CLC-F Antibody
Description
Introduction to CLCF-1 and Its Antibody
CLCF-1 (CCL22) is a cytokine with dual roles: B-cell stimulation and neurotrophic properties. It binds to CCR4 receptors, influencing immune cell migration and activation . The anti-CLCF-1 antibody is designed to neutralize CLCF-1, mitigating its pathological effects (e.g., albuminuria, inflammatory responses).
Key Features
| Feature | Description |
|---|---|
| Target | CLCF-1 (CCL22) |
| Mechanism | Binds to CLCF-1, blocking receptor interactions and downstream signaling |
| Applications | Renal disease, autoimmune disorders, B-cell-related therapies |
Antibody Structure
The anti-CLCF-1 antibody follows the canonical IgG structure:
-
Fab Region: Contains variable domains (VH, VL) for CLCF-1 binding .
-
Fc Region: Mediates immune effector functions (e.g., complement activation, Fc receptor binding) .
CLCF-1 Pathway
CLCF-1 activates the JAK/STAT pathway, leading to:
-
STAT3 Phosphorylation: Critical for cell signaling and immune responses .
-
Podocyte Activation: Alters cytoskeleton dynamics (e.g., lamellipodia formation), increasing glomerular permeability .
Renal Dysfunction
CLCF-1 infusion in mice increases urine albumin/creatinine ratio (UACR) and renal cortical pSTAT3 levels, indicating albuminuria and inflammation . The anti-CLCF-1 antibody reverses these effects:
| Parameter | CLCF-1 Alone (5 ng/mL) | CLCF-1 + Antibody (50 µg/mL) |
|---|---|---|
| P alb (Albumin Permeability) | 0.7–0.8 | 0.05–0.1 |
| UACR | 0.57 ± 0.46 (28-day infusion) | Baseline levels restored |
Data sourced from glomerular permeability assays and chronic infusion studies .
B-Cell Modulation
CLCF-1 enhances IgG expression in splenic B cells. Anti-CLCF-1 antibody reduces this response, as shown by flow cytometry .
Therapeutic Potential
| Model | Outcome | Source |
|---|---|---|
| Renal Injury | Reduced albuminuria, normalized pSTAT3 | |
| Autoimmune Diseases | Suppressed B-cell activation | |
| Cancer Therapy | Potential synergy with immunocytokines | (contextual) |
Limitations and Challenges
-
Specificity: Anti-CLCF-1 antibodies require precise dosing to avoid off-target effects (e.g., systemic immune suppression) .
-
Delivery: Chronic infusion protocols may necessitate sustained neutralization strategies .
Acute vs. Chronic Effects
| Parameter | Acute CLCF-1 Injection (Single Dose) | Chronic CLCF-1 Infusion (28 Days) |
|---|---|---|
| pSTAT3 (Renal Cortex) | 3.0 ± 1.8-fold increase | Persistent elevation |
| UACR | 1.62 ± 0.37-fold increase | 0.57 ± 0.46 (from baseline 0.20) |
JAK/STAT Inhibition Synergy
The JAK2 inhibitor BMS911543 synergizes with anti-CLCF-1 to block CLCF-1-induced albumin permeability .
Table 1: CLCF-1-Induced Glomerular Permeability
| CLCF-1 Concentration (ng/mL) | P alb (Albumin Permeability) | Antibody Blockade (50 µg/mL) |
|---|---|---|
| 0.05 | 0.2 | 0.05 |
| 0.5 | 0.6 | 0.1 |
| 5.0 | 0.8 | 0.1 |
Adapted from dose-response studies .
Table 2: Experimental Outcomes in Murine Models
| Condition | CLCF-1 Alone | CLCF-1 + Antibody |
|---|---|---|
| Podocyte Stress Fibers | Reduced | Restored |
| B-Cell IgG Expression | Elevated | Suppressed |
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Given the lack of specific information on "CLC-F Antibody" in the search results, I will provide a general set of FAQs for researchers working with antibodies in academic research scenarios. These FAQs will cover aspects relevant to experimental design, data analysis, and methodological considerations.
Experimental Design and Controls
-
Q: What controls should I include in my antibody-based experiments to ensure reliability?
-
A: Include both positive and negative controls. Positive controls should be samples known to express the target antigen, while negative controls should lack the antigen. This helps verify the antibody's specificity and sensitivity.
Data Interpretation and Contradictions
-
Q: How do I analyze and resolve contradictory data from different antibody-based assays?
-
A: Consider factors such as antibody batch variations, differences in sample preparation, and assay conditions. Repeat experiments with consistent conditions and validate results across multiple methods (e.g., Western blot, IF) to confirm findings.
Advanced Research Questions: Antibody Engineering
-
Q: What considerations should I have when engineering antibodies for improved effector functions?
-
A: Focus on modifying the Fc domain to enhance or reduce interactions with Fc receptors, depending on the desired immune response. Human IgG1 is often used for increasing effector functions, while mutations can be introduced to minimize interactions if needed .
Methodological Considerations: Antibody Selection
-
Q: How do I choose the most appropriate antibody for my research question?
-
A: Consider the antibody's specificity, sensitivity, and compatibility with your experimental conditions. Ensure it is validated for your specific application (e.g., IF, Western blot) and has consistent performance across different lots .
Troubleshooting Common Issues
-
Q: What are common issues encountered with antibody-based experiments, and how can they be addressed?
-
A: Common issues include nonspecific binding and poor signal-to-noise ratios. These can be addressed by optimizing fixation and permeabilization conditions, adjusting antibody concentrations, and using blocking agents to reduce background staining .
Advanced Techniques: Multiplexing and Co-staining
-
Q: How can I effectively co-stain with multiple antibodies in a single experiment?
-
A: Use antibodies with distinct fluorophores and ensure they do not cross-react. Validate each antibody individually before combining them to ensure specificity and minimal spectral overlap.
Data Analysis and Quantification
-
Q: How do I quantify antibody staining in immunofluorescence experiments?
-
A: Use image analysis software to measure fluorescence intensity. Ensure consistent imaging conditions across samples, and consider using internal controls for normalization.
Future Directions: Antibody Development
-
Q: What are emerging trends in antibody development for research applications?
-
A: Trends include the use of single-domain antibodies, nanobodies, and engineered antibodies with improved stability and specificity. These advancements aim to enhance detection sensitivity and reduce background noise in various assays.
Example Data Table: Antibody Validation
| Antibody | Specificity | Sensitivity | Application |
|---|---|---|---|
| CST-123 | High | Excellent | IF, WB |
| CST-456 | Moderate | Good | WB |
| CST-789 | Low | Poor | Not Recommended |
Note: This table is a hypothetical example. In real scenarios, specific data would be based on experimental validation results.
