CTL1 Antibody
Description
CTL1 Protein Structure and Function
CTL1 is a 70–73 kDa protein with nine transmembrane (TM) domains, featuring an intracellular N-terminus and extracellular C-terminus . Key functional insights:
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Transport Mechanism: Acts as a high-affinity choline/H⁺ and ethanolamine/H⁺ antiporter, regulating phospholipid synthesis via the Kennedy pathway .
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Conserved Domains: TM8 and TM9 are highly conserved across species and likely harbor substrate-binding sites .
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Pathological Roles: Deficiency causes neurodegeneration (e.g., leukoencephalopathy, cerebellar atrophy) due to impaired choline transport and membrane lipid depletion .
3.1. Immunodetection
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Flow Cytometry: Anti-CTL1 (ACT-021) detects surface CTL1 in live human THP-1 and Jurkat cells .
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Western Blot: Antibodies like ab110767 identify CTL1 in lysates (e.g., K562 leukemia cells) .
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Blocking Studies: Pre-absorption with blocking peptides (e.g., BLP-CT021) confirms antibody specificity .
3.2. Functional Studies
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Cancer Research: CTL1 is upregulated in malignancies (e.g., breast adenocarcinoma, colon cancer). Inhibition with Amb544925, a plant-derived CTL1 blocker, reduces tumor growth and metastasis .
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Metabolic Pathways: CTL1 mediates ethanolamine uptake for phosphatidylethanolamine synthesis, balancing CDP-choline and CDP-ethanolamine pathways .
Key Research Findings
Clinical and Therapeutic Implications
Product Specs
Target Background
CTL1 antibody targets a protein crucial for normal plant growth and development. Its functions include regulating cell expansion and differentiation, particularly in roots and hypocotyls. CTL1 prevents lignin accumulation in the pith and may modulate ethylene-mediated developmental regulation. Evidence suggests a role in establishing thermotolerance acclimation and in controlled anisotropic cell expansion, influencing root gravitropism and thigmotropism. Furthermore, CTL1 contributes to root system architecture adaptation to varying environmental conditions, such as nitrate availability, and enhances salt and potential drought tolerance by preventing excessive sodium ion accumulation. Importantly, CTL1 exhibits no chitinase activity.
Key Research Findings on CTL1 Function:
- Studies indicate a role for CTL1, a giant cell-repressed gene. (PMID: 29389847)
- Consistent with its role in sieve area development, AtCTL1 displays sustained, polar localization in developing sieve plates. (PMID: 25008948)
- CTL1 influences primary wall cellulose deposition and, along with CTL2, affects cellulose assembly and glucan-based polysaccharide cross-linking. (PMID: 22327741)
- CTL1 expression is primarily observed in roots and root tips during seedling development, with protein localization to the cell wall. (PMID: 21949212)
- AtCTL1 plays a constitutive role in cell wall synthesis and elongation, with its function modulated by environmental stimuli such as high nitrate concentrations, impacting organ morphology. (PMID: 20007445)
Q&A
Basic Research Questions
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How to validate CTL1 antibody specificity in flow cytometry applications?
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Methodological approach:
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Perform siRNA-mediated CTL1 knockdown in target cells (e.g., THP-1 monocytes) and compare antibody signal intensity pre/post-knockdown .
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Use blocking peptides (e.g., BLP-CT021) to pre-absorb the antibody and confirm loss of signal .
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Cross-validate with alternative antibodies targeting distinct epitopes (e.g., LV-58 vs. EN-627) to confirm consistent localization patterns .
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Key validation data:
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What experimental controls are critical for CTL1 antibody-based immunoblotting?
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Essential controls:
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Common pitfalls:
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Advanced Research Questions
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How to resolve contradictions in CTL1 localization data across studies?
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Analysis framework:
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Epitope accessibility: Antibodies targeting extracellular loops (e.g., VIM15) may fail to detect intracellular pools under non-permeabilized conditions .
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Subcellular fractionation: Combine differential centrifugation with antibodies targeting organelle-specific markers (e.g., mitochondrial vs. plasma membrane CTL1) .
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Functional assays: Correlate antibody signal with choline uptake inhibition (e.g., HC3 sensitivity) to confirm physiological relevance .
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Case study:
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What strategies optimize CTL1 antibody use in multiplexed imaging studies?
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Technical recommendations:
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Pair CTL1 antibodies with fluorophores emitting >600 nm to avoid overlap with autofluorescence in fixed tissues .
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Combine extracellular epitope-targeting antibodies (e.g., ACT-021) with intracellular markers (e.g., β-actin) using sequential staining protocols .
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Validate co-localization with metabolic tracers (e.g., [³H]-choline) to confirm functional compartments .
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How to design experiments investigating CTL1’s dual role in choline and ethanolamine transport?
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Experimental design:
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Radiolabeling: Compare [¹⁴C]-choline vs. [¹⁴C]-ethanolamine uptake in CTL1-overexpressing vs. knockout cells .
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Metabolic profiling: Quantify CDP-choline and CDP-ethanolamine levels via LC-MS in antibody-treated macrophages .
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Kinetic assays: Use Michaelis-Menten analysis to determine Km values for choline/ethanolamine under varying pH .
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Key findings:
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What are best practices for using CTL1 antibodies in genetic disease models?
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Guidelines:
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Animal models: Validate antibody cross-reactivity in species-specific mutants (e.g., Slc44a1⁻/⁻ mice) .
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Human tissues: Prioritize antibodies validated in disease-relevant cell lines (e.g., Colo 205 for colorectal cancer) .
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Data interpretation: Contextualize CTL1 expression changes with functional readouts (e.g., PKC activation in choline-depleted macrophages) .
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Data Contradiction Analysis
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Why do some studies report CTL1 at 70 kDa while others detect smaller bands?
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Root causes:
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Resolution strategy:
Condition Observed MW (kDa) Interpretation Native 70 Full-length, glycosylated Deglycosylated 55 Core protein Truncated 40–50 Artifact (optimize lysis)
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