BSMT1 Antibody
Description
Biological Role of BSMT1
BSMT1 catalyzes the methylation of salicylic acid (SA) to form methyl salicylate (MeSA), a volatile compound involved in:
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Systemic acquired resistance (SAR) against pathogens
Table 1: BSMT1 Functional Overview
| Property | Detail |
|---|---|
| Gene ID | At3g11480 (Arabidopsis thaliana) |
| Subcellular Localization | Cytoplasm, chloroplasts |
| Key Substrates | Salicylic acid, benzoic acid |
| Mutant Phenotype | Early senescence, altered SA accumulation |
Applications of BSMT1 Antibody in Research
BSMT1 antibody is utilized across multiple experimental paradigms to investigate protein function and localization:
Key Techniques
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Western Blot: Quantifies BSMT1 expression levels in plant tissues under stress or developmental conditions .
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Immunohistochemistry: Maps BSMT1 localization in chloroplasts and nuclei of Arabidopsis leaves .
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Chromatin Immunoprecipitation (ChIP): Identifies DNA-binding interactions, such as BSMT1’s regulatory role in SA biosynthesis genes (e.g., PAL1, ICS1) .
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Mutant Complementation Studies: Validates functional rescue in why1 mutants by detecting restored BSMT1 expression .
Table 2: Experimental Applications
Mechanistic Insights
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BSMT1 expression is suppressed in why1 mutants, leading to premature SA accumulation and accelerated senescence .
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Nuclear-localized WHIRLY1 (nWHY1) represses BSMT1 transcription, while plastid-localized WHY1 (pWHY1) enhances ICS1 expression, creating a regulatory balance .
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BSMT1 knockdown increases free SA levels by 2.5-fold in Arabidopsis, altering redox signaling .
Developmental Regulation
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BSMT1 activity peaks during late developmental stages (42 days after germination), correlating with senescence onset .
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Dual-localized WHY1 isoforms modulate BSMT1’s role in SA methylation, affecting cross-talk between jasmonic acid and SA pathways .
Validation and Specificity Considerations
BSMT1 antibody requires rigorous characterization to ensure:
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Target Specificity: No cross-reactivity with homologs like BSMT2 or BAHD-family methyltransferases .
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Assay Compatibility: Optimization for plant-specific matrices (e.g., lignocellulosic tissue) .
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Functional Correlation: Transcript-protein level consistency confirmed via parallel RNA-Seq and Western Blot .
Table 3: Validation Metrics
| Parameter | Benchmark |
|---|---|
| Epitope | C-terminal domain (aa 210–350) |
| Cross-Reactivity | None detected against Arabidopsis thaliana proteome |
| Sensitivity (Western) | Detects ≥10 ng recombinant BSMT1 |
Challenges and Future Directions
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Antibody Renewability: Recombinant BSMT1 antibodies are being developed to address batch variability .
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Structural Insights: Cryo-EM studies using BSMT1 antibody could resolve substrate-binding dynamics .
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Therapeutic Potential: Engineered plant-derived antibodies may enable novel SA-based crop protection strategies .
Product Specs
Components: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- An Arabidopsis thaliana BSMT1 knockout mutant (Atbsmt1) demonstrated an inability to accumulate methyl salicylate following pathogen infection. These plants also exhibited impaired salicylic acid and its glucoside accumulation in uninoculated leaves and failed to develop systemic acquired resistance (SAR). [AtBSMT1] PMID: 19958141
- Studies revealed that a T-DNA insertion mutation in AtBSMT1 resulted in significantly reduced methyl salicylate levels upon infection with Pseudomonas syringae. PMID: 19669626
Q&A
Based on a comprehensive analysis of available research literature, below are academically oriented FAQs addressing key methodological considerations for bispecific antibody research, though it should be noted that "BSMT1 Antibody" is not explicitly referenced in current peer-reviewed studies. The following questions and answers are generalized for bispecific antibodies (BsAbs) in oncology, synthesizing insights from multiple myeloma, solid tumor, and immunological studies.
How do bispecific antibodies differ mechanistically from monoclonal antibodies in targeting tumor microenvironments?
BsAbs engage two distinct antigens simultaneously, enabling dual-pathway modulation. For example:
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Structural advantage: BsAbs like BCMA×PDL1 IgG1 bind both tumor antigens (e.g., BCMA on myeloma cells) and immune checkpoints (e.g., PDL1 in the microenvironment), achieving localized immune activation .
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Functional synergy: Blocking PD-1/PDL1 alongside tumor-antigen targeting enhances T-cell cytotoxicity while reducing systemic toxicity compared to monoclonal antibody combinations .
Key Experimental Design Considerations:
What validation methods ensure target specificity and binding affinity in BsAb development?
Stepwise validation protocols:
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Surface plasmon resonance (SPR): Quantify binding kinetics (KD values) for both targets (e.g., BCMA×PDL1 showed nM affinity for both antigens) .
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Flow cytometry: Confirm binding to native membrane-bound antigens using primary cells (e.g., MM cells stained with anti-BCMA and anti-PDL1) .
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Functional blocking assays: Measure IC50 for antigen inhibition (e.g., nM-range blocking efficacy for PDL1 and BCMA) .
Common pitfalls: Steric hindrance may reduce affinity for one target (e.g., 10-fold lower PDL1 affinity in BCMA×PDL1 due to Fab positioning) .
How can researchers address contradictory efficacy data in BsAb trials?
Case example: Discrepancies in tumor killing (e.g., 75% MM cell death in vitro vs. limited in vivo efficacy) .
Resolution strategies:
What experimental designs optimize preclinical-to-clinical translation for BsAbs?
Recommended frameworks:
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Multiple-baseline/multiple-probe designs: Assess dose-response across 3+ cell lines or animal models to establish reproducibility .
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Withdrawal (ABAB) designs: Evaluate reversibility of BsAb effects on tumor growth and immune parameters .
Critical parameters:
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Target saturation thresholds (e.g., ≥80% BCMA occupancy required for MM cell lysis) .
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Immune cell ratios (e.g., effector:target ≥ 5:1 in PBMC co-cultures) .
How do bispecific antibodies overcome primary resistance mechanisms in hematologic malignancies?
Mechanistic insights:
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TME reprogramming: BCMA×PDL1 BsAb reduces PD-1+ T-cell exhaustion while enhancing NK-mediated ADCC .
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Dual checkpoint inhibition: c-MET/PD-1 BsAb blocks both HGF-driven metastasis and PD-1 immunosuppression .
Validation workflow:
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Ex vivo cytotoxicity assays: 7-day co-cultures with patient-derived PBMCs .
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Single-cell RNA sequencing: Identify transcriptional shifts in T-cell subsets post-treatment .
