PER26 Antibody

What is PER26 Antibody and what target does it recognize?

PER26 Antibody is a polyclonal antibody that recognizes Arabidopsis thaliana PER26 protein (UniProt Number: O22862), which is encoded by the AT2G43480 gene . This antibody is derived from rabbit hosts and is reactive against plant species, primarily Arabidopsis thaliana (Mouse-ear cress) . The immunogen used for production is a recombinant form of the PER26 protein .

PER26 belongs to the peroxidase family of proteins, which are involved in various physiological processes in plants, including responses to biotic and abiotic stresses. The antibody is designed to detect this specific peroxidase isoform in experimental systems.

What applications is PER26 Antibody validated for?

PER26 Antibody has been validated for the following applications:

The antibody components typically include:

• 200μg recombinant immunogen protein/peptide (positive control)

• 1ml pre-immune serum

• Rabbit polyclonal antibody purified by Protein A/G

This versatility allows researchers to employ the antibody in both quantitative (ELISA) and qualitative (Western blot) experimental approaches.

How should PER26 Antibody be stored and handled?

Proper storage and handling are critical for maintaining antibody performance:

• Storage temperature: -20°C or -80°C for long-term storage

• Shipping condition: Blue ice/cold chain required

• Avoid repeated freeze-thaw cycles (aliquot upon receipt)

• For working solutions, store at 4°C for up to one month

• Protect from exposure to light, particularly important for any conjugated versions

Research indicates that proper storage conditions significantly impact antibody stability and performance in experimental settings . When analyzing inconsistent results, storage conditions should be among the first variables examined.

What controls should be included when using PER26 Antibody?

Experimental design with proper controls is essential for reliable antibody-based studies:

Recommended controls:

• Positive control: Use the supplied recombinant immunogen protein/peptide

• Negative control: Pre-immune serum provided with the antibody

• Loading/normalization control: Housekeeping proteins appropriate for plant samples (e.g., actin, tubulin)

• Tissue specificity control: Samples from tissues known to express or not express PER26

• Secondary antibody-only control: To assess non-specific binding of the secondary detection system

Similar to approaches used with other antibodies in research settings , these controls help distinguish between true positive signals and experimental artifacts.

How can I optimize Western blot protocols for PER26 Antibody?

Optimization is critical for obtaining clear, specific signals:

• Sample preparation:

  • Use fresh plant tissue when possible

  • Include protease inhibitors in extraction buffer

  • Maintain cold conditions during extraction

• Blocking optimization:

  • Test both BSA and non-fat milk-based blocking solutions

  • Consider plant-specific blocking agents to reduce background

• Antibody dilution optimization:

  • Perform a dilution series (1:500, 1:1000, 1:2000) to determine optimal concentration

  • Extend primary antibody incubation to overnight at 4°C for weak signals

• Detection system selection:

  • For low abundance proteins, use high-sensitivity chemiluminescent substrates

  • Consider signal enhancement systems for difficult-to-detect targets

Similar optimization approaches have proven effective in studies utilizing plant antibodies, where tissue-specific considerations are important .

How can PER26 Antibody be integrated with NGS data analysis workflows?

Integration of antibody-based detection with next-generation sequencing provides powerful insights:

• Correlation analysis workflow:

  • Perform ChIP-seq using PER26 Antibody to identify binding regions

  • Compare binding patterns with RNA-seq expression data

  • Utilize tools like Geneious Biologics for integrated analysis

• Multi-omics approach:

  • Combine immunoprecipitation followed by mass spectrometry (IP-MS) with transcriptomics

  • Analyze resulting data with specialized NGS antibody analysis tools that can:

    • QC/trim, assemble, and merge paired-end data

    • Automatically validate sequences

    • Cluster and index annotated sequences

• Visualization strategies:

  • Use scatter plots to identify outliers and sequence distribution

  • Apply amino acid composition plots to examine variability

  • Employ heat map graphs to show relationships between genes in sequences

This integration enhances understanding of both protein presence/function and the underlying genetic regulation mechanisms.

What are the considerations for cross-reactivity assessment of PER26 Antibody?

Cross-reactivity assessment is crucial for polyclonal antibodies:

• In silico assessment:

  • Perform sequence alignment of PER26 with other peroxidase family members

  • Identify regions of high homology that may lead to cross-reactivity

• Experimental validation:

  • Test reactivity against recombinant proteins of related peroxidase family members

  • Use tissues from knockout/knockdown plants lacking PER26 expression

  • Perform peptide competition assays with specific and non-specific peptides

• Data analysis approach:

  • Quantify signal intensity ratios between target and potential cross-reactive proteins

  • Establish threshold values for specific vs. non-specific binding

Similar methodologies were employed in studies characterizing antibody specificity against related target proteins , where computational models helped disentangle binding modes associated with chemically similar ligands.

How can epitope mapping be performed to characterize PER26 Antibody binding sites?

Understanding epitope characteristics improves experimental design:

• Peptide array approach:

  • Synthesize overlapping peptides spanning the PER26 sequence

  • Probe arrays with PER26 Antibody

  • Identify peptides with strongest binding signals

• Mutation analysis:

  • Generate point mutations in recombinant PER26 protein

  • Test antibody binding to mutant proteins

  • Identify critical residues for antibody recognition

• Structural analysis:

  • If protein structure is available, map identified epitopes onto 3D structure

  • Assess epitope accessibility in native protein conformation

This approach parallels methods used in HIV antibody characterization , where crystal structures and epitope mapping revealed how antibodies recognize target proteins and viral membranes.

How should I interpret inconsistent results with PER26 Antibody?

Methodical troubleshooting approach:

• Common causes of inconsistency:

• Temporal considerations:

  • PER26 expression may vary with plant developmental stage

  • Consider diurnal variation in expression patterns

  • Document growth conditions and sampling time carefully

• Validation strategies:

  • Compare results with orthogonal detection methods (e.g., mass spectrometry)

  • Correlate protein detection with mRNA expression data

  • Consider using alternative antibodies against the same target

Similar approaches to troubleshooting were documented in antibody studies, where systematic evaluation of variables helped resolve inconsistent results .

How can I assess PER26 Antibody lot-to-lot variability?

Polyclonal antibodies can exhibit significant lot-to-lot variation:

• Quantitative assessment protocol:

  • Perform side-by-side testing of different lots

  • Generate standard curves using recombinant protein

  • Calculate detection limits, sensitivity, and signal-to-noise ratios

• Qualitative comparison:

  • Compare band patterns in Western blot

  • Assess background levels and signal specificity

  • Document differences in required working dilutions

• Record-keeping approach:

  • Maintain detailed records of lot numbers used in experiments

  • Create reference samples to test new lots against previous ones

  • Document optimization parameters for each lot

Researchers investigating long-term antibody responses have employed similar standardization approaches to ensure consistent results across multiple time points and sample sets .

How can PER26 Antibody be used in plant stress response studies?

Methodological approaches for stress response research:

• Experimental design:

  • Apply specific stressors (drought, pathogen, salt, etc.) using standardized protocols

  • Collect tissues at multiple time points after stress application

  • Include appropriate controls (unstressed, mock treatments)

• Analytical approaches:

  • Quantify changes in PER26 protein levels via Western blot densitometry

  • Assess post-translational modifications using specialized detection methods

  • Combine with cellular localization studies to track protein redistribution

• Multi-protein analysis:

  • Use co-immunoprecipitation to identify stress-specific protein interactions

  • Perform parallel analysis of related peroxidases to understand isoform-specific responses

  • Correlate protein levels with enzymatic activity assays

The dynamic nature of stress responses requires careful experimental design, similar to approaches used in tracking antibody dynamics in response to infections .

What methodologies can be employed for combining PER26 Antibody with structural biology techniques?

Integrating antibody detection with structural approaches:

• Cryo-EM applications:

  • Use PER26 Antibody for immunogold labeling

  • Visualize protein localization at ultrastructural level

  • Combine with tomography for 3D spatial information

• X-ray crystallography integration:

  • Employ antibody fragments to facilitate crystallization

  • Use antibody-facilitated crystal contacts

  • Determine antigen-antibody complex structures

• In silico structural prediction:

  • Use computational approaches to model antibody-antigen interactions

  • Predict conformational epitopes

  • Guide experimental design for structural studies

Such integrated approaches have proven valuable in understanding membrane-proximal protein interactions, as demonstrated in HIV envelope glycoprotein studies .

How can I design experiments to study post-translational modifications of PER26 using this antibody?

Methodological framework for PTM studies:

• Sample preparation strategies:

  • Include phosphatase inhibitors for phosphorylation studies

  • Use deglycosylation enzymes to assess glycosylation status

  • Employ specific extraction buffers to preserve PTMs

• Detection approaches:

  • Use Phos-tag gels to separate phosphorylated forms

  • Perform Western blots with PTM-specific detection systems

  • Use 2D gel electrophoresis to separate modified isoforms

• Validation methods:

  • Confirm PTMs with mass spectrometry

  • Use site-specific mutants to validate modification sites

  • Treat samples with specific enzymes to remove PTMs and observe mobility shifts

Similar approaches have been used in studies investigating dynamic characteristics of proteins and their modifications during stress responses .