SQS2 Antibody
Description
SQS2 Antibody: Definition and Context
SQS2 antibodies are immunoreagents designed to detect or inhibit squalene synthase 2. These antibodies are critical for studying SQS2’s role in sterol biosynthesis, particularly in plants like Salvia miltiorrhiza, where SQS2 contributes to secondary metabolite production (e.g., tanshinones) .
Key Features of SQS2:
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Molecular Weight: ~47.16 kDa (predicted for S. miltiorrhiza SQS2) .
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Conserved Domains: Six signature regions, including substrate-binding pockets and catalytic sites (DXXXD motifs) .
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Tissue Expression: Higher in roots than leaves, as shown via qRT-PCR .
Recombinant Protein Detection
In studies on S. miltiorrhiza SQS2, antibodies against GST tags (e.g., anti-GST) were used to validate recombinant SQS2ΔTM (truncated form) expressed in E. coli. This approach bypasses the need for SQS2-specific antibodies by leveraging tag-specific detection .
Methodology:
Outcome:
Antibody Characterization Challenges
While SQS2-specific antibodies are not explicitly described in the literature, broader antibody characterization principles apply:
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Validation: KO cell lines are gold-standard controls for antibody specificity (e.g., Western blotting, immunofluorescence) .
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Performance: Recombinant antibodies often outperform monoclonal/polyclonal variants in specificity and affinity .
Tissue-Specific Expression of SQS2
qRT-PCR analysis in S. miltiorrhiza revealed:
| Tissue | Relative Expression Level | Significance |
|---|---|---|
| Roots | Higher than leaves | Critical for terpenoid biosynthesis |
| Leaves | Baseline | Reference for comparative analysis . |
Functional Insights from SQS2 Studies
SQS2’s role in isoprenoid biosynthesis is defined by:
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Catalytic Activity: Converts FPP to squalene via a two-step mechanism (1,2-farnesyl shift → squalene formation) .
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Structural Features:
Gaps and Future Directions
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SQS2-Specific Antibodies:
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Limited data on antibodies directly targeting SQS2 epitopes.
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Potential applications: Inhibiting SQS2 activity in metabolic engineering or cancer research (squalene synthase inhibitors target oncogenic pathways).
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Cross-Species Reactivity:
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SQS2 is conserved across plants, but antibody cross-reactivity with human SQS (e.g., in therapeutic contexts) remains unexplored.
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Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Given the lack of specific information on "SQS2 Antibody" in the search results, I will create a collection of FAQs that generally apply to antibody research, focusing on experimental design, data analysis, and methodological considerations. These questions and answers are designed to reflect the depth of scientific research in the field of antibodies.
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Researchers typically use a combination of in vitro and in vivo assays to evaluate antibody specificity and affinity. This includes techniques like ELISA (Enzyme-Linked Immunosorbent Assay) for specificity and surface plasmon resonance (SPR) for affinity measurements. Additionally, phage display and yeast display systems are used to screen large libraries of antibodies for desired properties .
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For analyzing antibody sequencing data, researchers use software tools like Geneious, which allows for quality control, assembly, and annotation of sequences. This includes clustering sequences to identify patterns and trends, visualizing amino acid variability, and comparing datasets to understand antibody diversity and evolution .
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Validation involves confirming the antibody's specificity and affinity using techniques such as Western blot, immunofluorescence, and ELISA. Characterization includes determining the antibody's isotype, epitope mapping, and assessing its stability and functionality under various conditions .
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When encountering contradictory results, researchers often reassess experimental conditions, such as the choice of cell lines or viral strains used in neutralization assays. They may also consider the impact of antibody concentration, incubation times, and the presence of other factors that could influence assay outcomes .
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Emerging trends include the use of single-domain antibodies (sdAbs) for their smaller size and higher stability, and employing biophysics-informed modeling to design antibodies with customized specificity profiles. These approaches leverage computational tools to predict and optimize antibody sequences for improved therapeutic applications .
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Reproducibility is ensured by using standardized protocols, validated reagents, and consistent experimental conditions. Additionally, researchers often perform experiments in triplicate and use statistical analysis to confirm significant findings. Regularly updating and refining methods based on new evidence also helps maintain high standards of reproducibility .
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Glycoprotein profiling is crucial because the glycosylation pattern of antibodies can significantly affect their stability, efficacy, and safety. Techniques like LC-MS are used to monitor glycovariants, which can influence antibody-dependent cellular cytotoxicity (ADCC) and other immune responses .
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Antibodies should be reconstituted according to the manufacturer's instructions, typically in PBS with a stabilizing carrier protein like BSA. Storage conditions depend on the antibody type; generally, they are stored at -20°C with glycerol to prevent freeze-thaw cycles. Some antibodies may require additional preservatives to prevent microbial growth .
These FAQs cover a range of topics relevant to antibody research, from experimental design to data analysis and methodological considerations. They are designed to provide insights into the depth and complexity of scientific research in this field.
Data Table Example:
| Technique | Application | Advantages |
|---|---|---|
| ELISA | Specificity | High throughput, cost-effective |
| SPR | Affinity | Real-time kinetics, high sensitivity |
| Phage Display | Library Screening | Rapid selection of high-affinity antibodies |
Research Findings:
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Antibody Specificity and Affinity: Studies have shown that antibodies targeting specific epitopes can significantly enhance therapeutic efficacy. Techniques like phage display allow for rapid identification of high-affinity antibodies .
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Glycoprotein Profiling: Research highlights the importance of glycosylation in antibody stability and function. Advanced LC-MS techniques enable detailed glycovariant analysis, crucial for optimizing therapeutic antibodies .
