TPS27 Antibody

What is the TPS27 antibody and what does it target?

TPS27 antibody is a rabbit polyclonal antibody that specifically recognizes the TPS27 protein in plants, particularly Arabidopsis thaliana. This antibody targets the terpene synthase-like sequence-1, 8-cineole (TPS-CIN), which is involved in plant secondary metabolism . The antibody is generated using recombinant Arabidopsis thaliana TPS27 protein as the immunogen, ensuring specificity for plant research applications .

What experimental applications is the TPS27 antibody validated for?

The TPS27 antibody has been validated for several research applications including:

• Enzyme Immunoassay (EIA)

• General Immunoassay protocols

• Enzyme-Linked Immunosorbent Assay (ELISA)

• Western Blot analysis

This versatility makes it suitable for both protein quantification and qualitative detection experiments in plant biology research.

What is the recommended storage protocol for TPS27 antibody?

For optimal antibody performance and longevity, the TPS27 antibody should be stored at -20°C or -80°C upon receipt. It's crucial to avoid repeated freeze-thaw cycles as these can degrade antibody quality and reduce binding efficiency . For researchers planning extensive experimental series, aliquoting the antibody upon receipt is recommended to minimize freeze-thaw cycles.

How should I determine the optimal dilution of TPS27 antibody for my specific experimental system?

Determining optimal antibody dilution requires systematic titration experiments:

• Begin with a broad dilution range (e.g., 1:500, 1:1000, 1:2000, 1:5000) in your application of choice

• Run parallel experiments with positive controls (known TPS27-expressing samples) and negative controls

• Evaluate signal-to-noise ratio at each dilution

• Select the dilution that provides maximum specific signal with minimal background

• Fine-tune with narrower dilution ranges around the identified optimum

Remember that optimal dilutions may vary between applications – a dilution that works for Western blot may not be optimal for ELISA.

What are the recommended blocking agents when using TPS27 antibody in immunoblotting?

While specific blocking optimization data for TPS27 antibody is not detailed in the available literature, plant antibody research typically benefits from:

• 5% non-fat dry milk in TBS-T (standard starting point)

• 3-5% BSA in TBS-T (alternate for phospho-specific targets)

• Commercial plant-optimized blocking buffers (for samples with high background)

Test multiple blocking agents in parallel when establishing your protocol, as the optimal agent may depend on your specific plant tissue and extraction method.

How can I validate the specificity of the TPS27 antibody in my experimental system?

Comprehensive antibody validation should include:

• Positive controls: Use recombinant Arabidopsis thaliana TPS27 protein or known TPS27-expressing tissue

• Negative controls: Include samples from:

  • TPS27 knockout mutants

  • Non-plant tissues

  • Pre-immune serum controls

• Peptide competition assay: Pre-incubate the antibody with excess immunogenic peptide to demonstrate signal reduction

• Molecular weight verification: Confirm that detected bands match the expected molecular weight of TPS27

• Multiple detection methods: Validate across different applications (e.g., if positive in Western blot, confirm with ELISA)

What are the most effective protein extraction protocols for detecting TPS27 in plant tissues?

Effective TPS27 protein extraction requires preserving protein integrity while maximizing yield:

• Extraction buffer composition:

  • 50 mM Tris-HCl (pH 7.5)

  • 150 mM NaCl

  • 1% Triton X-100 (or alternative non-ionic detergent)

  • 0.5% sodium deoxycholate

  • Protease inhibitor cocktail optimized for plants

• Critical extraction steps:

  • Maintain cold conditions throughout (4°C)

  • Use mechanical disruption (e.g., bead-beating) for complete tissue homogenization

  • Centrifuge at high speed (≥14,000×g) to remove debris

  • Consider ultrafiltration for sample concentration if TPS27 expression is low

• Tissue-specific considerations:

  • Young tissues typically yield better results than senescent tissues

  • Consider developmental timing as terpene synthase expression varies throughout plant development

What strategies can address weak or absent signal when using TPS27 antibody in Western blots?

If experiencing weak or no signal when using TPS27 antibody:

• Sample preparation issues:

  • Increase protein concentration

  • Verify protein transfer efficiency with reversible stains

  • Try gentler extraction methods to preserve epitope integrity

• Detection optimization:

  • Increase antibody concentration (use less dilute solution)

  • Extend primary antibody incubation (overnight at 4°C)

  • Switch to more sensitive detection systems (ECL Plus vs. standard ECL)

  • Try longer exposure times

• Expression factors:

  • Verify TPS27 expression in your specific tissue/conditions (check transcriptomic data)

  • Consider induction treatments (e.g., jasmonic acid) to upregulate terpene synthesis

  • Harvest tissue at developmental stages known for higher TPS27 expression

How can I minimize cross-reactivity with other terpene synthases when using TPS27 antibody?

The TPS gene family has many members with structural similarities. To improve specificity:

• Stringent washing conditions:

  • Increase salt concentration in wash buffers (up to 500 mM NaCl)

  • Add low concentrations of SDS (0.1%) to wash buffers

  • Extend washing duration and increase washing steps

• Antibody pre-absorption:

  • Incubate antibody with protein extracts from tissues expressing other TPS family members but not TPS27

  • Filter this solution and use the pre-absorbed antibody for your experiments

• Alternative detection approaches:

  • Consider epitope-tagged recombinant TPS27 expression for unambiguous detection

  • Combine with mass spectrometry for definitive identification

How does the performance of polyclonal TPS27 antibody compare to monoclonal alternatives?

While specific comparative data for TPS27 antibodies is not detailed in the search results, general principles apply:

Select based on your experimental priorities: polyclonals offer broader epitope recognition while monoclonals provide consistent reproducibility.

What are the key considerations when using TPS27 antibody across different plant species?

When extending TPS27 antibody use beyond Arabidopsis:

• Sequence homology assessment:

  • Perform sequence alignment of TPS27 between Arabidopsis and your target species

  • 70% identity in immunogenic regions suggests potential cross-reactivity

• Validation experiments:

  • Always include Arabidopsis samples as positive controls

  • Run Western blots with both species to compare band patterns

  • Consider epitope mapping if working extensively with non-Arabidopsis species

• Dilution and protocol adaptation:

  • Start with more concentrated antibody solutions (2-3× more concentrated)

  • Optimize incubation conditions specifically for the new species

  • Consider species-specific protein extraction protocols to preserve epitopes

How can TPS27 antibody be effectively used in studies of plant defense responses?

TPS27 (TPS-CIN) is involved in terpene production relevant to plant defense. Research applications include:

• Stress response studies:

  • Track TPS27 protein levels following biotic stressors (herbivory, pathogen infection)

  • Compare TPS27 protein abundance across resistant vs. susceptible plant varieties

  • Correlate protein levels with volatile terpene emissions

• Spatio-temporal analysis:

  • Use immunohistochemistry to localize TPS27 in different tissues

  • Couple with GC-MS analysis of monoterpenes to correlate protein levels with enzymatic output

  • Study developmental regulation of defense compound production

• Signal transduction investigation:

  • Monitor TPS27 protein levels following application of defense hormones (jasmonic acid, salicylic acid)

  • Compare with transcriptional analysis to identify post-transcriptional regulation

What protocols enable quantitative analysis of TPS27 protein abundance in plant tissues?

For quantitative measurement of TPS27:

• Quantitative ELISA approaches:

  • Develop a standard curve using recombinant TPS27 protein

  • Ensure extraction buffers are compatible with ELISA protocols

  • Include internal standards to normalize between experimental batches

• Quantitative Western blot:

  • Use housekeeping proteins (e.g., actin, tubulin) as loading controls

  • Employ fluorescent secondary antibodies for wider linear range

  • Perform densitometry analysis with appropriate software

  • Include dilution series of standards for calibration

• Data normalization considerations:

  • Account for total protein content (Bradford/BCA assay)

  • Consider tissue-specific reference proteins

  • Report relative abundance changes rather than absolute values unless validated standards are used

How might TPS27 antibody be utilized in emerging plant biotechnology applications?

Emerging applications for TPS27 antibody include:

• Metabolic engineering verification:

  • Confirm protein expression in transgenic plants with modified terpene biosynthetic pathways

  • Monitor protein stability in plants engineered for enhanced essential oil production

  • Validate subcellular targeting in compartmentalized biosynthesis approaches

• Protein-protein interaction studies:

  • Use for co-immunoprecipitation to identify interaction partners

  • Apply in proximity ligation assays to visualize protein complexes in situ

  • Combine with mass spectrometry for interactome analysis

• Climate change response research:

  • Monitor changes in TPS27 protein levels under altered CO₂ and temperature conditions

  • Correlate with volatile emission profiles to understand adaptive responses

  • Develop early biochemical markers for plant stress under changing climate conditions

What opportunities exist for integrating TPS27 protein detection with multi-omics approaches?

Integration opportunities include:

• Proteomics-metabolomics integration:

  • Correlate TPS27 protein abundance with terpene metabolite profiles

  • Identify post-translational modifications affecting enzyme activity

  • Develop predictive models linking protein levels to metabolic outputs

• Transcriptome-proteome correlation:

  • Compare TPS27 transcript levels with protein abundance to identify regulatory mechanisms

  • Identify conditions with post-transcriptional regulation

  • Develop correction factors for transcript-based predictions of enzyme activity

• Systems biology approaches:

  • Position TPS27 within broader signaling and metabolic networks

  • Identify rate-limiting steps in terpene biosynthesis

  • Model flux through biosynthetic pathways based on enzyme abundance

What quality control metrics should be monitored when using TPS27 antibody in long-term research programs?

For consistent results across extended research projects:

• Antibody performance tracking:

  • Maintain a positive control sample as reference standard

  • Document signal intensity and background levels for each experiment

  • Record lot numbers and create control charts to track performance over time

• Storage and handling validation:

  • Test aliquots after various storage durations to establish stability profile

  • Compare fresh vs. stored antibody performance periodically

  • Document freeze-thaw stability with performance metrics

• Regular specificity confirmation:

  • Periodically revalidate with knockout/negative controls

  • Compare detection patterns across different tissues/conditions

  • Consider epitope mapping if inconsistencies arise

What are the critical parameters to report when publishing research using TPS27 antibody?

When publishing TPS27 antibody-based research, report:

• Antibody details:

  • Full commercial information (manufacturer, catalog number: orb786349)

  • Lot number (for reproducibility)

  • Host species (rabbit)

  • Clonality (polyclonal)

  • Immunogen details (recombinant Arabidopsis thaliana TPS27 protein)

• Experimental conditions:

  • Complete protocol details (dilutions, incubation times/temperatures)

  • Blocking reagents and concentrations

  • Detection method specifications

  • Image acquisition parameters

• Validation evidence:

  • Controls used (positive, negative, loading)

  • How specificity was confirmed

  • Representative images of full blots/results

  • Quantification method details