CYP710A3 Antibody
Description
Context of CYP710 Enzymes
The CYP710 family belongs to the cytochrome P450 superfamily, which is involved in oxidative metabolism of sterols, hormones, and xenobiotics. For example:
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CYP710C1: A plant-like sterol C-22 desaturase in Leishmania donovani catalyzes stigmasterol biosynthesis and influences drug resistance (e.g., amphotericin B) .
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CYP7A1: A cholesterol 7-alpha-hydroxylase critical for bile acid synthesis .
While CYP710A3 is not mentioned in the provided sources, its nomenclature suggests it is part of the CYP710 subfamily, which may share functional or structural similarities with other sterol-modifying enzymes.
Antibody Characterization Challenges
The lack of data on CYP710A3 highlights broader issues in antibody validation:
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Reproducibility: Studies show that ~50% of commercial antibodies fail specificity tests, even for well-characterized targets like CYP3A4 .
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Validation Methods: Knockout (KO) cell lines and orthogonal assays (e.g., Western blot, immunofluorescence) are critical for confirming antibody specificity .
Related Cytochrome P450 Antibodies
While CYP710A3 data is absent, insights from antibodies targeting other P450 enzymes may inform research strategies:
Recommendations for Researchers
If investigating CYP710A3:
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Verify Target Specificity: Use KO models or siRNA knockdown to confirm antibody binding.
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Explore Homology: Compare CYP710A3 sequences with characterized isoforms (e.g., CYP710C1) to infer potential epitopes or functional domains.
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Collaborate with Repositories: Leverage databases like the Human Protein Atlas or Antibodypedia for validation data .
Limitations and Gaps
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Data Availability: No peer-reviewed studies or commercial products referencing CYP710A3 were found in the provided materials.
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Nomenclature Check: Confirm the correct designation of the enzyme (e.g., potential typographical errors, such as CYP710A3 vs. CYP710C1).
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
KEGG: ath:AT2G28850
STRING: 3702.AT2G28850.1
Q&A
Given the lack of specific information on "CYP710A3 Antibody" in the search results, I will provide a general framework for FAQs related to antibodies in academic research, focusing on a hypothetical antibody like CYP710A3. This framework can be adapted to any specific antibody once more detailed information becomes available.
Answer:
Validating antibody specificity involves several steps:
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Western Blotting: Use the antibody to detect the target protein in cell lysates or tissue extracts. Ensure the antibody recognizes a single band of the expected molecular weight.
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Immunohistochemistry (IHC): Perform IHC on tissue sections to confirm the antibody's ability to specifically stain the target protein in its native environment.
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Knockdown/Knockout Experiments: Use siRNA or CRISPR-Cas9 to reduce or eliminate the target protein expression. If the antibody signal decreases or disappears, it supports specificity.
Answer:
Discrepancies can arise from differences in antibody specificity, epitope recognition, or experimental conditions. To resolve these:
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Use Multiple Antibodies: Validate findings with multiple antibodies targeting different epitopes of the protein.
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Optimize Experimental Conditions: Ensure consistent conditions across experiments, including buffer composition and incubation times.
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Control Experiments: Include positive and negative controls to verify antibody performance.
Answer:
Epitope mapping is crucial for understanding antibody specificity and can be achieved through:
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Peptide Array: Synthesize overlapping peptides covering the protein sequence and assess antibody binding.
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Mutagenesis: Introduce point mutations in the protein and evaluate changes in antibody binding.
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X-ray Crystallography or NMR: Determine the three-dimensional structure of the antibody-protein complex to identify the epitope.
Answer:
Cross-reactivity can be assessed and minimized by:
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Pre-adsorption: Pre-incubate the antibody with lysates from cells lacking the target protein to remove non-specific binding.
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Competitive ELISA: Use a known antigen to compete with the sample for antibody binding, reducing non-specific interactions.
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Western Blotting with Controls: Include controls with known amounts of the target protein and unrelated proteins to evaluate specificity.
Answer:
To optimize antibody performance:
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Single-Chain Variable Fragment (scFv) Engineering: Engineer smaller antibody fragments for improved penetration and specificity.
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Antibody Conjugation: Conjugate antibodies with fluorescent dyes or enzymes to enhance detection sensitivity.
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Super-resolution Microscopy: Use techniques like STORM or STED to achieve higher resolution imaging with optimized antibodies.
Answer:
Statistical analysis should include:
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Replication: Perform experiments in triplicate to ensure reproducibility.
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Normalization: Normalize data to internal controls to account for variability.
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Hypothesis Testing: Use appropriate statistical tests (e.g., t-test, ANOVA) to compare groups and determine significance.
Example Data Table for Antibody Validation
| Antibody | Western Blot | IHC | Knockdown/Knockout |
|---|---|---|---|
| CYP710A3 | Specific band at 50 kDa | Positive staining in target tissue | Signal reduction upon knockdown |
| Control | No band detected | No staining | No change upon knockdown |
This table illustrates how different validation methods can be used to confirm antibody specificity.
