LTPG31 Antibody

What is LTPG31 and why is it significant in plant biology research?

LTPG31 (encoded by At5g64080 in Arabidopsis thaliana) belongs to a family of GPI-anchored lipid transfer proteins (LTPgs). This protein is particularly significant because it is one of the few LTPg genes that show upregulation in syncytia, as opposed to the majority of LTPg genes which are suppressed . LTPG proteins are involved in cuticular lipid transport, with the LTPG family comprising 34 members in Arabidopsis distributed across all five chromosomes .

LTPG31 research provides insights into:

• Plant lipid transport mechanisms

• Cell wall formation and integrity

• Plant responses to environmental stresses

• Cellular trafficking of hydrophobic compounds

LTPG31 antibodies enable researchers to track the protein's localization, expression patterns, and functional roles in plant development and stress responses.

How do LTPG antibody epitopes differ from other plant cell wall antibodies?

The epitope recognition of LTPG antibodies, including those for LTPG31, differs significantly from other plant cell wall antibodies:

• Protein vs. carbohydrate epitopes: While many plant cell wall antibodies (like CCRC-M26) recognize carbohydrate structures such as arabinogalactans , LTPG antibodies recognize protein epitopes within the lipid transfer protein domains.

• Conserved cysteine motifs: LTPG antibodies typically recognize regions containing the characteristic cysteine residues that form disulfide bridges. These eight cysteine residues are highly conserved among LTPg proteins and create a specific structural signature .

• Domain specificity: Effective LTPG31 antibodies should target unique regions that differentiate it from other LTPG family members, particularly focusing on:

  • The non-conserved regions outside the LTP domain

  • The unique C-terminal region before the GPI anchor attachment site (ω-site)

  • Specific sequences in motifs 1, 2, or 3 that show variation between LTPG family members

Unlike antibodies against arabinogalactans that may require 30+ residues for optimal binding , LTPG antibodies typically recognize smaller protein epitopes but must be carefully designed to avoid cross-reactivity with other LTPG family members.

What are the recommended validation methods for LTPG31 antibodies?

Rigorous validation of LTPG31 antibodies should include multiple complementary approaches:

Primary validation methods:

• Western blotting with negative controls: Compare wild-type plants with ltpg31 knockout/knockdown mutants to confirm antibody specificity

• Immunoprecipitation followed by mass spectrometry: Verify that the antibody specifically pulls down LTPG31 rather than other LTPG family members

• Peptide competition assay: Pre-incubate the antibody with the immunizing peptide to confirm epitope specificity

Secondary validation methods:

• Immunolocalization in transgenic plants: Compare antibody staining patterns with fluorescently tagged LTPG31 fusion proteins (such as LTPG31-GFP)

• Tissue-specific expression analysis: Validate that immunostaining matches known tissue-specific expression patterns of LTPG31

• Cross-reactivity assessment: Test against recombinant proteins of closely related LTPG family members

Documentation requirements:

• Antibody metadata including: clone name, species immunized, isotype, epitope details

• Images of all validation experiments with appropriate controls

• Detailed protocols specifying critical parameters (antibody dilutions, incubation times, etc.)

Following these validation protocols ensures that experimental findings can be confidently attributed to LTPG31 rather than to cross-reactivity with other LTPG family members.

What are the optimal conditions for immunolocalization of LTPG31 in plant tissues?

Successful immunolocalization of LTPG31 requires careful sample preparation and optimization:

Fixation protocol:

• Recommended fixative: 4% paraformaldehyde in PBS (pH 7.4) supplemented with 0.1% Triton X-100

• Fixation time: 2-4 hours at room temperature or overnight at 4°C

• Critical consideration: Overfixation can mask the epitope, while underfixation leads to poor structural preservation

Antigen retrieval options:

• Enzymatic approach: Mild treatment with cellulase (0.1%) and pectinase (0.05%) for 10-15 minutes can improve antibody accessibility

• Heat-mediated approach: Citrate buffer (pH 6.0) at 80°C for 10 minutes

• Note: Test both methods to determine optimal signal-to-noise ratio for LTPG31

Blocking and antibody incubation parameters:

• Blocking agent: 3% BSA with 0.1% Tween-20 in PBS for 1 hour

• Primary antibody dilution: Start at 1:500 and titrate (typically 1:200-1:1000)

• Incubation time: Overnight at 4°C for primary antibody

• Secondary antibody: Use fluorescently labeled secondaries matching the primary antibody species (typically at 1:1000 dilution)

Special considerations for GPI-anchored proteins:

• Detergent concentration is critical; too high will extract GPI-anchored proteins

• Consider membrane preservation techniques if studying membrane localization

• Compare with a fluorescently tagged LTPG31 as a reference for localization pattern

The procedure should be validated by comparing results with the pattern observed in plants expressing LTPG31-YFP fusion proteins under native promoter control, similar to the approach used for other LTPG family members .

How should researchers design co-immunoprecipitation experiments with LTPG31 antibodies?

Co-immunoprecipitation (Co-IP) with LTPG31 antibodies requires specific considerations for GPI-anchored membrane proteins:

Sample preparation protocol:

• Tissue selection: Use tissues with known LTPG31 expression (e.g., epidermal tissues)

• Buffer composition:

  • Base buffer: 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA

  • Detergent options:

    • Mild: 0.5-1% NP-40 or 1% Triton X-100 (preserves most interactions)

    • Stringent: 0.1% SDS + 1% Triton X-100 (reduces weak/non-specific interactions)

  • Protease inhibitor cocktail (PMSF, leupeptin, aprotinin)

  • Phosphatase inhibitors if studying phosphorylation-dependent interactions

Co-IP procedure optimization:

• Pre-clearing: Incubate lysate with protein A/G beads (1-2 hours at 4°C) before antibody addition

• Antibody binding: Use 2-5 μg antibody per 500 μg protein lysate

• Incubation time: Overnight at 4°C with gentle rotation

• Washing stringency: Perform 4-5 washes with decreasing detergent concentration

• Elution options:

  • Denaturing: SDS sample buffer at 95°C (disrupts all interactions)

  • Non-denaturing: Excess peptide competition (preserves complex integrity)

Control experiments (essential):

• Negative control: IgG from same species as LTPG31 antibody

• Knockout/knockdown validation: Compare results with ltpg31 mutant plant tissues

• Reciprocal Co-IP: Confirm interactions by IP with antibodies against interacting partners

• Input control: Reserve 5-10% of pre-IP lysate to confirm starting protein levels

Detection methods:

• Western blot: For known/suspected interacting partners

• Mass spectrometry: For unbiased identification of co-precipitated proteins

This approach has proven effective for studying protein interactions of GPI-anchored proteins in plants, similar to methods used for characterizing interactions of other LTPG family members .

What methods are most effective for quantifying LTPG31 protein expression levels?

Accurate quantification of LTPG31 protein expression requires techniques optimized for membrane-associated proteins:

Western blot quantification:

• Extraction protocol optimization:

  • Buffer: 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1% Triton X-100, 0.1% SDS

  • Include 6M urea for enhanced GPI-anchored protein solubilization

  • Fresh protease inhibitor cocktail

• Sample processing:

  • Heat samples at 65°C instead of 95°C to prevent aggregation

  • Load equal total protein amounts (15-30 μg) determined by Bradford assay

• Quantification approach:

  • Use infrared fluorescent secondary antibodies for wider linear range

  • Include recombinant LTPG31 protein standards for absolute quantification

  • Normalize to multiple reference proteins (e.g., H3, actin, GAPDH)

ELISA-based quantification:

• Develop sandwich ELISA using two antibodies recognizing different LTPG31 epitopes

• Generate standard curve using recombinant LTPG31 protein

• Process samples with specialized membrane protein extraction buffers

Comparative analysis table:

Statistical considerations:

• Minimum of 3 biological replicates

• Use ANOVA with post-hoc tests for multiple condition comparisons

• Consider non-parametric tests if normality assumptions aren't met

The approach selected should mirror techniques that have been successful for quantifying other LTP family proteins in plants , with modifications specific to the biochemical properties of LTPG31.

How can researchers effectively use antibodies to study LTPG31 interactions with other cell wall and membrane components?

Investigating LTPG31 interactions requires specialized approaches that account for its membrane association and potential lipid cargo:

Proximity-based interaction methods:

• In situ proximity ligation assay (PLA):

  • Requires antibodies against LTPG31 and suspected interaction partner from different species

  • Can visualize interactions in fixed tissues with subcellular resolution

  • Quantification possible by counting fluorescent dots per cell

• Cross-linking approaches:

  • Chemical cross-linkers (DSP, formaldehyde) to stabilize transient interactions

  • UV cross-linking to capture direct protein-protein contacts

  • Must optimize cross-linker concentration to avoid non-specific aggregation

Biochemical fractionation strategies:

• Detergent-resistant membrane (DRM) isolation:

  • Extract with 1% Triton X-100 at 4°C

  • Separate by sucrose gradient ultracentrifugation

  • Analyze LTPG31 distribution among fractions via western blot

  • Compare with known lipid raft markers (e.g., flotillin)

• Cell wall protein fractionation:

  • Sequential extraction with CaCl₂, EGTA, and LiCl

  • Compare LTPG31 extraction pattern with known cell wall proteins

  • Test effect of phospholipase C treatment to release GPI-anchored proteins

Advanced microscopy approaches:

• FRET/FLIM analysis:

  • Generate fluorescently tagged LTPG31 and potential interactors

  • Measure energy transfer efficiency as indicator of proximity

  • Control for protein expression levels to avoid overexpression artifacts

• Super-resolution microscopy:

  • STORM or PALM techniques can resolve co-localization below diffraction limit

  • Requires special fluorophore-conjugated antibodies or tags

This multi-technique approach has proven successful for characterizing the interactions of GPI-anchored proteins like LTPG in Arabidopsis and can be specifically adapted for LTPG31.

What are the considerations for using LTPG31 antibodies in studying abiotic stress responses in plants?

LTPG31 and related proteins play important roles in plant responses to environmental stresses, requiring specific experimental approaches:

Stress treatment design principles:

• Stress selection and relevance:

  • Heavy metal stress shows strong effects on LTPg expression

  • Drought stress affects cuticle composition where LTPgs function

  • Control treatment severity through time-course and dose-response experiments

• Tissue sampling considerations:

  • Focus on epidermal tissues where LTPG31 is expressed

  • Sample at multiple timepoints (early, intermediate, late responses)

  • Include recovery phase samples to assess reversibility

Antibody-based analytical approaches:

• Time-course immunoblotting:

  • Track LTPG31 protein levels during stress and recovery phases

  • Compare with transcript levels (RT-qPCR) to identify post-transcriptional regulation

  • Assess protein modifications (phosphorylation, glycosylation) using specialized stains

• Comparative immunolocalization:

  • Map subcellular redistribution of LTPG31 under stress conditions

  • Co-stain with organelle markers to track trafficking changes

  • Quantify changes in plasma membrane vs. intracellular localization

Specialized methodological adaptations:

• Protein stability assessment:

  • Cycloheximide chase experiments to determine if stress affects LTPG31 turnover

  • Proteasome inhibitor (MG132) treatments to assess degradation pathways

• Post-translational modification analysis:

  • Phosphorylation: Immunoprecipitate LTPG31 followed by phospho-specific staining

  • Lipidation: Analyze GPI-anchor integrity under stress conditions

  • Conformation changes: Limited proteolysis assays before/after stress

Data integration framework:

• Correlate protein-level changes with physiological parameters

• Compare wild-type responses with ltpg31 mutants to establish causality

• Consider redundancy with other LTPg family members in stress responses

This research framework builds on established approaches for studying stress-responsive proteins in Arabidopsis while accounting for the specific properties of LTPG31.

How can antibodies help differentiate between the diverse biological functions of LTPG family members?

The LTPG family comprises 34 members in Arabidopsis with potentially overlapping but distinct functions. Antibody-based approaches can help delineate their specific roles:

Comparative expression mapping:

• Multi-antibody tissue profiling:

  • Generate antibodies against select LTPG family members (focusing on LTPG5, LTPG31)

  • Perform immunohistochemistry on the same tissue sections

  • Create expression maps highlighting zones of unique vs. overlapping expression

  • Quantify relative expression levels in different tissues and developmental stages

• Developmental timeline analysis:

  • Track expression patterns through key developmental transitions

  • Correlate with developmental events (e.g., cuticle formation, flowering)

  • Compare with RNA-seq data to identify post-transcriptional regulation

Functional differentiation approaches:

• Immunoprecipitation followed by lipid binding analysis:

  • IP different LTPG proteins using specific antibodies

  • Analyze bound lipids by mass spectrometry

  • Compare lipid binding profiles to identify substrate preferences

  • Create a matrix of LTPG-lipid interactions to map functional specialization

• Antibody-based protein complex comparison:

  • IP different LTPG proteins and identify interacting partners

  • Construct protein interaction networks for each LTPG

  • Identify unique vs. shared interaction partners

  • Correlate with known biological pathways

Technical specifications for differential antibody development:

• Epitope selection strategies:

  • Target variable regions outside conserved cysteine motifs

  • Focus on C-terminal regions before GPI attachment sites

  • Design peptide antigens from unique sequence stretches

• Cross-reactivity control measures:

  • Test against recombinant proteins of all closely related LTPGs

  • Validate in knockout/knockdown mutants for target LTPG

  • Pre-absorb antibodies with recombinant proteins of related family members

This systematic approach has been used to differentiate functions within other protein families in plants and can be applied to elucidate the specific biological roles of LTPG31 versus other family members .

How should researchers address potential cross-reactivity between LTPG31 antibodies and other LTPG family members?

Cross-reactivity is a major concern when working with members of the LTPG family due to their sequence and structural similarities:

Comprehensive cross-reactivity assessment protocol:

• In silico analysis:

  • Perform multiple sequence alignment of all 34 LTPG family members

  • Calculate sequence identity percentages between LTPG31 and other LTPGs

  • Identify regions of highest similarity, particularly in motifs 1, 2, and 3

  • Predict potential cross-reactive epitopes using epitope prediction algorithms

• Experimental validation:

  • Express recombinant fragments of closely related LTPG proteins

  • Perform dot blot or western blot with serial dilutions of each protein

  • Create a cross-reactivity matrix showing reactivity percentages

  • Determine minimum detectable concentration for each family member

Cross-reactivity mitigation strategies:

• Antibody purification approaches:

  • Affinity purification against LTPG31-specific peptide

  • Negative selection against closely related LTPG family members

  • Test pre-absorption with recombinant proteins of closely related LTPGs

• Experimental design controls:

  • Include ltpg31 knockout/knockdown tissues as negative controls

  • Use tissues with known LTPG expression profiles based on transcriptomics

  • Perform parallel experiments with fluorescent protein fusions (LTPG31-GFP)

Decision matrix for interpreting potential cross-reactivity:

The rigorous validation approach is critical since the LTPG family shows conservation in key structural motifs while differing in specific functions, as demonstrated in previous studies of LTPG proteins in Arabidopsis .

What are common technical challenges when working with LTPG31 antibodies and how can they be overcome?

Researchers often encounter several technical challenges when working with antibodies against GPI-anchored proteins like LTPG31:

Challenge 1: Low signal intensity in immunodetection

• Potential causes:

  • Low natural abundance of LTPG31

  • Epitope masking due to protein conformation or interactions

  • GPI-anchor affecting antibody accessibility

• Solutions:

  • Sample enrichment: Concentrate membrane fractions before analysis

  • Modified fixation: Test alternative fixatives or reduced fixation times

  • Epitope retrieval: Apply gentle detergent treatment or heat-mediated antigen retrieval

  • Signal amplification: Use tyramide signal amplification or enhanced chemiluminescence

Challenge 2: High background in immunolocalization

• Potential causes:

  • Non-specific binding to other plant cell wall components

  • Autofluorescence from phenolic compounds or chlorophyll

  • Secondary antibody cross-reactivity with endogenous plant proteins

• Solutions:

  • Blocking optimization: Test different blockers (BSA, milk, normal serum)

  • Autofluorescence reduction: Use Sudan Black B (0.1%) treatment

  • Additional washing: Increase wash times and add 0.05-0.1% Tween-20

  • Antibody dilution: Titrate antibody to find optimal signal-to-noise ratio

  • Fluorophore selection: Choose fluorophores outside plant autofluorescence spectrum

Challenge 3: Inconsistent immunoprecipitation results

• Potential causes:

  • GPI-anchor affecting protein solubility

  • Weak antibody-antigen affinity

  • Detergent disrupting critical epitopes

  • Transient or context-dependent protein interactions

• Solutions:

  • Buffer optimization: Test different detergent types and concentrations

  • Cross-linking: Use reversible cross-linkers to stabilize complexes

  • Antibody immobilization: Covalently couple antibody to beads

  • Proximity labeling: Consider BioID or APEX2 approaches as alternatives

Challenge 4: Conflicting results between antibody detection and fluorescent fusion proteins

• Potential causes:

  • Fluorescent tag affecting protein localization or function

  • Antibody cross-reactivity with related proteins

  • Overexpression artifacts with fusion proteins

• Solutions:

  • Validation triangulation: Compare with transcript data (RNA-seq, in situ hybridization)

  • Complementary approaches: Use both N- and C-terminal tagged versions

  • Expression control: Use native promoters instead of strong constitutive promoters

  • Genetic validation: Test antibody specificity in knockout/knockdown lines

These troubleshooting approaches are based on successful strategies used with other plant membrane proteins and modified for the specific challenges presented by LTPG family proteins .

How can researchers interpret apparent discrepancies between LTPG31 protein levels detected by antibodies and transcript data from qPCR or RNA-seq?

Discrepancies between protein and transcript levels are common in biological systems and require careful interpretation, especially for regulatory proteins like LTPG31:

Systematic analysis framework:

• Validation of both data types:

  • Confirm antibody specificity through knockout controls

  • Verify qPCR primer specificity through melt curves and sequencing

  • Check RNA-seq data alignment uniqueness to LTPG31 vs. other family members

  • Determine technical variability through replicate analysis

• Time-course considerations:

  • Protein expression typically lags behind transcript changes

  • Compare different time points to identify temporal relationships

  • Consider protein half-life vs. transcript degradation rates

  • Analyze both steady-state and dynamic response conditions

Common discrepancy patterns and interpretations:

Mechanistic investigation approaches:

• Post-transcriptional regulation:

  • Polysome profiling to assess translation efficiency

  • RNA-binding protein immunoprecipitation to identify regulators

  • miRNA analysis to identify potential inhibitory mechanisms

• Post-translational regulation:

  • Ubiquitination analysis through IP and western blot

  • Proteasome and autophagy inhibitors to assess degradation pathways

  • Pulse-chase experiments to determine protein half-life

• Technical reconciliation approaches:

  • Absolute quantification of both transcript (copies/cell) and protein (molecules/cell)

  • Normalization to appropriate reference genes/proteins for each method

  • Mathematical modeling to account for temporal dynamics

This framework has been applied successfully to understand post-transcriptional regulation in plant systems, including cases where transcript and protein levels do not correlate, as seen in studies of stress responses affecting LTPg family members and mechanisms affecting other GPI-anchored proteins .

How might new antibody technologies enhance research on LTPG31 and related proteins?

Emerging antibody technologies offer significant potential to advance LTPG31 research beyond current capabilities:

Advanced detection technologies:

• Single-domain antibodies (nanobodies):

  • Smaller size (15 kDa vs. ~150 kDa) enables better penetration into dense plant tissues

  • Can access epitopes in tight spaces, including cell wall-membrane interfaces

  • Potential applications:

    • Super-resolution microscopy of LTPG31 in membrane microdomains

    • In vivo imaging with membrane-permeable nanobodies

    • Improved immunoprecipitation of intact membrane complexes

• Proximity-dependent labeling antibodies:

  • Antibodies conjugated to enzymes like APEX2, BioID, or TurboID

  • Label proteins within 10-50 nm radius of the target

  • Applications for LTPG31 research:

    • Mapping the proximal proteome around LTPG31 in native conditions

    • Identifying transient interaction partners during stress responses

    • Defining membrane microdomain composition around LTPG31

Emerging antibody production methods relevant to LTPG research:

• Phage display libraries without animal immunization:

  • Similar to approaches used for cell wall antibodies

  • Allows selection of antibodies against conserved plant epitopes

  • Can generate antibodies against multiple family members simultaneously

  • Enables epitope fine-tuning through directed evolution

• AI-designed synthetic antibodies:

  • Computational design of antibodies targeting specific LTPG31 epitopes

  • Optimization for minimal cross-reactivity with other LTPG family members

  • Structure-based antibody engineering for improved affinity and specificity

Transformative experimental approaches:

• Intrabodies for in vivo functional studies:

  • Express antibody fragments inside plant cells

  • Target specific domains to inhibit function

  • Applications:

    • Block LTPG31 lipid binding without genetic modification

    • Disrupt specific protein-protein interactions

    • Create rapid conditional knockdowns

• Stimuli-responsive antibody systems:

  • Light-activatable antibody fragments

  • Chemical-inducible binding systems

  • Enable:

    • Temporal control of LTPG31 inhibition

    • Cell-type specific functional analysis

    • Reversible perturbation of LTPG31 function

These technologies build upon established antibody approaches while offering new capabilities particularly valuable for studying membrane-anchored proteins like LTPG31, where traditional approaches face significant limitations .

What interdisciplinary approaches combining LTPG31 antibodies with other techniques could advance understanding of plant lipid transport?

Integrative methodologies combining antibody-based techniques with complementary approaches can provide comprehensive insights into LTPG31 function:

Structural biology integration:

• Antibody-assisted cryo-EM studies:

  • Use antibody fragments (Fab, scFv) to stabilize LTPG31 for structural determination

  • Map lipid-binding pocket architecture in different functional states

  • Visualize LTPG31 in complex with interaction partners

  • Resolution potential: 2.5-4Å for membrane protein complexes

• Hydrogen-deuterium exchange mass spectrometry with antibody footprinting:

  • Map conformational changes upon lipid binding

  • Identify regions protected by antibody binding

  • Characterize dynamic structural transitions during lipid loading/unloading

Advanced imaging combinations:

• Correlative light and electron microscopy (CLEM):

  • Immunofluorescence to locate LTPG31 in tissue sections

  • Follow with EM imaging of the same section for ultrastructural context

  • Visualize LTPG31 at the cell wall-plasma membrane interface

  • Track vesicular trafficking pathways of LTPG31

• Expansion microscopy with immunolabeling:

  • Physical expansion of specimens to achieve super-resolution

  • Combine with LTPG31 antibody labeling

  • Resolve spatial relationships with cell wall components

  • Map nanoscale distribution patterns in membrane microdomains

Multi-omics integration frameworks:

• Antibody-based isolation for targeted analysis:

  • Immunoprecipitate LTPG31 complexes for:

    • Lipidomics analysis of bound lipid cargo

    • Proteomics of interaction partners

    • Glycomics of associated cell wall fragments

  • Compare profiles across development and stress conditions

• Spatial transcriptomics with immunohistochemistry:

  • Correlate spatial gene expression patterns with protein localization

  • Map transcriptional domains to protein functional domains

  • Identify regulatory relationships through spatial correlation

Methodology integration table:

These integrative approaches leverage the specificity of antibodies while overcoming their limitations through complementary techniques, creating a more comprehensive understanding of LTPG31 biology than any single method could provide .

How might LTPG31 antibodies contribute to understanding evolutionary conservation of lipid transport mechanisms across plant species?

LTPG31 antibodies can serve as powerful tools for comparative evolutionary studies across plant lineages:

Cross-species immunodetection strategies:

• Epitope conservation analysis:

  • Perform multiple sequence alignments of LTPG family proteins across diverse plant species

  • Identify highly conserved epitopes within LTPG31 orthologs

  • Generate antibodies targeting these conserved regions

  • Test cross-reactivity against extracts from model and non-model species

  • Create conservation maps showing epitope preservation across plant phylogeny

• Comparative immunolocalization:

  • Apply optimized LTPG31 antibodies to tissues from diverse plant lineages

  • Compare subcellular localization patterns across evolutionary distance

  • Assess correlation between localization and specialized plant structures

  • Identify conserved vs. lineage-specific localization patterns

Evolutionary functional analysis approaches:

• Structure-function conservation assessment:

  • Immunoprecipitate LTPG31 orthologs from diverse species

  • Compare lipid binding profiles across evolutionary distance

  • Correlate functional conservation with sequence conservation

  • Identify critical regions through comparison of antibody epitope accessibility

• Heterologous complementation studies:

  • Express LTPG31 orthologs from diverse species in Arabidopsis ltpg31 mutants

  • Use antibodies to verify protein expression and localization

  • Assess functional complementation through physiological assays

  • Connect protein structure conservation to functional conservation

Evolutionary adaptation research:

• Environmental specialization analysis:

  • Compare LTPG31 expression and localization in plants from extreme environments

  • Correlate antibody-detected modifications with environmental adaptations

  • Identify specialized functions in drought, cold, or salt-adapted species

  • Map adaptive changes to specific protein domains

• Developmental program comparison:

  • Track LTPG31 expression during key developmental transitions across species

  • Compare timing and tissue-specificity of expression

  • Identify conserved developmental roles vs. species-specific functions

  • Relate to evolution of specialized structures (e.g., different cuticle types)

Methodological framework for cross-species antibody applications:

This evolutionary approach using antibodies has been successfully applied to study conservation of other plant protein families and can reveal how LTPG31 function has been conserved or diversified across plant evolution .