Recombinant Mouse CD30

Recombinant mouse CD30 serves as a valuable research tool for investigating CD30 biology and immune regulation across multiple experimental contexts.

Key Research Applications

Immunological Studies

Recombinant mouse CD30 enables investigation of CD30/CD30L interactions in immune responses. This protein is particularly useful for studying T-cell co-stimulation and understanding how CD30 signaling influences T-cell activation and differentiation. The protein can be employed to examine CD30's role in thymic negative selection and its effects on T-cell development.

B-Cell Research

In B-cell studies, recombinant mouse CD30 facilitates examination of CD30 ligation effects on cellular proliferation and antibody production. The protein also allows researchers to investigate CD30-mediated expression of chemokine receptors and ligands such as CXCR4, CCL3, and CCL5 in B cells. This is particularly relevant for understanding germinal center dynamics and B-cell activation pathways.

Disease Model Development

Recombinant mouse CD30 is instrumental in autoimmune disease research. Studies utilizing this protein have demonstrated its critical role in type 1 diabetes pathophysiology, where CD30/CD30L interactions contribute to both the induction and effector phases of disease development. The protein enables functional blocking studies and mechanistic investigations of autoimmune processes.

Lymphoma Research

The protein serves as a tool for studying CD30-positive lymphomas, including Hodgkin lymphoma. Recombinant mouse CD30 can be used to develop and validate therapeutic approaches, including chimeric antigen receptor (CAR) T-cell constructs and monoclonal antibody strategies targeting CD30-expressing malignancies.

In Vitro Assay Development

Recombinant mouse CD30 functions as a positive regulator of apoptosis and can be incorporated into assays examining cell proliferation, differentiation, and survival pathways. The protein is suitable for cell culture experiments investigating CD30-mediated signaling cascades.

Yes, recombinant Mouse CD30 protein can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and the antibodies used. Recombinant standards are commonly employed to generate standard curves for quantitative ELISA, allowing accurate determination of CD30 concentrations in biological samples.

Key considerations and best practices:

• Assay Compatibility: Ensure that your ELISA kit or custom assay is validated to recognize both natural and recombinant Mouse CD30. Most commercial kits specify that their antibodies detect both forms, enabling the use of recombinant protein as a standard.
• Standard Preparation: Prepare the recombinant Mouse CD30 standard according to the protocol, typically by reconstituting the lyophilized protein in the provided diluent and performing serial dilutions to cover the assay’s dynamic range (e.g., 15.6–1000 pg/mL).
• Matrix Effects: For highest accuracy, dilute the recombinant standard in the same buffer or matrix as your samples (e.g., serum, plasma, or cell culture supernatant) to minimize matrix effects and ensure comparable binding and detection.
• Validation: It is recommended to run a pre-test to confirm that the recombinant standard yields a linear, reproducible standard curve and that sample quantification is accurate within the assay’s sensitivity and specificity limits.
• Documentation: Always refer to your ELISA kit’s user manual for specific instructions regarding standard preparation and compatibility. Some kits may provide their own recombinant Mouse CD30 standard, which is optimized for use with their antibodies.

Summary Table: Use of Recombinant Mouse CD30 as ELISA Standard

In summary: Recombinant Mouse CD30 is suitable as a standard for ELISA quantification if it is recognized by your assay antibodies and prepared according to validated protocols. Always verify compatibility and performance with your specific assay system.

Recombinant Mouse CD30 has been validated in published research primarily for applications in immunotherapy studies, especially in the context of chimeric antigen receptor (CAR) T cell development and functional assays targeting CD30-positive malignancies. Key validated applications include:

• Generation and validation of CAR-T cells: Recombinant Mouse CD30 is used as an antigen to engineer and test CAR-T cells targeting CD30, particularly for preclinical models of Hodgkin lymphoma and peripheral T-cell lymphoma. These studies involve both in vitro cytotoxicity assays and in vivo efficacy testing in mouse xenograft models.

• Functional assays: It serves as a target protein in in vitro cytotoxicity assays to assess the specificity and effectiveness of CD30-directed CAR-T cells or monoclonal antibodies. This includes measuring tumor cell killing and evaluating immune cell activation.

• Protein interaction studies: Recombinant Mouse CD30 is used to study ligand-receptor interactions, such as binding assays with CD30 ligand (CD30L/TNFSF8), and to investigate downstream signaling pathways relevant to apoptosis and immune regulation.

• ELISA and standardization: Recombinant Mouse CD30 is recommended as a standard in ELISA assays for quantifying CD30 or its ligands in biological samples.

• SDS-PAGE and protein characterization: It is validated for use in SDS-PAGE to confirm purity and molecular weight, supporting its use in biochemical and structural studies.

Summary Table of Validated Applications

These applications are supported by peer-reviewed studies focusing on immunotherapy, lymphoma research, and protein biochemistry. If you require protocol details or specific assay conditions, please specify the intended application.

To reconstitute and prepare Recombinant Mouse CD30 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer—typically sterile PBS or sterile distilled water—to a recommended stock concentration, commonly 100 μg/mL. Use gentle mixing and avoid vigorous agitation to prevent protein denaturation.

Step-by-step protocol:

• Equilibrate the vial to room temperature before opening to minimize condensation.
• Add sterile PBS or distilled water directly to the vial to achieve the desired concentration (e.g., 100 μg/mL is standard for many recombinant proteins).
• Gently swirl or invert the vial to mix. Do not vortex or shake vigorously, as this can cause foaming and loss of activity.
• Allow the protein to fully dissolve for 15–30 minutes at room temperature with gentle agitation.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can reduce activity and stability.
• Store aliquots at 2–8 °C for up to one week, or at –20 °C to –70 °C for longer-term storage.

Additional considerations:

• Use only sterile, endotoxin-free buffers and tubes to prevent contamination, especially for cell culture applications.
• Avoid buffers containing surfactants (e.g., SDS) or organic solvents, as these can denature the protein and harm cells.
• If the protein is to be used for functional assays, confirm the working concentration required for your specific application (e.g., titrate in pilot experiments).
• Before use in cell culture, dilute the stock solution to the final working concentration in your cell culture medium immediately prior to use.

Summary Table: Recombinant Mouse CD30 Protein Reconstitution

Always consult the specific product datasheet for any manufacturer-specific recommendations regarding buffer, concentration, and storage, as protein tags or formulations may require minor adjustments.