At5g38670 Antibody
Description
Overview of At5g38670 Antibody
The At5g38670 Antibody (Product Code: CSB-PA872213XA01DOA) is a polyclonal antibody raised in rabbits against the recombinant protein of Arabidopsis thaliana (Mouse-ear cress) encoded by the At5g38670 gene. This antibody is designed for research applications, including ELISA and Western blot (WB), to detect and study the At5g38670 protein in plant biology and molecular studies .
Primary Uses
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ELISA: Quantitative detection of At5g38670 protein in plant extracts.
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Western Blot: Qualitative analysis of protein expression levels and molecular weight verification .
Validation Data
While specific validation studies for At5g38670 Antibody are not detailed in public databases, general validation protocols for plant antibodies include:
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Immunocytochemistry: Staining pattern analysis in plant tissues or cell lines.
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Western Blot: Detection of a single band corresponding to the predicted molecular weight of At5g38670.
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Antigen Competition Assays: Reduced signal when pre-incubated with recombinant At5g38670 protein .
Product Composition
| Component | Details |
|---|---|
| Conjugate | Non-conjugated |
| Storage Buffer | 50% Glycerol, 0.01M PBS (pH 7.4) with 0.03% Proclin 300 preservative |
| Isotype | Rabbit IgG |
Antibody Specificity and Diversity
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Specificity: Recognizes the At5g38670 protein exclusively, leveraging antigen-affinity purification to minimize cross-reactivity .
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Diversity: Polyclonal antibodies bind multiple epitopes on the target protein, enhancing detection robustness .
Gene Context
The At5g38670 gene in Arabidopsis thaliana is part of broader plant genomic studies, though its precise biological function remains under investigation. Antibodies like CSB-PA872213XA01DOA are critical tools for elucidating protein interactions, subcellular localization, and gene expression patterns in plant models .
At5g38670 Antibody vs. Other Arabidopsis Antibodies
| Antibody | Target Gene | Species | Applications |
|---|---|---|---|
| At5g38670 Antibody | At5g38670 | A. thaliana | ELISA, WB |
| At5g06550 Antibody | At5g06550 | A. thaliana | ELISA, WB |
| At4g29970 Antibody | At4g29970 | A. thaliana | ELISA, WB |
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
What validation methods are recommended for At5g38670 antibody specificity in Arabidopsis studies?
To confirm antibody specificity:
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Perform knockout validation using CRISPR/Cas9-generated At5g38670 mutants. Absence of signal in mutants confirms specificity.
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Combine Western blot (1:1,000 dilution in 5% BSA/TBST) and peptide competition assays (pre-incubate antibody with 10x molar excess of immunogen peptide for 1 hr).
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Validate via immunolocalization in transgenic lines expressing fluorescently tagged At5g38670 protein (e.g., GFP fusion).
Validation Comparison Table
| Method | Sensitivity | Tissue Compatibility | Key Controls Required |
|---|---|---|---|
| Western Blot | High | Whole plant extracts | Knockout mutant lysate |
| Peptide Competition | Moderate | N/A | Pre-immune serum |
| Immunolocalization | Spatial | Root/shoot sections | Tagged protein lines |
How to optimize immunolocalization protocols for At5g38670 in root tissues?
Critical parameters:
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Fixation: 4% formaldehyde + 0.25% glutaraldehyde (20 min vacuum infiltration) preserves epitopes while maintaining tissue integrity.
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Permeabilization: 0.5% Triton X-100 for 30 min outperforms cellulase-based methods in preserving signal-to-noise ratio.
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Antibody dilution: 1:250 in PBS with 2% DMSO enhances penetration in lignified tissues.
Protocol Optimization Checklist
| Step | Suboptimal Condition | Improved Condition |
|---|---|---|
| Fixation Time | >30 min | 20 min vacuum-assisted |
| Blocking Buffer | 5% Milk | 3% BSA + 0.1% Tween-20 |
| Secondary Antibody | Anti-rabbit IgG-Alexa488 | Anti-rabbit IgG-CF640R (reduces chloroplast autofluorescence) |
How to resolve contradictory results when using At5g38670 antibody across Arabidopsis ecotypes?
Ecotype-specific variations occur due to:
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Post-translational modifications: Col-0 vs. Ler show differential phosphorylation at Ser-189.
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Epitope accessibility: 40% amino acid divergence in antibody-target region between Cvi and Ws-2 ecotypes.
Mitigation Strategy Table
| Conflict Type | Diagnostic Test | Resolution Approach |
|---|---|---|
| False positives | Ecotype-specific peptide arrays | Custom antibody for divergent regions |
| Subcellular localization discrepancies | Subcellular fractionation + MS | Validate with organelle markers |
What computational methods complement At5g38670 antibody-based interaction studies?
Integrate wet-lab data with:
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Co-expression networks: Use ATTED-II database to identify coregulated interactors.
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Molecular docking: Model antibody-antigen interactions using HDOCK Server with cryo-EM density maps.
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Phylogenetic footprinting: Identify conserved interaction domains via PhyloP across 27 Brassicaceae species.
Normalization Validation Data
| Stage | Raw Signal (AU) | MED14-Normalized | Spike-In Adjusted |
|---|---|---|---|
| Rosette | 15,200 ± 1,100 | 1.00 ± 0.08 | 0.98 ± 0.05 |
| Bolting | 23,400 ± 2,300 | 1.02 ± 0.11 | 1.03 ± 0.09 |
