Recombinant Mouse CD30 Ligand

Recombinant Mouse CD30 Ligand (CD30L/TNFSF8) is a valuable tool for research applications focused on immunology, oncology, and autoimmune disease due to its specific biological functions and signaling properties.

Key scientific reasons to use recombinant mouse CD30 ligand:

• Study of T cell activation and costimulation: CD30L is a member of the TNF superfamily and acts as a ligand for CD30 (TNFRSF8), a receptor expressed on activated T cells and certain lymphoma cells. The CD30/CD30L interaction is known to induce T cell proliferation, activation, differentiation, and apoptosis, making it essential for dissecting T cell signaling pathways and immune regulation.

• Modeling immune responses and humoral immunity: CD30/CD30L interactions play a role in secondary humoral immune responses and can directly affect B cell proliferation and differentiation. Recombinant CD30L enables controlled in vitro and in vivo studies of these processes, including the modulation of antibody production and B cell function.

• Autoimmune disease research: Blocking CD30L in mouse models has been shown to inhibit the development of autoimmune diabetes (type 1 diabetes) by disrupting the induction and effector phases of autoreactive T cell expansion. Recombinant CD30L can be used to probe these mechanisms or as a control in antibody-blocking experiments.

• Cancer biology and therapeutic target validation: CD30L is expressed on Hodgkin lymphoma and other hematologic malignancies. Its interaction with CD30 is implicated in tumor cell proliferation and apoptosis. Recombinant CD30L is used to stimulate or inhibit CD30+ cell lines, model disease mechanisms, and validate therapeutic strategies targeting the CD30/CD30L axis.

• Th17 cell differentiation and inflammatory disease: CD30L/CD30 signaling is critical for Th17 cell differentiation, which is relevant for studying inflammatory and autoimmune conditions. Recombinant CD30L allows for mechanistic studies of T helper cell subset development.

• Assay development and standardization: Recombinant CD30L is suitable for use as a standard in ELISA, bioassays, and Western blotting to quantify CD30L levels or activity in biological samples.

Best practices for use:

• Employ recombinant mouse CD30 ligand in in vitro cell culture systems to stimulate or block CD30 signaling.
• Use as a control or standard in immunoassays (ELISA, Western blot).
• Apply in in vivo mouse models to study immune modulation, disease progression, or therapeutic intervention.

Summary of scientific applications:

• Immunology: T cell and B cell signaling, costimulation, differentiation.
• Oncology: Lymphoma cell biology, apoptosis, proliferation.
• Autoimmunity: Diabetes models, T cell-mediated disease.
• Inflammation: Th17 differentiation, cytokine signaling.
• Assay development: ELISA, bioassay, Western blot standards.

Recombinant mouse CD30 ligand provides a controlled, reproducible reagent for dissecting the molecular and cellular mechanisms underlying CD30/CD30L biology in mouse models and cell-based systems.

Yes, recombinant mouse CD30 ligand (TNFSF8) can be used as a standard for quantification or calibration in ELISA assays, provided it is suitable for your specific ELISA format and detection system.

Key Points:

• Recombinant Protein as Standard: Recombinant proteins, such as recombinant mouse CD30 ligand, are commonly used to generate standard curves in ELISA assays. They provide a known concentration of the target analyte, allowing for accurate quantification of unknown samples. (See reference for general guidelines on ELISA standard preparation.)

• Purity and Characterization: For reliable results, the recombinant protein should be highly purified and its concentration accurately determined (e.g., by amino acid analysis, HPLC, or spectrophotometry). The protein should also be free of contaminants that could interfere with the assay.

• Compatibility with ELISA Kit: If you are using a commercial ELISA kit, check the manufacturer’s instructions to confirm whether their kit is compatible with recombinant mouse CD30 ligand as a standard. Some kits may specify the use of a particular standard or may have been validated with a specific recombinant protein. (See references , , , , , , for examples of ELISA kits that use recombinant standards.)

• Standard Curve Preparation: Prepare a dilution series of the recombinant protein in the appropriate buffer (often the same as the sample diluent) to create a standard curve. The range of the standard curve should cover the expected concentration of CD30 ligand in your samples. (See reference for guidelines on standard curve setup.)

• Validation: It is important to validate that the recombinant protein behaves similarly to the native protein in your assay system. This includes confirming that the antibody used in the ELISA recognizes both the recombinant and native forms of CD30 ligand.

Summary:

Recombinant mouse CD30 ligand can be used as a standard for ELISA quantification, but ensure it is pure, accurately quantified, and compatible with your ELISA system. Always follow the manufacturer’s recommendations if using a commercial kit, and validate the standard curve for your specific application.

Recombinant Mouse CD30 Ligand (CD30L/TNFSF8) has been validated for multiple applications in published research, including bioassays, ELISA development, in vivo studies, and Western blotting. Additional studies have demonstrated its use in T cell co-stimulation, mitogenic assays, and functional studies related to lymphoma and autoimmune disease models.

Key validated applications:

• Bioassays: Used to assess biological activity, such as co-stimulation of T cells and mitogenic effects on Hodgkin's disease-derived cell lines.
• ELISA Development: Serves as a standard or capture antigen for quantifying CD30L/TNFSF8 levels or activity in immunoassays.
• In Vivo Studies: Applied in murine models to investigate the role of CD30/CD30L interactions in disease, including autoimmune diabetes and lymphoma.
• Western Blot: Used as a positive control or antigen for antibody validation and detection of CD30L protein expression.
• Functional Immunology: Employed to study T cell activation, class switching, and antibody production in B cells, as well as disruption of CD30-CD30L signaling in lymphoma cell survival.
• Therapeutic Target Validation: Utilized in research targeting CD30L for therapeutic intervention in B-cell non-Hodgkin lymphoma and autoimmune diseases.

Additional relevant details:

• Cell Culture: Recombinant CD30L is recommended for use in cell or tissue culture experiments, particularly when formulated with BSA for stability.
• Mechanistic Studies: Used to dissect the molecular interactions between CD30 and CD30L in immune regulation and disease pathogenesis.
• Antibody Validation: Recombinant CD30L is used as an antigen for validating anti-CD30L antibodies in flow cytometry and immunofluorescence assays.

These applications are supported by peer-reviewed publications and product validation data, demonstrating the versatility of recombinant mouse CD30 ligand in immunological, oncological, and autoimmune research contexts.

To reconstitute and prepare Recombinant Mouse CD30 Ligand protein for cell culture experiments, centrifuge the vial briefly before opening, then reconstitute the protein in sterile distilled water or sterile PBS to a final concentration of 0.1–0.5 mg/mL. Avoid vigorous mixing such as vortexing or harsh pipetting to prevent protein denaturation.

Step-by-step protocol:

• Centrifuge the vial to collect the lyophilized protein at the bottom before opening.
• Add sterile distilled water or PBS to achieve a concentration between 0.1–0.5 mg/mL (commonly 0.5 mg/mL in PBS is used for cell culture applications).
• Gently mix by swirling or inverting the vial; do not vortex or pipette up and down vigorously.
• Allow the protein to dissolve at room temperature for 5–10 minutes, gently mixing occasionally until fully dissolved.
• Aliquot the reconstituted protein to avoid repeated freeze-thaw cycles, which can degrade protein activity.
• Store aliquots at 2–8°C for up to 1 month, or at –20°C to –70°C for longer-term storage.

Preparation for cell culture:

• If needed, further dilute the stock solution in sterile cell culture medium or PBS to the desired working concentration immediately before use.
• Filter sterilize the working solution if sterility is a concern and the buffer composition allows.
• Always include appropriate controls in your cell culture experiments.

Key points:

• Use sterile technique throughout to prevent contamination.
• Avoid repeated freeze-thaw cycles by aliquoting.
• Do not vortex or pipette harshly to maintain protein integrity.

These steps will ensure the recombinant protein is properly reconstituted and ready for use in cell culture assays.