At5g44390 Antibody

What is At5g44390 and why is it important in plant research?

At5g44390 (UniProt: Q9FKU9) is a protein in Arabidopsis thaliana that belongs to the Berberine bridge enzyme-like family (BBE-like 25, AtBBE-like 25). It is classified within the oxygen-dependent FAD-linked oxidoreductase family and is primarily localized to the cell wall as a secreted protein. The protein plays roles in plant defense mechanisms and secondary metabolism pathways, making it an important target for studying plant response to various environmental stressors. Research on At5g44390 contributes to our understanding of plant biochemical pathways involved in development and stress responses, which has implications for improving crop resilience and productivity.

What applications can the At5g44390 antibody be used for?

The At5g44390 antibody has been tested and validated for several research applications:

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative detection of At5g44390 protein in complex biological samples .

• Western Blotting (WB): For identification and semi-quantitative analysis of At5g44390 protein expression in plant tissue extracts .

Both applications allow researchers to study protein expression patterns, tissue distribution, and changes in protein levels under different experimental conditions. When designing experiments, it's essential to optimize protocols specifically for plant tissue extracts, as the cellular composition differs significantly from animal tissues. Standard blocking solutions containing 3-5% BSA or non-fat milk in TBST buffer typically work well for reducing background signal in these applications.

How should the At5g44390 antibody be stored and handled for optimal performance?

For optimal antibody performance and longevity, follow these storage and handling recommendations:

• Upon receipt, store the antibody at -20°C or -80°C for long-term storage .

• Avoid repeated freeze-thaw cycles that can lead to protein denaturation and loss of antibody activity .

• The antibody is supplied in a liquid form containing preservative (0.03% Proclin 300) and stabilizers (50% Glycerol, 0.01M PBS, pH 7.4) .

• When working with the antibody, keep it on ice and return to storage promptly after use.

• For Western blotting applications, dilution ratios typically range from 1:1000 to 1:5000, but optimal dilutions should be determined empirically for your specific experimental conditions.

Proper storage and handling practices are critical for maintaining antibody specificity and sensitivity, particularly for plant-specific antibodies that may have limited commercial availability and long lead times (14-16 weeks for At5g44390 antibody) .

What controls should be included when using the At5g44390 antibody?

When designing experiments with the At5g44390 antibody, include the following controls to ensure reliable and interpretable results:

• Positive Control: Use wild-type Arabidopsis thaliana tissue samples known to express At5g44390 protein. Root or leaf extracts from Col-0 ecotype plants grown under standard conditions often provide suitable positive controls.

• Negative Control: Include samples from:

  • At5g44390 knockout mutant plants (if available)

  • Non-plant tissue or unrelated protein samples

  • Primary antibody omission control (to assess secondary antibody specificity)

• Loading Control: For Western blot experiments, include detection of a housekeeping protein such as actin, tubulin, or GAPDH to normalize for loading variations.

• Specificity Controls: Pre-absorption of the antibody with the immunizing peptide/recombinant protein can confirm binding specificity. This approach is particularly valuable when antibody cross-reactivity is suspected.

Including these controls helps validate experimental findings and addresses potential concerns about antibody specificity, which is crucial when submitting findings for peer review or publication .

How can I optimize Western blotting protocols for detecting At5g44390 in different plant tissues?

Optimizing Western blotting protocols for At5g44390 detection requires careful consideration of several experimental parameters:

• Extraction Buffer Composition:

  • For cell wall-associated proteins like At5g44390, use extraction buffers containing:

    • 50 mM Tris-HCl (pH 8.0)

    • 150 mM NaCl

    • 1% Triton X-100

    • Protease inhibitor cocktail

    • Consider adding 2-5% β-mercaptoethanol to reduce disulfide bonds

• Tissue-Specific Considerations:

  Tissue Type	Recommended Modifications	Expected Protein Yield
  Leaf	Add 2% PVPP to remove phenolic compounds	Moderate
  Root	Increase grinding time; add 0.5% SDS	Variable
  Flowers	Use gentler homogenization; add 5 mM EDTA	Low-Moderate
  Seeds	Extended extraction time; add 4M urea	Low

• Transfer Optimization:

  • For secreted proteins like At5g44390, semi-dry transfer systems with 15% methanol in transfer buffer often yield better results

  • Transfer at 15V for 45-60 minutes for proteins between 30-70 kDa

• Detection System Selection:

  • Chemiluminescent detection offers good sensitivity for plant proteins expressed at low levels

  • Consider using HRP-conjugated secondary antibodies with enhanced chemiluminescent substrates

• Blocking Parameters:

  • Test both 5% non-fat milk and 3% BSA in TBST to determine optimal blocking conditions

  • Extended blocking times (2-3 hours at room temperature or overnight at 4°C) may reduce background

These optimizations address the challenges associated with extracting and detecting cell wall-associated proteins like At5g44390, which can be difficult to solubilize and may require specialized extraction protocols .

What approaches can be used to study At5g44390 protein interactions with other cellular components?

Investigating At5g44390 protein interactions requires specialized techniques addressing the challenges of studying cell wall-localized proteins:

• Co-Immunoprecipitation (Co-IP):

  • Use crosslinking agents like formaldehyde (1-2%) or DSP (dithiobis[succinimidyl propionate]) to stabilize transient interactions

  • Extract protein complexes using optimized buffer systems containing:

    • 50 mM HEPES (pH 7.5)

    • 150 mM NaCl

    • 1 mM EDTA

    • 0.5% NP-40

    • Protease inhibitor cocktail

  • Immunoprecipitate with At5g44390 antibody bound to Protein A/G beads

  • Analyze co-precipitated proteins by mass spectrometry

• Proximity-Based Labeling:

  • Generate fusion constructs of At5g44390 with BioID or TurboID biotin ligase

  • Express in Arabidopsis to biotinylate proximal proteins

  • Purify biotinylated proteins using streptavidin beads

  • This approach is particularly useful for studying cell wall protein interactions

• Yeast Two-Hybrid (Y2H) Screening:

  • Despite limitations for secreted proteins, using truncated versions lacking signal peptides can identify potential cytoplasmic interaction partners

  • Split-ubiquitin Y2H systems may be more appropriate for membrane-proximal interactions

• Bimolecular Fluorescence Complementation (BiFC):

  • Generate fusion constructs of At5g44390 and candidate interacting proteins with split fluorescent protein fragments

  • Transiently express in Arabidopsis protoplasts or Nicotiana benthamiana leaves

  • Visualize protein interactions through fluorescence microscopy

When interpreting results, consider that as a secreted protein localized to the cell wall, At5g44390 interactions may be transient or dependent on specific cellular conditions related to plant defense or secondary metabolism pathways.

How can I design experiments to investigate At5g44390 function using both antibody-based detection and genetic approaches?

A comprehensive strategy to investigate At5g44390 function should combine antibody-based detection methods with genetic manipulation approaches:

• Experimental Design Framework:

  Approach	Method	Purpose	Key Controls
  Antibody-based	Immunohistochemistry	Tissue localization	Secondary antibody only; At5g44390 knockout
  Antibody-based	Protein expression analysis	Expression patterns under stress	Loading controls; time-course samples
  Genetic	T-DNA insertion lines	Loss-of-function analysis	Wild-type; complementation lines
  Genetic	CRISPR/Cas9 editing	Precise functional domain analysis	Off-target analysis; wild-type
  Combined	Complementation + antibody detection	Functional domain mapping	Empty vector; wild-type protein expression

• Experimental Procedure Integration:

  • Begin with phenotypic characterization of At5g44390 knockout/knockdown lines under various conditions

  • Use the antibody to confirm protein absence in mutant lines and quantify expression in wild-type plants

  • Complement mutant lines with native or modified At5g44390 constructs

  • Analyze protein expression, localization, and function in complemented lines using the antibody

• Oxidoreductase Activity Assessment:
  As At5g44390 belongs to the oxygen-dependent FAD-linked oxidoreductase family, design specific enzyme activity assays:

  • Measure enzyme kinetics using purified native protein or recombinant protein

  • Assess FAD binding using spectrophotometric methods

  • Compare activity in wild-type extracts versus knockout lines

  • Use the antibody to immunodeplete the protein and test for loss of specific enzymatic activities

• Stress Response Studies:

  • Subject plants to various stressors (pathogen infection, drought, salinity)

  • Monitor At5g44390 protein levels using the antibody via Western blotting

  • Compare phenotypic responses between wild-type and knockout plants

  • Perform transcriptome analysis to identify genes co-regulated with At5g44390

This integrated approach combines the specificity of antibody-based detection with the functional insights provided by genetic manipulation, offering a more comprehensive understanding of At5g44390's biological role .

What troubleshooting approaches can address common issues when using the At5g44390 antibody in plant tissue analyses?

When encountering problems with At5g44390 antibody applications, systematic troubleshooting can help identify and resolve issues:

• Low or No Signal in Western Blot:

  • Increase antibody concentration (try 1:500 instead of 1:1000)

  • Optimize protein extraction using cell wall-specific extraction buffers containing detergents suitable for secreted proteins

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

  • Enhance detection sensitivity with amplified chemiluminescent substrates

  • Verify target protein expression in your specific experimental conditions

  • Check protein transfer efficiency using reversible total protein stains

• High Background:

  • Increase blocking time and concentration (5% BSA for 2 hours)

  • Add 0.1-0.3% Tween-20 to wash buffers

  • Perform additional washing steps (5-6 washes, 10 minutes each)

  • Try alternative blocking agents (casein, commercial blocker formulations)

  • Decrease secondary antibody concentration

  • Pre-absorb primary antibody with plant extract lacking At5g44390

• Multiple Bands or Unexpected Band Size:

  • Consider post-translational modifications (At5g44390 may undergo glycosylation)

  • Test reducing vs. non-reducing conditions to assess disulfide bonding

  • Evaluate protein degradation by adding additional protease inhibitors

  • Use freshly prepared samples and avoid freeze-thaw cycles

  • Optimize gel percentage to better resolve proteins in your target's molecular weight range

• Variability Between Experiments:

  • Standardize protein extraction protocols rigorously

  • Use the same antibody lot number when possible

  • Implement quantitative loading controls

  • Develop a standard curve using recombinant At5g44390 protein

  • Document all experimental conditions meticulously

• Cross-Reactivity Concerns:

  • Validate results using genetic knockouts of At5g44390

  • Perform peptide competition assays using the immunizing antigen

  • Consider testing the antibody on other plant species to assess specificity

  • Use alternative antibody preparations (C-terminal vs. N-terminal) if available

Each troubleshooting approach should be documented systematically, with one variable modified at a time to identify the specific source of the issue .

How should I design experiments to study At5g44390 expression patterns during plant development and stress responses?

To effectively study At5g44390 expression patterns across development and stress conditions, implement a multi-faceted experimental design:

• Developmental Expression Analysis:

  • Collect tissue samples at key developmental stages:

    • Seedling (3, 7, and 14 days)

    • Vegetative growth (rosette leaves at different positions)

    • Reproductive transition (inflorescence emergence)

    • Flowering (flowers at stages 1-12)

    • Silique development (early, mid, mature)

    • Senescence

  • Process all samples in parallel using standardized protein extraction protocols

  • Perform Western blot analysis with At5g44390 antibody, including appropriate loading controls

  • Quantify relative protein abundance across developmental stages

• Stress Response Experimental Design:

  Stress Type	Treatment Conditions	Sampling Timepoints	Controls
  Drought	Withhold water for 3, 7, 10 days	0, 3, 7, 10 days	Well-watered plants
  Salt	150 mM NaCl solution	0, 6, 12, 24, 48 hours	Water-treated plants
  Cold	4°C exposure	0, 6, 12, 24, 48 hours	Plants at 22°C
  Pathogen	P. syringae infiltration	0, 12, 24, 48, 72 hours	Mock-infiltrated plants
  Wounding	Mechanical damage	0, 1, 3, 6, 24 hours	Unwounded plants

• Tissue-Specific Expression:

  • Use both protein extraction followed by Western blotting and immunohistochemistry with the At5g44390 antibody

  • For immunohistochemistry, prepare thin sections of different tissues embedded in paraffin or resin

  • Include appropriate negative controls (primary antibody omission, pre-immune serum)

  • Compare protein localization patterns with published transcriptomic data

• Genetic Background Comparisons:

  • Analyze At5g44390 expression in different Arabidopsis ecotypes (Col-0, Ws, Ler)

  • Include known mutants affecting pathways potentially related to At5g44390 function

  • Create transgenic lines with promoter-reporter constructs to complement antibody-based detection

• Variables to Control:

  • Standardize growth conditions (light intensity, photoperiod, temperature, humidity)

  • Harvest tissues at the same time of day to control for circadian effects

  • Use plants of identical age for stress treatments

  • Process all samples using identical extraction and detection protocols

This comprehensive experimental design employs the At5g44390 antibody as a key tool while incorporating appropriate controls and multiple analytical approaches to produce robust, reproducible findings about protein expression patterns .

What are the best practices for quantifying At5g44390 protein levels in comparative studies?

Accurate quantification of At5g44390 protein levels requires rigorous methodology to ensure comparability across samples and experiments:

• Standardized Protein Extraction Protocol:

  • For cell wall-localized proteins like At5g44390, use sequential extraction:

    • First extract: 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% Triton X-100

    • Second extract: Add 1% SDS to remaining pellet

    • Pool extracts or analyze separately depending on research question

  • Process all samples simultaneously under identical conditions

  • Use consistent sample-to-buffer ratios (e.g., 100 mg tissue per 300 μl buffer)

• Quantification Methodology:

  • Determine total protein concentration using Bradford or BCA assay

  • Prepare standard curves with each assay to ensure linearity within your sample range

  • Load equal amounts of total protein (20-50 μg) for all samples

  • Include gradient standards of recombinant At5g44390 protein (if available) for absolute quantification

• Western Blot Optimization for Quantification:

  • Use PVDF membranes for better protein retention and quantification linearity

  • Optimize antibody concentration to ensure signal remains in linear range

  • Employ digital imaging systems with appropriate dynamic range

  • Use technical replicates (multiple lanes of same sample) to assess technical variability

• Data Analysis Framework:

  Quantification Method	Advantages	Limitations	Software Tools
  Normalized band intensity	Simple, widely accepted	Limited dynamic range	ImageJ, Image Lab
  Density ratio to loading control	Controls for loading variation	Assumes constant expression of reference protein	ImageJ with Analyze Gels function
  Standard curve method	Provides absolute quantification	Requires purified recombinant protein	Excel, GraphPad Prism
  Multiplex fluorescent detection	Simultaneous target and control detection	Requires specialized equipment	LI-COR Image Studio

• Statistical Analysis:

  • Perform experiments with at least 3 biological replicates

  • Use appropriate statistical tests based on data distribution (t-test, ANOVA)

  • Report both fold-changes and absolute values when possible

  • Include error bars representing standard deviation or standard error

  • Calculate and report p-values for statistical significance

• Validation Approaches:

  • Confirm key findings using alternative methods (e.g., ELISA, immunoprecipitation)

  • Consider using mass spectrometry-based quantification for critical comparisons

  • Correlate protein levels with mRNA expression data when available

These best practices ensure that quantitative comparisons of At5g44390 protein levels are scientifically sound and reproducible across different experimental conditions .

How can I differentiate between specific and non-specific binding when using the At5g44390 antibody?

Distinguishing specific from non-specific binding is critical for accurate data interpretation when using the At5g44390 antibody:

• Validation Controls:

  • Genetic knockout validation: Compare Western blot signals between wild-type and confirmed At5g44390 knockout lines

  • Peptide competition assay: Pre-incubate antibody with excess immunizing peptide before application

  • Antibody dilution series: Specific signals typically maintain relative intensity across dilutions while non-specific signals often diminish disproportionately

• Signal Characteristics Analysis:

  Signal Characteristic	Likely Specific Binding	Potential Non-specific Binding
  Band molecular weight	Matches predicted size (± post-translational modifications)	Multiple random bands or major bands at unexpected sizes
  Signal consistency	Reproducible across experiments	Variable appearance between replicates
  Background pattern	Clean background with minimal additional bands	Smeared or ladder-like pattern
  Response to blocking	Maintained signal with increased blocking	Reduced signal with increased blocking
  Tissue specificity	Follows expected biological distribution	Appears uniformly across all tissues

• Technical Approaches:

  • Use two different antibodies targeting separate epitopes of At5g44390

  • Compare polyclonal (recognizes multiple epitopes) to monoclonal (single epitope) antibodies if available

  • Test antibody specificity on dot blots with purified recombinant At5g44390 protein

  • Perform immunoprecipitation followed by mass spectrometry to confirm identity of detected proteins

• Confounding Factors to Consider:

  • Cross-reactivity with related Berberine bridge enzyme-like family members

  • Alternative splice variants or post-translational modifications affecting antibody recognition

  • Protein degradation products generating fragments detected by the antibody

  • High protein concentration effects leading to non-specific interactions

• Optimized Blocking Strategy:

  • Test multiple blocking agents (BSA, non-fat milk, commercial blockers)

  • Add 0.1-0.2% Tween-20 to reduce hydrophobic interactions

  • Consider adding 5% normal serum from the secondary antibody host species

  • For plant tissues specifically, add 1% polyvinylpyrrolidone to reduce phenolic compound interference

By systematically implementing these approaches, researchers can confidently distinguish specific At5g44390 detection from non-specific antibody interactions, improving data reliability and interpretation .

How should I interpret conflicting results between antibody-based detection and transcript analysis of At5g44390?

Discrepancies between protein detection using the At5g44390 antibody and transcript analysis (RT-PCR or RNA-seq) are common in biological research and require careful interpretation:

• Biological Explanations for Discrepancies:

  • Post-transcriptional regulation: mRNA may be transcribed but not efficiently translated

  • Protein stability differences: Protein may have longer/shorter half-life than its mRNA

  • Temporal delay: Protein accumulation typically lags behind transcript induction

  • Tissue-specific translational control: mRNA may be present but translated only in specific contexts

  • Subcellular localization: As a secreted protein, At5g44390 may be difficult to extract completely

• Technical Considerations:

  Factor	Impact on Protein Detection	Impact on Transcript Detection
  Extraction efficiency	Cell wall proteins require specialized extraction	RNA extraction typically more standardized
  Detection sensitivity	Western blot may miss low abundance proteins	qRT-PCR can detect low-copy transcripts
  Quantification range	Often narrower dynamic range	Typically wider dynamic range
  Specificity	Potential cross-reactivity with related proteins	Primer design affects specificity
  Sample preparation	Protein degradation during extraction	RNA degradation during extraction

• Resolution Strategies:

  • Temporal analysis: Sample at multiple timepoints to detect potential delays between transcription and translation

  • Use multiple antibodies targeting different epitopes of At5g44390

  • Implement polysome profiling to assess translational status of At5g44390 mRNA

  • Perform absolute quantification of both transcript and protein

  • Create reporter gene fusions to monitor protein stability

• Integrated Data Analysis Approach:

  • Calculate protein-to-mRNA ratios across conditions to identify regulatory patterns

  • Compare fold-changes rather than absolute values between transcript and protein

  • Examine correlation patterns within specific tissue types or treatments

  • Consider proteome-wide analyses to determine if discrepancy is specific to At5g44390 or represents a broader cellular response

• Experimental Validation:

  • Generate transgenic plants expressing epitope-tagged At5g44390 under its native promoter

  • Use both antibody-based detection and reporter-based visualization

  • Employ cell-fractionation approaches to ensure complete extraction

  • Consider protein degradation inhibitors to assess turnover rates

When reporting these discrepancies in publications, clearly describe both transcript and protein detection methodologies, acknowledge limitations, and propose biological mechanisms that might explain the observed differences .

How can the At5g44390 antibody be utilized in chromatin immunoprecipitation (ChIP) experiments to study protein-DNA interactions?

While At5g44390 is a secreted cell wall protein and not typically expected to interact directly with DNA, there are scenarios where ChIP experiments might be valuable:

• Experimental Design Considerations:

  • Generate epitope-tagged versions of At5g44390 (FLAG, HA, or MYC tags) expressed under native promoter

  • Use native At5g44390 antibody or commercial anti-epitope antibodies for immunoprecipitation

  • Implement formaldehyde crosslinking (1-2%, 10-15 minutes) to capture potential transient interactions

  • Include appropriate controls (non-specific IgG, input chromatin)

• Protocol Optimization for Plant Tissues:

  • Modify standard ChIP protocols to accommodate plant cell wall complexities:

    • Extended nuclei isolation steps with additional grinding

    • Increase crosslinking time slightly (15-20 minutes) compared to standard protocols

    • Use sonication parameters optimized for plant chromatin (typically requiring longer sonication times)

  • Test different extraction buffers optimized for cell wall proteins

• Application Scenarios:

  Research Question	Experimental Approach	Controls Needed	Expected Outcome
  Indirect DNA association through protein complexes	Sequential ChIP (Re-ChIP)	Single ChIP controls	Enrichment of specific genomic regions
  Stress-induced nuclear translocation	Compare ChIP under normal vs. stress conditions	Cellular fractionation validation	Differential binding patterns under stress
  Involvement in chromatin remodeling pathways	ChIP followed by sequencing (ChIP-seq)	IgG ChIP-seq, input controls	Genome-wide binding profile

• Data Analysis Considerations:

  • Use peak-calling algorithms designed for plant ChIP-seq data

  • Implement more stringent filtration criteria due to potential non-specific binding

  • Compare binding profiles with RNA-seq data to identify potential regulatory relationships

  • Validate key findings with independent methods (e.g., EMSA, DNA-protein pulldown)

• Potential Limitations:

  • As a primarily secreted protein, nuclear localization may be limited or context-dependent

  • Higher background may be expected compared to typical transcription factor ChIP

  • Protein abundance in nuclear fraction may be limiting factor

  • Cross-reactivity with related BBE-like proteins could confound results

• Alternative Approaches:

  • Consider DamID (DNA adenine methyltransferase identification) as an antibody-independent approach

  • Employ CUT&RUN technology, which often provides better signal-to-noise ratio in challenging scenarios

  • Use proximity labeling approaches to identify DNA-associated proteins that interact with At5g44390

While challenging due to the protein's primary localization, properly optimized ChIP experiments could reveal unexpected nuclear functions or interactions of At5g44390 under specific conditions .

What are emerging techniques and future directions for studying At5g44390 using advanced antibody-based methods?

The study of At5g44390 can benefit from several cutting-edge antibody-based technologies and approaches:

• Advanced Imaging Applications:

  • Super-resolution microscopy: Use fluorescently-labeled At5g44390 antibodies with techniques like STORM or PALM to achieve nanometer-scale resolution of protein localization in the cell wall

  • Expansion microscopy: Physically expand plant tissues to improve resolution with standard confocal microscopy

  • Live-cell imaging: Combine nanobody technology with fluorescent proteins to track At5g44390 dynamics in living cells

  • Correlative light and electron microscopy (CLEM): Precisely localize At5g44390 at ultrastructural level

• Proximity-Based Interaction Mapping:

  • BioID or TurboID fusion proteins: Generate At5g44390 fusion constructs to biotinylate proximal proteins

  • APEX2 proximity labeling: Create At5g44390-APEX2 fusions for electron microscopy-compatible proximity labeling

  • Split-BioID systems: Investigate conditional interactions based on specific stimuli or developmental stages

  • These approaches are particularly valuable for cell wall proteins where traditional interaction methods may fail

• Single-Cell Protein Analysis:

  Technique	Application to At5g44390	Technical Considerations
  Mass cytometry (CyTOF)	Quantify At5g44390 across cell populations	Requires metal-conjugated antibody
  Single-cell Western blotting	Analyze protein heterogeneity	Microfluidic device optimization needed
  Microproteomics	Targeted protein analysis in specific cells	Limited by tissue isolation techniques
  Antibody-based FACS	Isolate cells expressing At5g44390	Protocol adaptation for plant protoplasts

• Protein Modification Analysis:

  • Develop modification-specific antibodies (phospho, glyco, etc.) for At5g44390

  • Implement multiplexed detection systems to simultaneously quantify different modified forms

  • Apply targeted mass spectrometry approaches to identify and quantify post-translational modifications

  • Map modification sites to functional domains to understand regulatory mechanisms

• Therapeutic and Biotechnological Applications:

  • Engineer antibody fragments (nanobodies, scFvs) targeting At5g44390 for in vivo modulation

  • Develop antibody-based biosensors for real-time monitoring of At5g44390 activity

  • Create immunomodulatory tools to alter At5g44390 function in specific tissues

  • Explore applications in plant biotechnology and crop improvement

• Machine Learning Integration:

  • Implement deep learning algorithms to analyze complex immunohistochemistry patterns

  • Develop predictive models for protein-protein interactions based on antibody-derived data

  • Use artificial intelligence to optimize antibody-based detection protocols

These emerging technologies promise to overcome current limitations in studying cell wall-localized proteins like At5g44390 and may reveal unexpected functions and regulatory mechanisms not accessible with conventional approaches .