affect At5g03820 antibody performance in ELISA?
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Optimal conditions:
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Coating buffer: Carbonate-bicarbonate (pH 9.6) enhances antigen adsorption.
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Washing buffer: PBS (pH 7.4) with 0.05% Tween-20 reduces nonspecific binding.
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Critical step: Pre-adsorb the antibody with pH-adjusted plant extract to neutralize cross-reactive epitopes.
What statistical methods are robust for quantifying At5g03820 expression gradients in leaf primordia?
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Spatial analysis: Use Fiji/ImageJ with the “Radial Profile” plugin to measure fluorescence intensity gradients.
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Normalization: Express data as a ratio to internal controls (e.g., actin or tubulin signal in the same section).
How to differentiate between isoform-specific and cross-reactive signals in At5g03820 immunostaining?
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Strategy:
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Isoform-selective knockdown: Use CRISPR-Cas9 to delete individual exons unique to each isoform.
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Epitope mapping: Synthesize overlapping 15-mer peptides spanning the At5g03820 sequence to identify antibody-binding regions.
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What controls are essential when using the At5g03820 antibody in chromatin immunoprecipitation (ChIP)?
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Mandatory controls:
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Input DNA (no immunoprecipitation)
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IgG isotype control
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At5g03820 knockout mutant chromatin
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Validation: Perform qPCR on known target loci (e.g., promoters of genes regulated by At5g03820).
How to optimize transient expression assays for testing At5g03820 antibody specificity in Nicotiana benthamiana?
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Infiltrate leaves with Agrobacterium strains expressing:
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At5g03820-GFP
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Endogenous N. benthamiana homologs (e.g., NbSGT1)
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Compare antibody signals between treatments to assess cross-reactivity.
What metrics indicate successful At5g03820 antibody validation for publication?
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Key criteria:
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Signal absence in knockout mutants (≥3 biological replicates)
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Correlation (R² ≥ 0.85) between antibody signal and transcript abundance across tissues
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Consistent performance in ≥2 independent assays (e.g., Western blot and immunohistochemistry).
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