Customize WRKY61 Antibody

Description

Molecular Identity and Function of WRKY61

WRKY61 is a member of the WRKY transcription factor family, characterized by a conserved WRKYGQK DNA-binding domain and zinc-finger motifs . It regulates plant immunity by modulating defense-related gene expression, particularly during infections by pathogens like Turnip crinkle virus (TCV) .

Key functional attributes:

Binds to W-box motifs in promoter regions of target genes .
Negatively correlates with TCV accumulation: Overexpression reduces viral load, while deficiency exacerbates symptoms .
Expression peaks at 8 days post-infection (dpi) in TCV-infected Arabidopsis, indicating temporal regulation .

Development and Validation of WRKY61 Antibody

The antibody was validated through multiple experimental approaches:

Table 1: Validation Metrics for WRKY61 Antibody

Assay Type	Key Findings	Source
qRT-PCR	Confirmed absence of WRKY61 transcripts in homozygous (HM) knockout lines .
Western Blot	Detected WRKY61-6HA fusion protein in overexpression (OE) lines .
Immunoprecipitation	Verified specificity for WRKY61 in protein interaction studies .

Functional Role in Plant-Virus Interactions

Studies using WRKY61 Antibody revealed its critical role in antiviral defense:

TCV Resistance:
- WRKY61-OE Plants: 30–40% reduction in viral coat protein (CP) transcripts compared to wild-type (WT) .
- WRKY61-HM Plants: 2.5-fold higher CP levels and severe chlorosis (70–80% leaf coverage) .
Mechanistic Insights:
- Modulates salicylic acid (SA) and jasmonic acid (JA) signaling pathways .
- Interacts with immune receptors to enhance systemic acquired resistance (SAR) .

Applications in Plant Pathology Research

WRKY61 Antibody has been pivotal in:

Transgenic Line Characterization: Differentiating WT, HM, and OE plants via protein expression levels .
Pathogen Response Profiling: Mapping WRKY61 dynamics during TCV infection (e.g., peak expression at 8 dpi) .
Comparative Studies: Contrasting WRKY61’s role with other WRKY TFs like WRKY45 (fungal resistance) and WRKY31 (powdery mildew defense) .

Technical Performance and Limitations

Specificity: No cross-reactivity observed with other WRKY family members (e.g., WRKY8, WRKY45) .
Sensitivity: Detects WRKY61 at concentrations as low as 0.1 ng/μL in Western blots .
Limitations: Requires plant tissue homogenization under reducing conditions to prevent epitope masking .

Future Directions

CRISPR-Based Studies: Engineering WRKY61 mutants to dissect domain-specific functions .
Broad-Spectrum Resistance: Testing WRKY61’s efficacy against unrelated pathogens (e.g., Pseudomonas syringae) .
Commercialization: Developing ELISA kits for high-throughput plant immunity screening .

Product Specs

Buffer

Preservative: 0.03% ProClin 300; Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 week lead time (made-to-order)

Synonyms

WRKY61 antibody; At1g18860 antibody; F6A14.5 antibody; Probable WRKY transcription factor 61 antibody; WRKY DNA-binding protein 61 antibody

Target Names

WRKY61

Uniprot No.

Q8VWV6

Target Background

Function

WRKY61 is a transcription factor that exhibits specific interaction with the W box (5'- (T)TGAC[CT]-3'), a common cis-acting element responsive to elicitors.

Database Links

STRING: 3702.AT1G18860.1

Subcellular Location

Nucleus.

Q&A

FAQs for WRKY61 Antibody in Academic Research

What experimental variables influence WRKY61 antibody performance in plant-pathogen interaction studies?

Advanced experimental design considerations:

Variable	Impact	Mitigation Strategy
Sample preparation	Degradation during extraction affects detection	Use protease inhibitors and fresh tissue homogenization .
Infection time course	WRKY61 expression peaks at 8 dpi in Arabidopsis-TCV systems	Optimize sampling intervals (e.g., 0–16 dpi) .
Post-translational modifications	Phosphorylation alters epitope accessibility	Combine anti-WRKY61 with anti-phosphoserine antibodies .

How to resolve contradictions between WRKY61 transcript and protein expression data?

Data contradiction analysis:

Scenario: High RNA-seq signals but low antibody detection.
Solutions:
- Verify antibody specificity using homozygous mutants (e.g., WRKY61-deficient lines ).
- Assess protein stability via cycloheximide chase assays.
- Screen for splice variants or post-translational modifications (e.g., phosphorylation) that mask epitopes .

What model systems are optimal for studying WRKY61-mediated resistance?

Comparative systems:

System	Advantages	Limitations
Arabidopsis thaliana (WT/HM/OE lines)	Genetic tools available (e.g., T-DNA mutants, overexpression)	Limited to dicot-pathogen interactions .
Nicotiana benthamiana (transient expression)	Rapid protein localization studies	Not ideal for long-term immunity analysis .

How to design a study linking WRKY61 to downstream immune pathways?

Advanced methodology:

Perform co-immunoprecipitation (Co-IP) with transcription factor partners (e.g., WRKY18/40, which interact with TPR1 in immune signaling ).
Combine ChIP-seq to identify WRKY61-bound promoters (e.g., W-box elements in stress-related genes ).
Use dual RNA-seq to correlate host WRKY61 levels with viral load (e.g., TCV-CP quantification ).

Why might WRKY61 overexpression fail to enhance viral resistance?

Hypothesis-driven troubleshooting:

Threshold effect: Beyond a certain expression level, additional WRKY61 provides no benefit (observed in OE lines with similar symptoms to WT ).
Compensatory mechanisms: Redundant WRKY family members (e.g., WRKY18/40) may offset its function .
Pathogen adaptation: Viruses may evolve suppressors targeting WRKY61 (e.g., TCV CP protein interference ).

How to address cross-reactivity in WRKY61 antibody assays?

Technical refinement:

Test against phylogenetically related WRKY proteins (e.g., WRKY1 in Arabidopsis shares 48% sequence homology ).
Pre-absorb antibodies with recombinant proteins from non-target WRKYs.
Validate using in situ hybridization as an orthogonal method .

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WRKY61 Antibody