cysl-3 Antibody
Description
Biological Context of CysLT3 and Associated Antibodies
Cysteinyl leukotrienes (CysLTs)—including LTC4, LTD4, and LTE4—are lipid mediators of inflammation produced by leukocytes. They bind to G protein-coupled receptors (GPCRs):
Antibodies targeting these receptors, including hypothetical "cysl-3 Antibodies," aim to modulate inflammatory responses in diseases like asthma, allergic rhinitis, and inflammatory bowel disease (IBD) .
Pan-Specific Anti-CysLT Monoclonal Antibodies
A 2017 study generated murine monoclonal antibodies (mAbs) against CysLTs, including clone 10G4, which showed pan-specific binding:
| Antibody | Affinity (K<sub>D</sub>) | Specificity |
|---|---|---|
| 10G4 | 0.3 nM (LTC4) | Binds LTC4 > LTD4 > LTE4 |
| 2G9 | 1.1 nM (LTC4) | Prefers LTC4 |
| Hu10G4 | Retains parental affinity | Humanized variant of 10G4 |
These antibodies inhibit CysLT-mediated calcium signaling and vascular leakage in preclinical models .
Inflammatory Bowel Disease (IBD)
Anti-CysLT antibodies may stratify IBD subtypes:
| Study | UC Sensitivity | UC Specificity | CD Sensitivity | CD Specificity |
|---|---|---|---|---|
| Laass 2022 | 40.5% | 89.3% | - | - |
| Xu 2022 | 41.7% | 98.9% | - | - |
| Mahler 2017 | 18.4% | 93.6% | - | - |
PR3-ANCA assays (anti-proteinase 3 antibodies) show utility in differentiating ulcerative colitis (UC) from Crohn’s disease (CD) .
Asthma and Allergic Inflammation
CysLT3R blockade reduces LTE4-induced vascular permeability and granulocyte-platelet interactions in murine models . Antibodies like Hu10G4 are candidates for suppressing eosinophilic inflammation .
Limitations and Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Cysl-3 primarily catalyzes the formation of cysteine and acetate from O-acetylserine and hydrogen sulfide. It can also catalyze the formation of cysteine and acetate from S-sulfocysteine and hydrogen sulfide, and the formation of cyanoalanine and hydrogen sulfide from either S-sulfocysteine or O-acetylserine and hydrogen cyanide.
- The role of CYSL-3 remains unclear, although it exhibits sulfhydrylase activity in vitro. PMID: 24100226
STRING: 6239.R08E5.2a
UniGene: Cel.6233
Q&A
The following FAQs address key considerations for researchers working with the cysl-3 antibody in academic settings, synthesized from peer-reviewed methodologies and experimental frameworks.
How to validate cysl-3 antibody specificity in Western blot and immunofluorescence assays?
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Methodology:
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Perform siRNA-mediated knockdown of cysl-3 in target cell lines (e.g., HEK293T) and compare protein levels via Western blot .
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Use blocking peptides to confirm antibody-antigen binding specificity .
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For immunofluorescence, co-stain with organelle-specific markers (e.g., lysotracker for lysosomal localization) and quantify colocalization using tools like ImageJ .
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| Validation Step | Key Metrics | Reference |
|---|---|---|
| Knockdown efficiency | ≥70% reduction in target protein | |
| Blocking peptide efficacy | ≥90% signal reduction | |
| Colocalization confidence | Pearson’s coefficient >0.8 |
What cellular pathways involve cysl-3, and how to design a CRISPR screen to identify interactors?
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Pathway Context:
cysl-3 is hypothesized to regulate sulfur metabolism or inflammatory signaling, analogous to cysl-1 in C. elegans and CX3CL1 in vascular remodeling . -
CRISPR Screen Design:
How to resolve contradictory data on cysl-3 expression in atherosclerosis models?
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Contradiction Analysis:
Optimizing cysl-3 antibody for cryo-EM structural studies
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Methodological Pipeline:
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Epitope Engineering: Use RosettaAntibody to model the Fv region and optimize CDRH3 loops for antigen binding .
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Affinity Maturation: Apply computational alanine scanning to identify critical residues for binding energy .
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Validation: Test Fab fragments in surface plasmon resonance (SPR) for kinetic parameters (KD < 10 nM) .
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| Parameter | Target Specification |
|---|---|
| CDRH3 loop flexibility | RMSD < 1.5 Å (vs. model) |
| Binding affinity (KD) | ≤5 nM |
| Thermostability | Tm > 65°C |
Addressing cross-reactivity with cysl homologs (e.g., cysl-1, cysl-4)
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Strategies:
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BLAST Analysis: Identify homologous regions in cysl family proteins and exclude them during epitope selection .
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Epitope Mapping: Use peptide arrays to define minimal binding regions (15–20 aa) .
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Mutagenesis: Introduce point mutations in cross-reactive residues (e.g., D35A, K79R) and test binding via ELISA .
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Correlating cysl-3 expression with metabolomic profiles in inflammatory disease models
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Workflow:
