CESA5 Antibody

What is CESA5 and why are antibodies against it important in plant biology research?

CESA5 (Cellulose Synthase A5) is a key component of cellulose synthase complexes in plants that participates in primary cell wall synthesis. Research indicates that CESA5, along with CESA3 and CESA10, exhibits peak expression during mucilage biosynthesis, with both CESA3 and CESA10 showing higher expression levels than other CESAs during this developmental stage . Antibodies against CESA5 are crucial tools that enable researchers to:

• Detect and quantify CESA5 protein expression in different plant tissues and developmental stages

• Examine subcellular localization of CESA5 in different cell types

• Study protein-protein interactions between CESA5 and other cellulose synthase complex components

• Investigate the dynamics of cellulose synthase complex formation and movement

• Verify gene knockdown or knockout efficiency in functional genomics studies

The specificity of these antibodies is critical for distinguishing CESA5 from other closely related CESA family members, making them invaluable in dissecting the precise roles of individual CESAs in cell wall assembly.

How does CESA5 function differ from other CESA proteins?

CESA5 belongs to the cellulose synthase family but plays specialized roles that distinguish it from other CESA proteins:

• Specialized expression pattern: CESA5 shows elevated expression specifically during mucilage biosynthesis, working in concert with CESA3 and CESA10 .

• Protein interaction specificity: Research demonstrates that CESA5 interacts with CESA3 through its N-terminal zinc finger RING-type domain . This interaction appears to be highly specific and mediated through the cysteine-rich region of the RING domain.

• Complex assembly contributions: While proteins like CESA1, CESA3, and CESA6 form primary cell wall synthesis complexes in many tissues, CESA5 appears to have more specialized functions in certain developmental contexts.

• Structural features: CESA5 contains a distinctive N-terminal region with a Cys-rich RING-type zinc finger domain that mediates protein-protein interactions. When key cysteine residues (C39 and C84) are mutated to alanine, the protein loses its zinc-binding capacity and ability to maintain these interactions .

These functional differences make CESA5-specific antibodies essential for distinguishing its unique roles from those of other CESA proteins, particularly in specialized tissues where multiple CESA proteins are expressed simultaneously.

What are the key considerations for validating a CESA5 antibody?

Thorough validation of CESA5 antibodies is critical before their application in research. The validation process should include:

• Specificity testing: Verify that the antibody detects CESA5 without cross-reacting with other CESA family members. This is particularly important since CESA proteins share significant sequence homology. For example, in analyzing CESA antibodies, researchers should confirm lack of cross-reactivity with closely related isoforms (as seen with anti-CESA6 potentially cross-reacting with CESA2, CESA5, or CESA9) .

• Western blot validation: Confirm that the antibody detects a protein of the expected molecular weight (~120 kDa for most CESA proteins) . Validate using both native plant samples and recombinant CESA5 proteins as positive controls.

• Knockout/knockdown controls: Test the antibody on samples from cesa5 mutant/knockdown plants to confirm reduced or absent signal compared to wild-type samples.

• Immunoprecipitation efficiency: Assess the antibody's capability to immunoprecipitate CESA5 from plant protein extracts under both denaturing and non-denaturing conditions.

• Cross-species reactivity: Determine if the antibody can detect CESA5 orthologs in different plant species, which may be valuable for comparative studies. Methods like the Cross-species Epitope Sequence Analysis (CESA) can help predict antibody cross-reactivity across species .

• Epitope mapping: Identify the specific region of CESA5 recognized by the antibody, especially whether it targets the N-terminal region, catalytic domain, or C-terminus, as this impacts the detection of potential degradation products or splice variants.

How can CESA5 antibodies be used to study protein-protein interactions within cellulose synthase complexes?

CESA5 antibodies provide powerful tools for investigating the complex interactions between cellulose synthase components:

• Co-immunoprecipitation studies: CESA5 antibodies can be used in co-IP experiments to identify proteins that physically interact with CESA5. Similar to how CESA3 and CESA6 were shown to co-immunoprecipitate under non-denaturing conditions , CESA5 antibodies can help elucidate the composition of protein complexes containing CESA5.

• Comparison of interaction patterns in different conditions: Researchers can employ CESA5 antibodies to examine how protein-protein interactions change under different developmental stages, stress conditions, or in response to cell wall synthesis inhibitors.

• Domain-specific interaction analysis: By using CESA5 antibodies alongside mutated versions of CESA5 (such as those with mutations in the zinc finger domain), researchers can investigate which domains are critical for specific protein interactions. For example, studies have shown that mutations in the N-terminal cysteine residues of CESA5 (C39A and C84A) disrupt zinc binding and subsequent protein interactions .

• Proximity labeling approaches: CESA5 antibodies can be used in conjunction with techniques like BioID or APEX proximity labeling to identify proteins that are in close proximity to CESA5 in living cells, providing insights into the spatial organization of cellulose synthase complexes.

• Quantitative interaction analysis: When combined with techniques like surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC), purified CESA5 antibodies can help determine binding affinities and kinetics of interactions between CESA5 and its partners.

This detailed interaction analysis is crucial for understanding the assembly, regulation, and function of cellulose synthase complexes in different plant tissues and developmental contexts.

What approaches can be used to study CESA5 dynamics and trafficking using antibodies?

CESA5 antibodies enable sophisticated analyses of protein dynamics and trafficking within plant cells:

• Immunolocalization microscopy: Using CESA5-specific antibodies in combination with fluorescence microscopy to track the subcellular localization of CESA5 in fixed cells. This allows visualization of CESA5 distribution patterns in different cell types and developmental stages.

• Live-cell imaging with antibody fragments: Utilizing smaller antibody fragments (Fab fragments or nanobodies) conjugated to fluorophores to track CESA5 movement in living cells without significantly perturbing function.

• Pulse-chase experiments: Employing CESA5 antibodies in pulse-chase labeling experiments to track the turnover rate and half-life of CESA5 proteins, providing insights into protein stability and recycling dynamics.

• Endocytosis and trafficking studies: Using CESA5 antibodies to monitor the internalization and recycling of cellulose synthase complexes, similar to how GFP-tagged CESA proteins have been used to visualize bidirectional movement along microtubules .

• Velocity measurements: Combining antibody-based detection with time-lapse imaging to measure the movement velocities of CESA5-containing complexes at the cell surface. Previous studies with GFP-CESA3 and GFP-CESA6 have shown that these complexes migrate with comparable velocities (approximately 300-350 nm/min) .

• Colocalization analysis: Using dual-labeling approaches with CESA5 antibodies and markers for different cellular compartments to track the trafficking pathway of CESA5 through the secretory and endocytic pathways.

These approaches provide crucial insights into the dynamic behavior of CESA5 during cell wall synthesis and under different experimental conditions.

How can CESA5 antibodies be used to investigate post-translational modifications?

Post-translational modifications (PTMs) are critical regulatory mechanisms that influence CESA5 function and dynamics. CESA5 antibodies can be powerful tools for investigating these modifications:

• Phosphorylation-specific antibodies: Developing or using phospho-specific antibodies that recognize CESA5 only when phosphorylated at specific residues allows researchers to study how phosphorylation regulates CESA5 function. Cross-species analysis suggests that some phosphorylation sites may be conserved between species, with approximately 17% of human phosphosites conserved in Drosophila .

• PTM-dependent complex formation: Using standard CESA5 antibodies in conjunction with PTM-specific antibodies to investigate how modifications affect the formation and stability of cellulose synthase complexes.

• Immunoprecipitation followed by PTM analysis: Employing CESA5 antibodies to immunoprecipitate the protein, followed by mass spectrometry analysis to identify various PTMs and their stoichiometry under different conditions.

• In vitro reconstitution studies: Using purified CESA5 (obtained via immunoprecipitation with CESA5 antibodies) in in vitro systems to study how specific enzymes modify CESA5 and how these modifications affect its activity.

• Dynamic PTM changes: Tracking changes in CESA5 modifications during development or in response to environmental stresses using CESA5 antibodies in combination with PTM-specific detection methods.

• Mutational analysis validation: Using CESA5 antibodies to confirm the expression of mutated versions of CESA5 where potential PTM sites have been altered to either prevent modification (e.g., S→A mutations) or mimic constitutive modification (e.g., S→D mutations).

This approach is particularly powerful as it allows researchers to correlate specific modifications with functional outcomes in cellulose synthesis and cell wall formation.

What are the optimal protein extraction and sample preparation methods for CESA5 antibody applications?

Effective protein extraction and sample preparation are crucial for successful CESA5 antibody applications. Based on established protocols for CESA proteins:

• Extraction buffer optimization:

  • Use buffers containing 1% Triton X-100 for membrane protein solubilization, similar to protocols used for CESA3 and CESA6

  • Include protease inhibitor cocktails to prevent degradation

  • Add phosphatase inhibitors if studying phosphorylation states

  • Consider including reducing agents like DTT (1-5 mM) to maintain protein structure

• Tissue selection and processing:

  • Target tissues with known CESA5 expression (e.g., seed coat during mucilage biosynthesis)

  • Flash-freeze tissues in liquid nitrogen and grind to a fine powder

  • Extract proteins immediately or store powder at -80°C

  • Process samples consistently between experimental conditions to ensure comparability

• Protein quantification and normalization:

  • Use Bradford or BCA assays for protein quantification

  • Load equal amounts of total protein for immunoblotting (typically 10-30 μg)

  • Consider using purified recombinant CESA5 as a standard for absolute quantification

• Sample denaturation conditions:

  • For complete denaturation: heat samples at 95°C for 5 minutes in Laemmli buffer with SDS and β-mercaptoethanol

  • For native conditions: use non-denaturing buffers without heat treatment to preserve protein interactions

• Gel electrophoresis considerations:

  • Use 7-8% SDS-PAGE gels for optimal separation of large proteins like CESA5 (~120 kDa)

  • Consider gradient gels (4-15%) if analyzing CESA5 alongside interacting proteins of different sizes

  • Allow sufficient running time for proper separation of high molecular weight proteins

Following these optimized extraction and preparation methods will significantly improve the sensitivity and specificity of CESA5 detection in subsequent antibody-based applications.

What controls should be included in experiments using CESA5 antibodies?

Proper experimental controls are essential for ensuring the reliability and interpretability of results when using CESA5 antibodies:

• Positive controls:

  • Wild-type plant tissue known to express CESA5 (e.g., developing seed coats)

  • Recombinant CESA5 protein expressed in E. coli or other expression systems

  • Plants overexpressing CESA5 (if available)

• Negative controls:

  • cesa5 knockout or knockdown plant lines

  • Plant tissues known not to express CESA5

  • Primary antibody omission controls to assess secondary antibody non-specific binding

  • Pre-immune serum controls (if using polyclonal antibodies)

  • Blocking peptide controls where the primary antibody is pre-incubated with the antigen used for immunization

• Specificity controls:

  • Testing against recombinant proteins from related CESA family members to ensure no cross-reactivity

  • Immunoblotting against protein extracts from mutants of related CESA proteins

  • Testing the antibody against the N-terminal fragments of various CESA proteins produced in E. coli, similar to validation methods used for other CESA antibodies

• Method-specific controls:

  • For co-immunoprecipitation: denaturing vs. non-denaturing conditions to differentiate direct interactions from complex associations

  • For immunolocalization: peptide competition assays to confirm signal specificity

  • For quantitative western blotting: loading controls (e.g., actin, tubulin) and standard curves

• Cross-reactivity testing:

  • If working with multiple plant species, validate the antibody's reactivity in each species

  • Use tools like Cross-species Epitope Sequence Analysis (CESA) to predict antibody cross-reactivity

Proper implementation of these controls will significantly enhance the reliability of experiments using CESA5 antibodies and facilitate accurate interpretation of results across different experimental contexts.

What are the considerations for developing new CESA5 antibodies for specific research applications?

Developing new CESA5 antibodies requires careful planning to ensure specificity, sensitivity, and suitability for intended applications:

• Epitope selection strategy:

  • Choose unique regions of CESA5 with minimal homology to other CESA family members

  • Consider the N-terminal region (amino acids 1-262), which contains the zinc finger domain involved in protein-protein interactions

  • Avoid transmembrane domains, which may be inaccessible in native proteins

  • Use epitope prediction algorithms to identify regions likely to be surface-exposed and immunogenic

• Antigen preparation approaches:

  • Express recombinant fragments of CESA5 as GST-fusion proteins in E. coli, similar to methods used for CESA3 and CESA6 antibodies

  • Purify using affinity chromatography (e.g., GST-Sepharose 4B resin)

  • Verify correct folding of important structural elements (e.g., zinc binding in RING domains) using appropriate assays

• Immunization and antibody production protocols:

  • Use rabbits for polyclonal antibodies or mice for monoclonal antibody development

  • Implement a multi-step immunopurification procedure to enhance specificity

  • Consider developing antibodies against post-translationally modified forms of CESA5

• Validation for specific applications:

  • Western blotting: Test against wild-type and cesa5 mutant tissues

  • Immunoprecipitation: Verify ability to precipitate CESA5 from plant extracts

  • Immunolocalization: Confirm specific subcellular localization patterns

  • Cross-species reactivity: Test against CESA5 orthologs in related plant species

• Specialized antibody formats:

  • Consider developing recombinant antibody fragments (Fab, scFv) for applications requiring smaller antibody size

  • Explore nanobody development for live-cell imaging applications

  • Develop directly conjugated antibodies (fluorophore, enzyme) for specific applications

Thoughtful consideration of these factors will lead to the development of CESA5 antibodies optimally suited for specific research objectives and experimental conditions.

How should researchers interpret unexpected CESA5 antibody results?

When using CESA5 antibodies, researchers may encounter unexpected results that require careful interpretation:

• Multiple bands in Western blots:

  • Possible interpretation: Presence of alternative splice variants, degradation products, or post-translationally modified forms of CESA5

  • Similar phenomenon: Anti-CESA6 antibodies have been observed to detect a second band that may correspond to a truncated CESA6 isoform or a CESA6-related protein

  • Validation approach: Compare with known molecular weights, confirm with mass spectrometry, and test in mutant backgrounds

• Reduced CESA5 signal in wild-type samples:

  • Possible interpretation: Co-regulation with other CESA proteins, similar to how mutations in CESA1, CESA3, or CESA6 can cause decreased protein levels of CESA3 and CESA6

  • Verification strategy: Examine transcript levels by RT-PCR and protein stability under different extraction conditions

• Unexpected subcellular localization:

  • Possible interpretation: Novel trafficking pathways, stress response relocalization, or detection of a subpopulation of CESA5

  • Confirmation approach: Use multiple antibodies targeting different epitopes of CESA5 and compare with fluorescently tagged CESA5 localization

• Cross-reactivity with other proteins:

  • Possible interpretation: Antibody recognizing conserved epitopes in related proteins

  • Similar issue: Anti-CESA6 antibodies potentially cross-reacting with closely related isoforms CESA2, CESA5, or CESA9

  • Resolution strategy: Perform immunoprecipitation followed by mass spectrometry to identify all recognized proteins

• Differential detection across developmental stages:

  • Possible interpretation: Developmental regulation of CESA5 expression, similar to how CESA3 and CESA6 were undetectable in imbibed seeds and appeared after germination

  • Analysis approach: Conduct time-course studies of protein expression and correlate with known developmental events

Understanding these potential scenarios and their biological implications will help researchers accurately interpret their CESA5 antibody results and design appropriate follow-up experiments to resolve ambiguities.

What are common technical issues with CESA5 antibodies and how can they be resolved?

Researchers may encounter several technical challenges when working with CESA5 antibodies. Here are common issues and their solutions:

• Low signal intensity:

  • Problem: Insufficient CESA5 protein in samples or low antibody sensitivity

  • Solutions:

    • Optimize protein extraction with appropriate detergents (e.g., 1% Triton X-100)

    • Increase antibody concentration or incubation time

    • Use signal enhancement systems (e.g., biotin-streptavidin amplification)

    • Target tissues with known high CESA5 expression (e.g., seed coat during mucilage biosynthesis)

• High background:

  • Problem: Non-specific antibody binding or inadequate blocking

  • Solutions:

    • Implement a multi-step immunopurification procedure for antibodies, similar to methods used for CESA3 and CESA6 antibodies

    • Optimize blocking conditions (try different blocking agents like 5% BSA or 5% milk)

    • Include additional washing steps with higher salt concentration or detergent

    • Consider using more specific secondary antibodies or monoclonal primary antibodies

• Inconsistent immunoprecipitation efficiency:

  • Problem: Variable pulling down of CESA5 in different experiments

  • Solutions:

    • Standardize extraction conditions (buffer composition, temperature, incubation time)

    • Pre-clear lysates with protein A/G beads before immunoprecipitation

    • Crosslink antibody to beads to prevent antibody contamination in eluted samples

    • Optimize detergent concentration to solubilize membrane proteins without disrupting interactions

• Poor reproducibility between experiments:

  • Problem: Batch-to-batch variation in antibodies or sample preparation

  • Solutions:

    • Purchase larger lots of antibodies and aliquot to minimize freeze-thaw cycles

    • Standardize tissue collection and protein extraction protocols

    • Include internal controls in each experiment

    • Consider switching to monoclonal antibodies for greater consistency

• Degradation of CESA5 during sample preparation:

  • Problem: Proteolytic breakdown leading to multiple bands or weak signal

  • Solutions:

    • Add protease inhibitor cocktails to all buffers

    • Keep samples cold throughout processing

    • Process samples quickly and avoid repeated freeze-thaw cycles

    • Consider using fresher tissue samples

Implementing these technical solutions will help researchers overcome common challenges and obtain more reliable and reproducible results when using CESA5 antibodies in their experimental work.

How can CESA5 antibody studies be combined with genetic approaches for comprehensive analysis?

Integrating CESA5 antibody-based studies with genetic approaches provides powerful insights into cellulose synthase function and regulation:

• Complementary analysis of mutant lines:

  • Use CESA5 antibodies to analyze protein levels in various cesa mutants to uncover compensatory mechanisms

  • Similar to studies showing reduced CESA3 and CESA6 levels in cesa1, cesa3, and cesa6 mutants , examine how mutations in other CESA genes affect CESA5 protein levels

  • Compare protein expression patterns with phenotypic outcomes to establish structure-function relationships

• CRISPR/Cas9 genome editing validation:

  • Employ CESA5 antibodies to confirm protein knockout or modification following genome editing

  • Use immunoblotting to detect truncated or modified CESA5 proteins resulting from genome editing

  • Correlate editing efficiency at the DNA level with protein expression changes

• Protein-level analysis of RNAi or amiRNA knockdowns:

  • Quantify the extent of CESA5 protein reduction following transcript-level knockdown

  • Determine the correlation between transcript reduction and protein reduction

  • Assess whether partial protein reduction leads to intermediate phenotypes

• Transgenic complementation studies:

  • Use CESA5 antibodies to confirm expression of transgenic CESA5 variants in mutant backgrounds

  • Quantify expression levels of complementing proteins relative to endogenous CESA5 in wild-type plants

  • Correlate transgene expression levels with functional recovery in complementation experiments

• Domain function analysis:

  • Analyze the expression and stability of CESA5 proteins with specific domain mutations or deletions

  • Similar to studies with CESA5 N-terminal zinc finger domain mutants (C39A and C84A) , use antibodies to confirm protein expression and investigate functional consequences

  • Investigate how domain mutations affect interactions with other CESA proteins

This integrated approach leverages the strengths of both genetic manipulation and protein-level analysis to provide a more complete understanding of CESA5 function in cellulose synthesis and plant development.

What mass spectrometry approaches can be combined with CESA5 immunoprecipitation for advanced proteomics?

Combining CESA5 immunoprecipitation with mass spectrometry creates powerful workflows for in-depth proteomic analysis:

• Interaction proteomics workflow:

  • Immunoprecipitate CESA5 using validated antibodies under non-denaturing conditions

  • Analyze co-precipitated proteins using LC-MS/MS to identify interacting partners

  • Compare results between different developmental stages or treatments to identify condition-specific interactions

  • Differentiate between stable complex components and transient interactors using quantitative approaches

• Post-translational modification mapping:

  • Immunoprecipitate CESA5 using specific antibodies

  • Digest purified CESA5 with proteases and analyze peptides by LC-MS/MS

  • Implement neutral loss scanning for phosphorylation or precursor ion scanning for glycosylation

  • Compare modification patterns across different conditions to identify regulatory PTMs

• Cross-linking mass spectrometry (XL-MS):

  • Apply protein cross-linkers to intact cells or isolated complexes prior to CESA5 immunoprecipitation

  • Analyze cross-linked peptides by specialized MS workflows to identify proximity relationships

  • Generate structural constraints for modeling CESA5 complexes

  • Map interaction interfaces between CESA5 and other cellulose synthase components

• Absolute quantification of CESA5:

  • Develop selected reaction monitoring (SRM) or parallel reaction monitoring (PRM) assays for CESA5-specific peptides

  • Use stable isotope-labeled peptide standards for absolute quantification

  • Apply targeted proteomics to measure CESA5 abundance across different tissues or treatments

• Comparative proteomics between wild-type and mutant backgrounds:

  • Immunoprecipitate CESA5-containing complexes from wild-type and various cesa mutant backgrounds

  • Identify differential complex composition using label-free quantification or isobaric labeling (TMT, iTRAQ)

  • Determine how complex assembly is affected by the absence of specific components

These mass spectrometry approaches extend the analytical power of CESA5 antibodies beyond traditional applications, enabling researchers to answer increasingly sophisticated questions about CESA5 biology and function.

What are emerging technologies and future directions for CESA5 antibody applications?

As research on plant cell wall biosynthesis advances, several emerging technologies are poised to enhance CESA5 antibody applications:

• Super-resolution microscopy:

  • Application of techniques like STORM, PALM, or STED microscopy with CESA5 antibodies to visualize cellulose synthase complex organization below the diffraction limit

  • Potential to resolve individual CESA5-containing complexes within the plasma membrane and track their movement with nanometer precision

  • Opportunity to correlate complex organization with nascent cellulose microfibril deposition

• Single-molecule antibody approaches:

  • Development of methods to track individual CESA5 molecules using antibody fragments

  • Analysis of stoichiometry and dynamics at the single-molecule level

  • Investigation of heterogeneity in complex composition and behavior

• Cryo-electron microscopy with antibody labeling:

  • Use of CESA5 antibodies or antibody fragments as fiducial markers for cryo-EM structural studies

  • Potential to determine the structural organization of CESA5 within cellulose synthase complexes

  • Integration with in situ structural biology approaches to visualize complexes in their native environment

• Cross-species antibody development using CESA technology:

  • Application of Cross-species Epitope Sequence Analysis (CESA) to develop antibodies that work across multiple plant species

  • Enabling comparative studies of CESA5 function in diverse plant lineages

  • Potential to identify conserved and divergent aspects of cellulose synthase complex organization

• Spatially-resolved proteomics:

  • Combination of CESA5 antibodies with emerging spatial proteomics techniques to map protein interactions in specific subcellular locations

  • Integration with proximity labeling approaches to identify context-specific interactors

  • Development of multiplexed antibody-based imaging to simultaneously track multiple CESA proteins

• Therapeutic antibody-inspired approaches:

  • Creation of antibodies that modulate CESA5 function rather than simply detecting it

  • Development of antibody-based tools to manipulate cellulose synthesis in controlled ways

  • Application of antibody engineering principles to create highly specific tools for plant cell biology

These emerging directions represent exciting opportunities for researchers to gain deeper insights into CESA5 biology and cellulose synthesis mechanisms using increasingly sophisticated antibody-based approaches.

What are the key challenges and opportunities in CESA5 antibody research?

The field of CESA5 antibody research faces several challenges but also offers significant opportunities for advancement:

Challenges:

• Specificity limitations: Distinguishing CESA5 from other closely related CESA family members remains difficult due to high sequence homology. Similar challenges have been noted with anti-CESA6 antibodies potentially cross-reacting with CESA2, CESA5, or CESA9 .

• Membrane protein complexes: CESA5 exists in large membrane-embedded complexes, making extraction and preservation of native structure challenging for antibody-based studies.

• Post-translational modification diversity: The various PTMs on CESA5 may affect antibody recognition, leading to incomplete detection of all CESA5 protein states.

• Limited commercial availability: Unlike antibodies for common mammalian proteins, plant-specific CESA5 antibodies have limited commercial availability and often require in-house development.

• Cross-species application: Extending antibody applications across diverse plant species requires extensive validation, though tools like CESA (Cross-species Epitope Sequence Analysis) are emerging to address this challenge .

Opportunities:

• Integration with emerging technologies: Combining CESA5 antibodies with advances in super-resolution microscopy, cryo-EM, and mass spectrometry offers unprecedented insights into cellulose synthase structure and function.

• Synthetic biology applications: As plant cell wall engineering becomes more sophisticated, CESA5 antibodies will be essential tools for validating and characterizing engineered systems.

• Environmental response studies: Using CESA5 antibodies to study how cellulose synthesis responds to environmental stresses could lead to more resilient crop developments.

• Cross-disciplinary applications: The methods developed for CESA5 antibody production and validation could inform approaches for other challenging plant membrane proteins.

• Therapeutic agricultural applications: Knowledge gained from CESA5 antibody studies could potentially inform the development of agents that modulate cell wall biosynthesis for agricultural improvements.