CKX3 Antibody
Description
Introduction to CK-3 Antibody
Cytokeratin 3 (CK-3) is a type II intermediate filament protein critical for maintaining structural integrity in epithelial tissues. Monoclonal antibodies targeting CK-3, such as AE5 (ab68260) and AE1/AE3 clones, are widely used in immunohistochemistry (IHC) and Western blotting to study epithelial malignancies and cellular differentiation . These antibodies recognize the 65 kDa cytokeratin protein, encoded by the KRT3 gene, which is expressed in stratified squamous epithelia like the cornea and esophagus .
Key Properties of CK-3 Antibodies:
CK-3 antibodies exhibit high specificity for epithelial cells, enabling differentiation between carcinomas of epithelial origin and other malignancies . For example, CK-7 antibodies (related to CK-3) distinguish breast cancer cells (SKBR3) from colon cancer cells (HT29) by staining intensity .
Mechanistic Insights
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Structural Role: CK-3 forms a resilient intermediate filament network, protecting epithelial cells from mechanical stress .
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Pathway Regulation: While not a therapeutic agent itself, CK-3’s role in epithelial integrity intersects with pathways like PI3K/AKT/mTOR, which are targeted by small-molecule inhibitors (e.g., the compound CK-3 in hepatocellular carcinoma studies) .
Validation and Limitations
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Western Blot Performance: Anti-CK-3 antibody [AE5] shows clear bands at 65 kDa in transfected 293T cells, with minimal non-specific binding .
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Clinical Challenges: False-positive rates in DTC detection using CK antibodies range from 0.21 to 1.02 events per 10⁶ mononuclear cells (MNCs), necessitating rigorous controls .
Comparative Analysis of CK Antibodies
Future Directions
Emerging assays, such as cell-based platforms for autoantibody detection, could integrate CK-3 antibodies to improve diagnostics for epithelial-derived disorders . Enhanced multiplexing and reduced cross-reactivity remain priorities for clinical adoption .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Studies have shown that CKX3 and CKX5 mutants exhibit increased silque and seed formation, suggesting a regulatory role in reproductive meristem activity. PMID: 21224426
Q&A
Given the lack of specific information on "CKX3 Antibody" in the search results, I will provide a general set of FAQs that researchers might encounter when working with antibodies in academic research scenarios. These questions and answers are designed to reflect the depth of scientific research, focusing on experimental design and data analysis.
Answer:
To validate the specificity of an antibody, you should:
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Use multiple controls: Include negative controls (e.g., secondary antibody only) and positive controls (e.g., known antigen).
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Optimize conditions: Determine the optimal concentration of the primary and secondary antibodies and incubation times.
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Assess cross-reactivity: Test the antibody against closely related antigens to ensure specificity.
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Consider Western blot or ELISA: These methods can help confirm the antibody's specificity by detecting the target protein in different samples.
Answer:
Common issues in ELISA data analysis include:
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Background noise: Caused by insufficient washing, contaminated buffers, or excessive detection reagent. Address by optimizing washing steps and reagent concentrations.
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Poor standard curves: Due to errors in standard preparation or degradation. Use different scales (e.g., log-log) and ensure proper reconstitution of standards.
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Optimize assay conditions: Adjust reagent concentrations and incubation times to improve results.
Answer:
When selecting an antibody, consider:
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Specificity: Ensure the antibody is specific to your target antigen.
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Sensitivity: Choose an antibody that can detect the antigen at the expected concentration.
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Species and isotype compatibility: Ensure compatibility with your experimental system.
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Validation data: Look for antibodies with published validation data or perform your own validation experiments.
Answer:
Monoclonal antibodies can be used to study protein structure and function by:
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Identifying epitopes: Mapping epitopes can reveal structural features of proteins.
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Inhibiting enzyme activity: Antibodies can bind to specific sites on enzymes, affecting their activity.
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Detecting post-translational modifications: Antibodies can be raised against modified epitopes, allowing researchers to study protein modifications.
Answer:
Common issues in flow cytometry include:
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Non-specific binding: Use blocking agents (e.g., Fc block) to reduce non-specific binding.
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Insufficient staining: Optimize antibody concentrations and incubation times.
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Dead cell staining: Use viability dyes to exclude dead cells from analysis.
Answer:
Simple Western assays offer:
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Linear dynamic range: Detect proteins over a wide concentration range (approximately 3 orders of magnitude).
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Automated processing: Results are processed automatically, providing quantitative data such as signal intensity and area under the curve.
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High reproducibility: Eliminates inconsistencies associated with traditional Western blotting.
Answer:
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Monoclonal Antibodies: Produced by hybridoma technology, recognizing a single epitope. They are homogeneous but more susceptible to epitope loss.
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Polyclonal Antibodies: Recognize multiple epitopes, offering broader reactivity but potential for cross-reactivity.
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Generation: Monoclonal antibodies involve B cell fusion with immortal cells, while polyclonal antibodies are generated by immunizing animals with antigens.
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Purification: Affinity purification can be used to enhance specificity by removing unwanted antibodies.
