CYP19-3 Antibody
Description
CYP19A1/Aromatase Overview
Aromatase (EC 1.14.14.1) is a cytochrome P450 enzyme critical for converting androgens like testosterone and androstenedione into estrogens (estradiol and estrone). The CYP19A1 gene (15q21.2) encodes this enzyme, which is expressed in gonads, brain, adipose tissue, and placenta . Dysregulation of aromatase activity is linked to estrogen-dependent cancers, osteoporosis, and endocrine disorders .
Anti-CYP19A1 Antibody Characteristics
Commercial antibodies against CYP19A1 are typically polyclonal or monoclonal reagents validated for applications such as:
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Western blot (WB)
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Immunohistochemistry (IHC)
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Immunofluorescence (IF/ICC)
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Enzyme-linked immunosorbent assay (ELISA)
Key antibody properties:
Proteintech Anti-CYP19A1 (16554-1-AP):
LSBio Anti-CYP19A1 (LS-C335211):
| Application | Reactivity | Price (USD) |
|---|---|---|
| IHC, IP, WB | Human, mouse | $275 (20 µl) |
Functional and Clinical Research Findings
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Polymorphisms and Drug Response: SNPs in CYP19A1 (e.g., rs6493497, rs7176005) correlate with aromatase inhibitor efficacy in breast cancer. Patients with these variants show altered estrogen levels and tumor responses .
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Mechanistic Studies: Electrophoretic mobility shift assays (EMSAs) and luciferase reporter systems confirm that CYP19A1 SNPs influence transcriptional regulation and estrogen synthesis .
Technical Considerations
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Antigen Retrieval: Optimal IHC results require TE buffer (pH 9.0) or citrate buffer (pH 6.0) .
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Storage: Stable at -20°C in PBS with 0.02% sodium azide and 50% glycerol .
Research Implications
Anti-CYP19A1 antibodies are pivotal for studying:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Roc1 and Roc2 interact with the Tomato bushy stunt tombusvirus RNA and replication proteins, leading to the inhibition of viral replication. PMID: 24089553
Q&A
Answer:
To validate the specificity of a CYP19A1 antibody, researchers typically use Western blot (WB), immunohistochemistry (IHC), and immunocytochemistry (ICC) with known positive and negative controls. For WB, use human placenta tissue as a positive control. For IHC, placenta tissue from humans or small intestine tissue from mice can serve as positive controls. Ensure thorough antibody incubation and use appropriate blocking agents to minimize non-specific binding .
Answer:
When encountering contradictory data, consider the following:
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Assay Conditions: Ensure that the conditions for each assay (e.g., antibody concentration, incubation time) are optimized and consistent across experiments.
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Sample Preparation: Verify that sample preparation methods are uniform and that samples are not degraded.
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Control Samples: Use consistent positive and negative controls to validate assay specificity and sensitivity.
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Statistical Analysis: Perform robust statistical analysis to account for variability and ensure that results are statistically significant.
Answer:
CYP19A1 overexpression contributes to increased estrogen production in tumors, which can affect immune responses and cancer progression . To study this, researchers can use antibodies to detect CYP19A1 expression levels in tumor samples via IHC or WB. Additionally, chromatin immunoprecipitation (ChIP) can be employed to investigate the transcriptional regulation of CYP19A1.
Answer:
Optimizing antibody concentrations is crucial for achieving specific and sensitive results. Start with recommended concentrations (e.g., 0.1-0.5 μg/mL for WB) and adjust based on signal-to-noise ratios. Use serial dilutions to find the optimal concentration that maximizes specific binding while minimizing background noise.
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To assess specificity, perform WB or IHC with lysates or tissues known to express CYP19A1 and those that do not. Use peptide blocking assays where the antibody is pre-incubated with the immunizing peptide to confirm specificity. Additionally, use mass spectrometry to validate the target protein identity in immunoprecipitation experiments.
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In situ hybridization allows for spatial localization of CYP19A1 mRNA in tissues. Use specific probes synthesized from cDNA sequences to detect mRNA expression. Real-time PCR can quantify CYP19A1 mRNA levels using primers specific to the gene. This combination provides both spatial and quantitative information on CYP19A1 expression.
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To study evolutionary conservation, use antibodies validated across multiple species. Perform WB or IHC on tissue samples from different species to compare expression patterns. Additionally, sequence alignment and phylogenetic analysis can help identify conserved regions of the CYP19A1 protein.
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Common issues include non-specific binding, low signal, or inconsistent results. Resolve these by:
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Optimizing antibody concentrations and incubation times.
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Improving sample quality and preparation methods.
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Using appropriate blocking agents and washing conditions.
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Validating antibody specificity with controls and peptide blocking assays.
Answer:
Integrate CYP19A1 antibodies with tools like ChIP-seq for transcriptional regulation studies, CRISPR-Cas9 for gene editing, and mass spectrometry for proteomic analysis. This multi-faceted approach provides a comprehensive understanding of CYP19A1's role in biological systems.
