At2g19893 Antibody

What is At2g19893 Antibody and what are its key specifications?

At2g19893 Antibody (product code CSB-PA647277XA01DOA) is a polyclonal antibody raised in rabbits against recombinant Arabidopsis thaliana At2g19893 protein. It is supplied in liquid form with a storage buffer containing 50% Glycerol, 0.01M PBS (pH 7.4), and 0.03% Proclin 300 as a preservative. The antibody is antigen-affinity purified and designed specifically for research applications .

Key specifications include:

• Type: Polyclonal Antibody

• Host: Rabbit

• Target: At2g19893 protein from Arabidopsis thaliana

• Applications: ELISA, Western Blot (WB)

• Storage: -20°C or -80°C (avoid repeated freeze-thaw cycles)

• Format: Non-conjugated, liquid

• Purification: Antigen Affinity Purified

What are the validated applications for At2g19893 Antibody?

At2g19893 Antibody has been validated specifically for Enzyme-Linked Immunosorbent Assay (ELISA) and Western Blot (WB) applications for the identification of the target antigen. These techniques allow researchers to detect, quantify, and characterize the At2g19893 protein in various experimental contexts .

It is essential to note that this antibody is strictly for research use only and not intended for diagnostic or therapeutic procedures, as explicitly stated by the manufacturer .

What is the expected lead time and proper storage protocol for At2g19893 Antibody?

The At2g19893 Antibody is made-to-order with an expected lead time of 14-16 weeks, which researchers should factor into project planning timelines . Upon receipt, the antibody should be stored at either -20°C or -80°C, with care taken to avoid repeated freeze-thaw cycles as this can compromise antibody activity and specificity .

How should researchers validate At2g19893 Antibody specificity for their experiments?

Comprehensive validation is critical for ensuring reliable results with At2g19893 Antibody. Researchers should implement a multi-step validation strategy:

• Positive and negative controls: Include samples known to express (positive) or not express (negative) the At2g19893 protein.

• Blocking peptide competition: Pre-incubate the antibody with the immunizing peptide to confirm signal specificity.

• Knockout/knockdown verification: Test the antibody in samples where the At2g19893 gene has been silenced or deleted.

• Cross-reactivity assessment: Examine potential cross-reactivity with related proteins.

• Multiple detection methods: Confirm results using different techniques when possible.

This validation approach aligns with the broader scientific consensus that proper antibody validation is essential for research reproducibility, regardless of whether antibodies are animal-derived or non-animal-derived .

What considerations should researchers make when designing Western Blot experiments using At2g19893 Antibody?

For optimal Western Blot results with At2g19893 Antibody, consider these methodological parameters:

These parameters should be systematically optimized for your specific experimental conditions .

How can researchers effectively design experiments to study post-translational modifications using At2g19893 Antibody?

When investigating post-translational modifications (PTMs) of the At2g19893 protein, researchers should implement specialized approaches:

• Preservation of modification status: Include appropriate inhibitors in lysis buffers (phosphatase inhibitors for phosphorylation studies, deubiquitinase inhibitors for ubiquitination studies).

• Enrichment strategies: For low-abundance modified forms, implement immunoprecipitation or affinity purification before detection.

• Controls for modification specificity: Include samples treated with enzymes that remove the modification of interest.

• Epitope accessibility consideration: Be aware that PTMs might affect antibody binding, potentially leading to false-negative results.

• Orthogonal validation: Confirm findings with mass spectrometry or other methods that directly detect the modification .

How can At2g19893 Antibody be utilized in studying protein-protein interactions in Arabidopsis thaliana?

At2g19893 Antibody can serve as a powerful tool for investigating protein-protein interactions through multiple complementary approaches:

• Co-immunoprecipitation (Co-IP): Use the antibody to pull down At2g19893 protein complexes, followed by mass spectrometry to identify interaction partners.

• Proximity ligation assay (PLA): Combine At2g19893 Antibody with antibodies against suspected interaction partners to visualize interactions in situ with single-molecule sensitivity.

• Immunofluorescence co-localization: Use the antibody alongside antibodies against potential interacting proteins to determine subcellular co-localization patterns.

• Protein complex isolation: Employ the antibody in native protein complex isolation followed by biochemical characterization.

• Validation of screen-identified interactions: Confirm interactions identified in yeast two-hybrid or other screening approaches .

What approaches can researchers use to improve reproducibility when working with At2g19893 Antibody?

The reproducibility of experiments using At2g19893 Antibody can be enhanced through several methodological considerations:

• Standardized protocols: Develop and adhere to detailed protocols for sample preparation, antibody handling, and assay conditions.

• Antibody validation documentation: Maintain comprehensive records of antibody validation experiments and results.

• Lot-to-lot consistency checks: Verify performance when using new antibody lots.

• Positive and negative controls: Include appropriate controls in every experiment.

• Multiple detection methods: Confirm key findings using orthogonal techniques.

• Quantitative analysis: Implement standardized quantification methods with appropriate statistical analysis.

• Detailed method reporting: Document all experimental parameters in publications, including antibody catalog number, dilution, incubation conditions, and validation approach .

How can researchers effectively use At2g19893 Antibody in comparative studies across different Arabidopsis ecotypes or related species?

For comparative studies across Arabidopsis ecotypes or related plant species, researchers should consider:

• Epitope conservation analysis: Perform bioinformatic analysis to determine conservation of the epitope region across the species of interest.

• Pilot validation: Test antibody reactivity with samples from each ecotype or species before proceeding with full experiments.

• Standardized sample preparation: Develop consistent protein extraction methods across different plant materials.

• Normalization strategies: Implement robust normalization methods using conserved proteins as loading controls.

• Complementary approaches: Supplement antibody-based detection with transcript analysis to correlate protein and mRNA levels.

• Cross-validation: When possible, confirm key findings with alternative methods .

What are common causes of false negative results with At2g19893 Antibody and their solutions?

False negative results when using At2g19893 Antibody may stem from several sources:

These troubleshooting approaches can help overcome technical challenges and improve detection sensitivity .

How can researchers optimize immunohistochemistry protocols for At2g19893 Antibody in plant tissues?

Although immunohistochemistry (IHC) is not listed among the validated applications for At2g19893 Antibody, researchers might explore this application with appropriate optimization:

• Fixation optimization: Test different fixatives (e.g., paraformaldehyde, glutaraldehyde) and fixation times to balance antigen preservation with tissue morphology.

• Antigen retrieval: Evaluate heat-induced or enzymatic antigen retrieval methods to expose epitopes masked during fixation.

• Permeabilization: Optimize detergent concentration and treatment time to improve antibody penetration.

• Background reduction: Implement strategies such as endogenous peroxidase quenching and appropriate blocking to minimize non-specific signals.

• Detection system selection: Choose a detection system with appropriate sensitivity for the expected abundance of At2g19893.

It's worth noting that synthetic antibodies historically have not worked well for IHC methods, with only a limited number of non-animal-derived antibodies demonstrating adequate performance in IHC applications .

What approaches can be used to quantify At2g19893 protein levels accurately using this antibody?

For accurate quantification of At2g19893 protein levels:

• Standard curve generation: Where possible, use recombinant At2g19893 protein at known concentrations to create a reference standard.

• Loading controls: Implement appropriate loading controls that remain stable across experimental conditions.

• Multiple technical replicates: Perform at least three technical replicates to account for assay variability.

• Linear range determination: Ensure measurements fall within the linear range of detection by testing serial dilutions.

• Digital imaging quantification: Use appropriate software for densitometry analysis with background subtraction.

• ELISA quantification: Consider developing a quantitative ELISA using the antibody for more precise measurements.

• Statistical validation: Apply appropriate statistical tests to validate the significance of observed differences .

How might new antibody generation approaches improve research on At2g19893 protein complexes?

Recent innovations in antibody technology offer potential advantages for studying At2g19893 protein complexes:

• Fusion protein approach: Creating fusion proteins of At2g19893 with its interacting partners could stabilize complexes during antibody generation, potentially yielding antibodies that specifically recognize the complex rather than individual proteins.

• Complex-specific epitopes: This approach may generate antibodies that specifically recognize interface epitopes only present when proteins interact.

• In-cell measurements: The resulting antibodies could enable direct measurement of protein complexes on live cells, similar to the BTLA-HVEM complex research that demonstrated "direct measurement on live cells using a complex-specific monoclonal antibody" .

• Disease relevance: Such approaches could facilitate studies of At2g19893's potential role in plant stress responses or disease resistance mechanisms .

What are the comparative advantages and limitations of using animal-derived versus non-animal-derived antibody approaches for At2g19893 research?

When considering antibody selection for At2g19893 research, it's important to understand the tradeoffs between animal-derived (like this antibody) and non-animal-derived alternatives:

This comparison highlights that both approaches have complementary strengths, and the choice should depend on specific research requirements .

How can researchers integrate At2g19893 Antibody data with other omics approaches for systems-level understanding in Arabidopsis research?

To integrate At2g19893 Antibody data with other omics approaches for comprehensive insights:

• Multi-omics correlation: Analyze correlations between protein levels (detected with At2g19893 Antibody), transcriptomics data (RNA-seq), and potentially metabolomics data related to At2g19893 function.

• Network analysis: Use protein interaction data generated with At2g19893 Antibody as input for network analysis tools to identify functional modules.

• Temporal studies: Combine antibody-based detection with time-series experiments to determine dynamic changes in protein levels and modifications.

• Spatial integration: Correlate protein localization data with cell-type-specific transcriptomics to understand tissue-specific roles.

• Perturbation analysis: Use At2g19893 Antibody to measure protein responses across various genetic backgrounds or environmental conditions, integrating with transcriptomic changes.

• Functional validation: Use knowledge gained from omics integration to design targeted validation experiments using the antibody for specific hypotheses .