At1g51650 Antibody

What is the AT1G51650 antibody and what does it target?

The AT1G51650 antibody specifically targets the ATP synthase subunit epsilon, mitochondrial protein in plants. This protein is a component of the mitochondrial ATP synthase complex, which plays a crucial role in energy production through oxidative phosphorylation . The antibody is commonly used in plant molecular biology research to study mitochondrial function, energy metabolism, and related cellular processes. When implementing this antibody in your research, remember that it recognizes the epsilon subunit specifically, which allows for targeted analysis of this component within the larger ATP synthase complex.

Which plant species can be studied using the AT1G51650 antibody?

The AT1G51650 antibody demonstrates cross-reactivity with multiple plant species, making it versatile for comparative studies. Based on specificity and cross-reaction data, the antibody can be effectively used with the following plant species:

The diverse cross-reactivity profile makes this antibody particularly valuable for comparative studies across different plant families . When designing experiments involving multiple species, preliminary validation through Western blotting is recommended to confirm recognition efficiency across your specific samples.

What are the standard storage and handling conditions for the AT1G51650 antibody?

For optimal research results, proper storage and handling of the AT1G51650 antibody are critical. The antibody is typically supplied in lyophilized form and should be handled according to these guidelines:

• Store the lyophilized antibody in a manual defrost freezer

• Avoid repeated freeze-thaw cycles as they may compromise antibody functionality

• The product is shipped at 4°C, and upon receipt, it should be immediately stored at the recommended temperature

• When reconstituting the lyophilized antibody, use sterile techniques and store aliquots to minimize freeze-thaw cycles

Following these storage protocols is essential for maintaining antibody stability and ensuring reproducible experimental results across extended research projects.

Which experimental techniques are compatible with the AT1G51650 antibody?

The AT1G51650 antibody can be employed in multiple experimental techniques commonly used in plant molecular biology research. Based on antibody characteristics and research applications, the following techniques are compatible:

• Western Blotting: The primary application for detecting ATP synthase epsilon subunit in protein extracts

• Immunohistochemistry (IHC): For visualizing the protein's localization in plant tissue sections

• Immunoprecipitation (IP): To isolate the target protein and its interacting partners

• ELISA: For quantitative detection of the target protein

• Immunofluorescence: For subcellular localization studies in fixed cells or tissues

When implementing any of these techniques, optimization of antibody dilution is essential. Start with the manufacturer's recommended dilution and adjust based on your specific experimental conditions and sample types. For complex plant tissues with high polyphenol content, additional blocking steps may be necessary to reduce background signal.

How should samples be prepared for optimal results with the AT1G51650 antibody?

Sample preparation significantly impacts the success of experiments using the AT1G51650 antibody. For optimal results with plant samples, follow these methodological guidelines:

• For protein extraction, use buffers containing protease inhibitors to prevent degradation of the target protein

• Include reducing agents like DTT or β-mercaptoethanol in sample buffers for Western blotting

• For mitochondrial enrichment protocols (recommended for higher sensitivity):

  • Homogenize plant tissue in isolation buffer (0.3M sucrose, 50mM HEPES, 2mM EDTA, pH 7.5)

  • Perform differential centrifugation to isolate the mitochondrial fraction

  • Verify mitochondrial enrichment using known markers

• For fixed tissue preparations, optimize fixation time to maintain epitope accessibility while preserving tissue architecture

These methodological considerations help ensure that the epitope recognized by the antibody remains intact and accessible throughout the experimental procedure, leading to more reliable and reproducible results.

How can the AT1G51650 antibody be used to study mitochondrial dysfunction in stress responses?

The ATP synthase epsilon subunit plays a crucial role in energy production, making the AT1G51650 antibody valuable for investigating mitochondrial responses to environmental stresses. Advanced research applications include:

• Quantitative analysis of ATP synthase subunit expression changes under various stress conditions

• Correlation between ATP synthase complex integrity and mitochondrial function during stress

• Comparative analysis across stress-tolerant and stress-sensitive plant varieties

• Investigation of post-translational modifications of the epsilon subunit under stress conditions

For effective stress response studies, establish time-course experiments with careful controls. Western blot quantification should be normalized to stable mitochondrial markers rather than whole-cell housekeeping proteins. Combine antibody-based detection with functional assays (oxygen consumption, ATP production) to correlate protein expression with mitochondrial performance.

What approaches can be used to validate AT1G51650 antibody specificity in new experimental systems?

When introducing the AT1G51650 antibody to a new plant species or experimental system, rigorous validation is essential. Advanced researchers should employ these methodological approaches:

• Perform Western blot analysis with:

  • Positive controls from validated species (e.g., Arabidopsis thaliana)

  • Negative controls lacking the target protein (if available, knockout/knockdown lines)

  • Peptide competition assays to confirm specific binding

• Compare immunolocalization patterns with established mitochondrial markers

• Validate target detection through mass spectrometry analysis of immunoprecipitated proteins

• Compare reactivity across tissue types and developmental stages to establish baseline patterns

These validation steps are particularly important when working with plant species not listed in the cross-reactivity data or when investigating tissues with complex protein compositions.

How can the AT1G51650 antibody contribute to research on mitochondrial biogenesis and assembly?

The ATP synthase epsilon subunit is essential for proper assembly and function of the ATP synthase complex. Advanced research using this antibody can investigate:

• Temporal sequence of ATP synthase complex assembly during plant development

• Coordination between nuclear and mitochondrial genomes in encoding ATP synthase components

• Impact of mutations in related subunits on epsilon subunit incorporation

• Effects of retrograde signaling on ATP synthase assembly under various conditions

For assembly studies, combine the AT1G51650 antibody with antibodies against other ATP synthase subunits and utilize techniques such as Blue Native-PAGE followed by Western blotting to analyze intact complexes. This approach allows for the assessment of complex assembly states rather than just individual subunit expression.

How should researchers address potential cross-reactivity concerns with the AT1G51650 antibody?

Despite its specificity, all antibodies carry potential cross-reactivity risks that should be systematically addressed:

• Perform side-by-side comparisons across different plant species using consistent protein loading

• Include appropriate molecular weight markers to confirm target band identity

• For novel applications, consider these additional validation steps:

  • Pre-absorb the antibody with purified target protein to eliminate specific binding

  • Use genetic approaches (RNAi, CRISPR) to create samples with reduced target expression

  • Compare results with alternative antibodies targeting the same protein, if available

When publishing, report all validation steps performed and discuss any unexpected bands or staining patterns to maintain transparency about potential cross-reactivity issues.

What are the common pitfalls in quantitative analysis using the AT1G51650 antibody?

Quantitative analysis of Western blots or immunohistochemistry using the AT1G51650 antibody requires awareness of several methodological constraints:

To overcome these challenges, researchers should:

• Establish a standard curve with known concentrations of recombinant protein

• Use multiple technical and biological replicates

• Employ appropriate statistical analyses for semi-quantitative Western blot data

• Consider complementary approaches like mass spectrometry for absolute quantification

What strategies can resolve weak or absent signals when using the AT1G51650 antibody?

When signal strength is insufficient despite confirmed presence of the target protein, systematic troubleshooting is necessary:

• Antibody concentration: Optimize by testing a range of dilutions, typically starting with 1:500 to 1:2000 for Western blotting

• Sample preparation modifications:

  • Increase protein loading (up to 50μg for total protein extracts)

  • Use mitochondrial enrichment to concentrate the target protein

  • Optimize extraction buffer components to improve protein solubility

• Detection system enhancements:

  • Switch to more sensitive detection systems (e.g., from colorimetric to chemiluminescent)

  • Increase exposure time (while monitoring background)

  • Consider signal amplification systems like biotin-streptavidin

For each modification, include appropriate controls to ensure that any improvements in signal are specific to the target protein rather than increased background.

How can researchers differentiate between specific and non-specific binding when using the AT1G51650 antibody?

Distinguishing specific from non-specific binding is crucial for accurate data interpretation. Advanced researchers should employ these analytical approaches:

• Compare observed band patterns with theoretical molecular weights for the target protein

• Perform peptide competition assays by pre-incubating the antibody with excess target peptide

• Include samples with different expression levels of the target protein (natural variation or engineered)

• Analyze samples from related species with known sequence differences in the target protein

When non-specific binding is detected, optimize blocking conditions (type of blocking agent, concentration, and incubation time) and washing stringency (salt concentration and detergent levels in wash buffers).

How can the AT1G51650 antibody be incorporated into multi-omics experimental designs?

The AT1G51650 antibody can serve as a valuable component in integrated multi-omics studies of plant mitochondrial function:

• Proteomics integration:

  • Use the antibody for immunoprecipitation followed by mass spectrometry to identify interaction partners

  • Compare antibody-based quantification with mass spectrometry-based protein abundance measurements

• Transcriptomics correlation:

  • Analyze correlation between transcript levels of AT1G51650 and protein abundance under various conditions

• Metabolomics connections:

  • Correlate ATP synthase epsilon subunit levels with ATP/ADP ratios and TCA cycle metabolites

• Phenomics applications:

  • Connect molecular-level changes detected by the antibody to plant-level phenotypic responses

When designing such integrated studies, ensure consistent sample handling across different analytical platforms and develop robust statistical frameworks for correlating data across omics layers.

What considerations are important when adapting AT1G51650 antibody protocols for new plant species?

When extending AT1G51650 antibody applications to plant species beyond those listed in cross-reactivity data, researchers should:

• Perform sequence homology analysis of the target protein across species to predict likelihood of antibody recognition

• Optimize protein extraction protocols specific to the new species, accounting for differences in:

  • Cell wall composition

  • Secondary metabolite profiles

  • Protein-to-non-protein ratio in tissues

• Test multiple antibody concentrations over a wider range than typically used

• Consider epitope retrieval techniques if standard protocols yield poor results

• Validate results using orthogonal approaches (e.g., mass spectrometry identification of the target protein)

This systematic approach to protocol adaptation increases the likelihood of successful antibody application in new species while maintaining scientific rigor.