At1g69160 Antibody

What is the At1g69160 protein and what are its known functions in Arabidopsis thaliana?

At1g69160 (UniProt: Q93Z37) is a protein found in Arabidopsis thaliana, a model organism widely used in plant molecular biology research. While the specific search results don't provide extensive details on this protein's function, understanding the target is crucial for experimental design. The antibody is raised against recombinant Arabidopsis thaliana At1g69160 protein and is used primarily for detection and quantification in research applications .

The protein is studied in the context of plant molecular biology, where antibodies are essential tools for investigating protein expression, localization, and interactions. Researchers typically use such antibodies in combination with experimental techniques like ELISA and Western blotting to elucidate protein function in plant developmental processes or stress responses.

What are the validated applications for At1g69160 Antibody?

The At1g69160 Antibody has been validated for the following applications:

• ELISA (Enzyme-Linked Immunosorbent Assay): Useful for quantitative detection of the target protein in solution.

• Western Blot (WB): Enables detection of the target protein in cell or tissue lysates after separation by gel electrophoresis .

When designing experiments using this antibody, researchers should consider that it has been specifically tested and validated for these applications. While other immunological techniques might be possible, they would require additional validation by the researcher.

What are the optimal storage and handling conditions for At1g69160 Antibody?

Proper storage and handling are critical for maintaining antibody functionality:

• Storage temperature: Upon receipt, store at -20°C or -80°C to maintain activity

• Avoid repeated freeze-thaw cycles: These can damage antibody structure and reduce binding efficiency

• Storage buffer: The antibody is supplied in a buffer containing:

  • 50% Glycerol

  • 0.01M PBS (pH 7.4)

  • 0.03% Proclin 300 as preservative

For routine use, aliquoting the antibody into single-use volumes before freezing can prevent repeated freeze-thaw cycles. Always handle the antibody according to good laboratory practices, including wearing gloves to prevent contamination.

How should researchers validate the specificity of At1g69160 Antibody results?

Antibody validation is a critical step in ensuring experimental reliability. For At1g69160 Antibody, researchers should:

• Include proper controls:

  • Positive control: Sample known to express At1g69160

  • Negative control: Sample known not to express the target

  • Secondary antibody-only control: To assess background binding

• Perform blocking experiments: Pre-incubate the antibody with purified recombinant At1g69160 protein before the primary detection experiment to demonstrate specificity.

• Confirm results with alternative methods: Use techniques like RT-PCR to verify that protein detection correlates with gene expression patterns.

This validation approach follows general principles used in antibody-based research, as demonstrated in studies using other antibodies where specificity validation was crucial for reliable results .

What are typical working dilutions for At1g69160 Antibody in different applications?

While the specific optimal dilutions for this particular antibody are not explicitly stated in the search results, researchers should consider the following general guidelines based on similar antibodies:

These are starting points, and researchers should perform dilution series experiments to determine the optimal concentration for their specific samples and experimental conditions. The antibody is antigen-affinity purified, which generally allows for higher dilutions compared to non-purified antibodies .

How can researchers address potential cross-reactivity when using At1g69160 Antibody?

Cross-reactivity is a significant concern in antibody-based research. For At1g69160 Antibody:

• Species specificity: The antibody is specifically raised against Arabidopsis thaliana protein . When using it with other plant species, researchers should:

  • Perform sequence alignment analysis between Arabidopsis At1g69160 and homologous proteins in the target species

  • Include appropriate controls from Arabidopsis as reference points

  • Validate binding specificity in the new species through additional experiments

• Addressing non-specific binding:

  • Optimize blocking conditions using different blocking agents (BSA, normal serum, or commercial blockers)

  • Increase the number and duration of washing steps

  • Test different dilutions of primary and secondary antibodies to minimize background

Similar approaches have been used in other antibody research to address cross-reactivity issues, as demonstrated in immunological studies where specificity was crucial for interpreting results .

What are effective strategies for troubleshooting weak or absent signals when using At1g69160 Antibody?

When encountering weak or absent signals with At1g69160 Antibody, researchers should systematically investigate:

• Protein extraction and sample preparation issues:

  • Ensure complete extraction using buffers appropriate for plant tissues

  • Consider the presence of interfering compounds in plant samples that may require specific extraction methods

  • Verify protein integrity through total protein staining

• Antibody-specific factors:

  • Check antibody viability (avoid expired antibodies or those subjected to multiple freeze-thaw cycles)

  • Reduce antibody dilution to increase signal strength

  • Extend primary antibody incubation time or temperature

• Detection system optimization:

  • Use enhanced chemiluminescence (ECL) substrates with higher sensitivity

  • Increase exposure time when developing Western blots

  • Consider signal amplification methods like biotin-streptavidin systems

This methodical approach to troubleshooting is consistent with best practices in antibody-based research, where optimization of multiple parameters is often necessary to achieve reliable results .

How can At1g69160 Antibody be incorporated into multi-parameter experimental designs?

Advanced research often requires simultaneous analysis of multiple proteins. For incorporating At1g69160 Antibody into such designs:

• Multiplex immunofluorescence strategies:

  • Combine At1g69160 Antibody with antibodies against other proteins expressed in distinct subcellular compartments

  • Use secondary antibodies conjugated to different fluorophores

  • Implement appropriate controls to assess spectral overlap and antibody cross-reactivity

• Sequential immunoblotting approaches:

  • Strip and reprobe membranes to detect multiple proteins on the same blot

  • Use differently sized target proteins that can be clearly distinguished

  • Consider using antibodies raised in different host species to enable simultaneous detection

• Co-immunoprecipitation experiments:

  • Use At1g69160 Antibody to pull down protein complexes

  • Analyze interaction partners through mass spectrometry or Western blotting

These approaches are consistent with advanced immunological research methods that maximize information obtained from limited biological samples, similar to strategies employed in other antibody-based studies .

What modifications to standard protocols are necessary when using At1g69160 Antibody in tissues with high levels of interfering compounds?

Plant tissues often contain compounds that can interfere with antibody-based detection:

• Addressing phenolic compounds and polysaccharides:

  • Include polyvinylpyrrolidone (PVP) or polyvinylpolypyrrolidone (PVPP) in extraction buffers to absorb phenolics

  • Add higher concentrations of reducing agents (β-mercaptoethanol or DTT) to prevent oxidation

  • Consider extraction with TCA/acetone to precipitate proteins while removing interfering compounds

• Protocol modifications for difficult tissues:

  • Extend blocking times to reduce non-specific binding

  • Add detergents (Tween-20, Triton X-100) to washing buffers to reduce background

  • Include protease inhibitors in all buffers to prevent degradation of target proteins

• Sample preparation considerations:

  • For tissues with high lipid content, include additional delipidation steps

  • Consider subcellular fractionation to enrich for compartments containing the target protein

These methodological adjustments are particularly important in plant research, where tissue-specific interfering compounds can significantly impact antibody performance and experimental outcomes.

How can computational approaches enhance the interpretation of At1g69160 Antibody experimental data?

Modern research increasingly integrates computational methods with experimental data:

• Quantitative image analysis for immunolocalization:

  • Use software tools to quantify signal intensity and co-localization with other markers

  • Apply deconvolution algorithms to improve resolution in confocal microscopy

  • Implement machine learning approaches for unbiased pattern recognition

• Integrating antibody data with -omics datasets:

  • Compare protein detection patterns with transcriptomic data

  • Correlate protein abundance with metabolomic profiles

  • Place findings in the context of known protein interaction networks

• Modeling antibody binding dynamics:

  • Apply mathematical models similar to those described for other antibody systems to understand binding kinetics

  • Predict potential cross-reactivity based on structural modeling

  • Estimate detection limits through simulation of antibody-antigen interactions

This integration of computational approaches with experimental data represents the frontier of antibody-based research, enabling more comprehensive interpretation of results and generation of testable hypotheses for future experiments.