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What is Mouse Antibody Isotyping?
Antibody isotyping refers to the process of determining the class and subclass of antibodies, which are immunoglobulins (Ig), present in a sample. The identification of the antibody isotype is crucial for understanding how the immune system responds to pathogens, foreign substances, or therapeutic agents. This process is particularly significant in preclinical and clinical research, especially in studies involving animal models such as mice, which are commonly used in immunology and vaccine research.
In mice, antibodies are primarily classified into several major isotypes: IgG, IgM, IgA, IgE, and IgD, with IgG further divided into subclasses (IgG1, IgG2a, IgG2b, IgG3). Each isotype plays a specific role in the immune response and interacts differently with various components of the immune system.
Mouse Antibody Isotyping Panel at Creative Proteomics
Creative Proteomics offers a Mouse Antibody Isotyping 7-plex Panel Service that is specifically designed for precise and multiplexed identification of mouse antibody isotypes. By leveraging the Luminex xMAP Technology, we ensure that each assay delivers superior performance, high sensitivity, and reproducibility. This service is optimized for both high-throughput screening and small-scale studies, making it an ideal solution for various research applications.
Detection Method
Magnetic bead-based Luminex multiplex assay
Species
Mouse
Analytes Detected
| Species | Specification | Protein Targets | Applications | Price |
|---|---|---|---|---|
| Mouse | Mouse Antibody Isotyping 7-plex Panel | IgA, IgE, IgG1, IgG2b, IgG2c, IgG3, IgM | Suitable for analyzing immune responses in mouse models, including vaccine research, therapeutic antibody development, and immune monitoring in disease models. | +Inquiry |
Sensitivity (Detection Limits)
- IgA: 1–50 ng/mL
- IgE: 0.5–50 ng/mL
- IgG1: 0.5–10 ng/mL
- IgG2b: 0.5–10 ng/mL
- IgG2c: 0.5–10 ng/mL
- IgG3: 0.5–10 ng/mL
- IgM: 1–50 ng/mL
Advantages of Mouse Antibody Isotyping Luminex Assay
- Multiplexing Capability: The Luminex xMAP Technology allows simultaneous detection of multiple antibody isotypes in a single sample, increasing efficiency and saving valuable time and resources.
- High Sensitivity: The assay offers high sensitivity, enabling the detection of even low-abundance antibodies, ideal for early-stage immune response studies or small sample volumes.
- Quantitative Results: Provides precise quantitative data on antibody concentrations, offering a clear view of immune responses in various experimental conditions.
- Wide Detection Range: The Luminex assay can detect a broad range of antibody isotypes, including IgA, IgE, IgG1, IgG2b, IgG2c, IgG3, and IgM, making it versatile for diverse research applications.
- Scalability and Customization: Suitable for both small-scale studies and large, high-throughput applications. The assay can be tailored to meet specific research needs and antibody targets.
- Time and Cost Efficiency: By consolidating multiple isotype analyses into one panel, the Luminex assay minimizes the need for separate tests, thus reducing both time and cost.
- Reproducibility and Reliability: Luminex xMAP technology provides consistent, reproducible results, ensuring reliable data for both preliminary and advanced research.
Sample Requirements for Mouse Antibody Isotyping Luminex Panel
| Sample Type | Minimum Volume | Collection & Preparation | Storage Conditions |
|---|---|---|---|
| Serum | 50–100 µL | Store at -20°C or -80°C for long-term storage | No special preparation required; clear serum preferred |
| Plasma | 50–100 µL | Store at -20°C or -80°C for long-term storage | Separate plasma from whole blood immediately after collection |
| Cell Culture Supernatant | 50–100 µL | Store at -20°C or -80°C for long-term storage | Centrifuge to remove debris, clear supernatant preferred |
| Tissue Homogenate | 50–100 µL | Store at -20°C or -80°C for long-term storage | Homogenize tissue sample in an appropriate buffer and centrifuge to remove debris |
Application of Mouse Antibody Isotyping Panel
- Immunology and Vaccine Research
In vaccine development, knowing the specific isotype profile of antibodies induced by a vaccine is essential for evaluating immune response quality. For example, IgG2a is typically associated with Th1-type immune responses, which are important for protection against intracellular pathogens, whereas IgG1 is more related to Th2-type responses, important for extracellular pathogens. Antibody isotyping in vaccine research helps determine which isotypes are most protective and informs the design of future vaccines.
- Therapeutic Antibody Development
Monoclonal antibodies (mAbs) are widely used in therapeutics, particularly in cancer treatment and autoimmune diseases. Understanding the isotype of therapeutic antibodies is crucial, as different isotypes can have different effects on immune activation, effector functions, and pharmacokinetics. For example, IgG subclasses like IgG1 are favored in therapeutic antibodies because of their ability to activate immune cells such as macrophages and NK cells.
- Autoimmune Disease Research
Autoimmune diseases are characterized by the production of antibodies against self-antigens. Monitoring the antibody isotype profile of these antibodies can provide valuable insights into the pathogenesis of these diseases. For instance, a shift from IgM to IgG production is a hallmark of chronic autoimmune disease progression.
- Cancer Immunology
In cancer research, measuring the antibody isotype can help evaluate the immune system's response to tumors. Tumor-specific antibodies can elicit a variety of immune responses depending on the isotype, influencing the tumor's progression or regression.
- Infectious Disease Models
Mouse models are commonly used to study the immune response to infectious agents. The type of antibody response—whether dominated by IgM, IgG, or IgA—provides insights into the effectiveness of the immune system against the pathogen. Isotype switching, which is the process of changing from one antibody class to another, is an important aspect of adaptive immunity.
In addition to preconfigured panels, we also offer customized analysis services. You can customize your own panel through our customization tool, or directly email us the targets you are interested in. A professional will contact you to discuss the feasibility of customization. We look forward to working with you!
| Protein Target | Description |
|---|---|
| IgA | An antibody isotype primarily found in mucosal areas, providing protection against pathogens at surfaces like the respiratory and gastrointestinal tracts. |
| IgE | An antibody involved in allergic reactions and defense against parasitic infections; binds to allergens and triggers mast cell activation and histamine release. |
| IgG1 | The most abundant IgG subclass in mice, associated with Th2 immune responses and effective in neutralizing pathogens and activating immune cells. |
| IgG2b | An IgG subclass known for its ability to activate complement and modulate immune responses, playing a role in immune regulation. |
| IgG2c | An IgG subclass involved in Th1-type immune responses, contributing to the defense against viral and intracellular pathogens. |
| IgG3 | A subclass of IgG that plays a significant role in immune responses against viruses and parasites, particularly through complement activation. |
| IgM | The first antibody produced in response to an infection, primarily found in the bloodstream and crucial for initiating the immune response and complement activation. |
What is the accuracy of the isotyping panel results?
Our Mouse Antibody Isotyping Panel has been optimized for high sensitivity and specificity, with accuracy rates exceeding 95% for most standard isotypes. The multiplex assay format minimizes the risk of cross-reactivity, ensuring reliable classification of your samples. However, for rare or unusual antibody profiles, additional confirmatory tests may be recommended.
Can the isotyping panel differentiate between mouse IgG subclasses?
Yes, the panel can differentiate between the four major mouse IgG subclasses: IgG1, IgG2a, IgG2b, and IgG3. Identifying the subclass of an antibody is crucial because each subclass has distinct properties that can influence immune responses. For example, IgG1 and IgG2a subclasses typically have different affinities for Fc receptors, which can impact their ability to induce antibody-dependent cellular cytotoxicity (ADCC) or complement activation. The panel will provide you with a detailed breakdown of each subclass within your sample.
