Recombinant Mouse CD40

Recombinant Mouse CD40 is a valuable tool in research because it enables precise investigation and manipulation of the CD40-CD40L signaling pathway, which is central to immune regulation, disease modeling, and therapeutic development.

Key reasons to use recombinant mouse CD40 in research applications:

• Immune System Studies: CD40 is predominantly expressed on antigen-presenting cells (APCs) such as B cells, dendritic cells, and macrophages. Its interaction with CD40L (CD154) is essential for B cell activation, antibody production, and T cell priming, making recombinant CD40 crucial for dissecting these immune processes in vitro and in vivo.
• Cancer Immunotherapy Research: CD40 signaling can activate APCs, enhance anti-tumor T cell responses, and remodel the tumor microenvironment. Recombinant mouse CD40 is used to study these mechanisms, test agonistic antibodies, and develop new immunotherapeutic strategies, including combination therapies with checkpoint inhibitors or chemotherapy.
• Autoimmunity and Inflammation: The CD40-CD40L axis is implicated in autoimmune diseases and inflammatory disorders. Recombinant CD40 allows for the modeling of these diseases and the evaluation of potential therapeutic interventions targeting this pathway.
• Neurological Disease Models: Recent research highlights the role of CD40 signaling in neuroinflammation and neurodegenerative diseases. Recombinant mouse CD40 can be used to study microglial and astrocyte activation, as well as to test inhibitors that may protect against neuronal damage.
• Drug Screening and Antibody Development: Recombinant CD40 is essential for screening and characterizing antibodies or small molecules that modulate CD40 signaling, supporting the development of targeted therapies for cancer, autoimmunity, and other diseases.
• Cellular and Molecular Mechanism Elucidation: Using recombinant proteins enables controlled studies of CD40-mediated signal transduction, receptor-ligand interactions, and downstream effects, which are difficult to isolate in complex biological systems.

Applications include:

• Bioassays for immune cell activation
• In vivo disease modeling
• Screening of agonists/antagonists
• Functional validation of antibodies
• Mechanistic studies of immune signaling pathways

In summary, recombinant mouse CD40 is a versatile reagent that supports a wide range of immunological, oncological, and neurobiological research by enabling targeted manipulation and analysis of the CD40 pathway.

Yes, recombinant Mouse CD40 protein can be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity and its concentration is accurately determined.

For ELISA quantification, the standard curve should be generated using a purified recombinant protein that matches the analyte detected by your assay. Recombinant Mouse CD40 is commonly used as a standard in commercial Mouse CD40 ELISA kits, which recognize both natural and recombinant forms of the protein. This allows for accurate quantification of CD40 in biological samples by comparison to the standard curve generated from known concentrations of the recombinant protein.

Key considerations for using recombinant Mouse CD40 as an ELISA standard:

• Purity and Formulation: The recombinant protein should be highly purified and, if possible, formulated with a carrier protein such as BSA to enhance stability and reproducibility.
• Accurate Concentration: The concentration of the recombinant standard must be precisely known, typically determined by absorbance at 280 nm or another validated method.
• Standard Curve Preparation: Prepare serial dilutions of the recombinant protein to cover the expected range of sample concentrations (e.g., 0–1000 pg/mL).
• Validation: Confirm that your ELISA antibodies recognize the recombinant form equivalently to the native protein. Most commercial kits are validated for both forms.
• Reconstitution and Handling: Follow manufacturer instructions for reconstitution and storage to maintain protein integrity and activity.

Best Practices:

• Use freshly prepared standard solutions and avoid repeated freeze-thaw cycles.
• Run standards in duplicate or triplicate for accuracy.
• Construct a new standard curve for each assay plate to account for inter-assay variability.

In summary, recombinant Mouse CD40 is suitable as an ELISA standard for quantification, provided it is properly prepared and validated for your specific assay system.

Recombinant Mouse CD40 has been validated in published research primarily for bioassays involving immune cell activation, signaling studies, and functional assays in mouse models. The most common applications include:

• Bioassays: Used to study CD40-mediated signaling, B cell activation, T cell-dependent immune responses, and dendritic cell maturation in vitro using whole mouse cells.
• In vivo studies: Applied in mouse models to investigate immune modulation, vaccine adjuvant effects, and therapeutic interventions targeting the CD40 pathway.
• Functional assays: Employed to assess proliferation, differentiation, and cytokine production of B cells, dendritic cells, and other antigen-presenting cells upon CD40 engagement.
• Cancer immunotherapy research: Utilized in preclinical models to evaluate the role of CD40 activation in anti-tumor immune responses and combination immunotherapy strategies.

Supporting details and context:

• Bioassay validation: Multiple studies have used recombinant mouse CD40 or its ligand (CD40L) to activate B cells, dendritic cells, and other immune cells in vitro, measuring outcomes such as proliferation, immunoglobulin class switching, and cytokine production.
• In vivo validation: Recombinant CD40 and agonistic antibodies have been used in mouse models to study immune activation, vaccine adjuvant effects, and anti-tumor efficacy, including tumor clearance and T cell expansion.
• Mechanistic studies: Research has validated recombinant mouse CD40 in dissecting downstream signaling pathways (e.g., AKT activation, NF-κB, MAPK8, PAK2) and the role of CD40 in hematopoiesis, inflammation, and immune regulation.
• Therapeutic evaluation: Recombinant mouse CD40 has been used to test novel immunotherapeutic approaches, such as bispecific antibodies and Fc-engineered agonists, in preclinical cancer models.

Summary of validated applications:

• Bioassays (in vitro functional studies)
• In vivo immunological studies (mouse models)
• Immune cell activation and differentiation assays
• Cancer immunotherapy research
• Mechanistic signaling studies

These applications are well-supported by published research, particularly in the context of mouse immunology and preclinical therapeutic development.

To reconstitute and prepare Recombinant Mouse CD40 protein for cell culture experiments, follow these general steps, which are consistent with best practices for recombinant protein handling and supported by multiple technical protocols:

1. Reconstitution

• Check the product datasheet for specific instructions, as formulation and recommended diluent may vary by protein preparation. Most recombinant mouse CD40 proteins are supplied lyophilized and should be reconstituted in sterile PBS (phosphate-buffered saline).
• If the protein contains carrier proteins (e.g., BSA), reconstitute at the recommended concentration (commonly 100 μg/mL) in sterile PBS containing at least 0.1% human or bovine serum albumin to stabilize the protein and prevent adsorption to surfaces.
• If the protein is carrier-free, reconstitute in sterile PBS alone.
• Gently add the appropriate volume of buffer directly to the vial. Allow the protein to dissolve for 15–30 minutes at room temperature with gentle agitation. Avoid vigorous shaking or vortexing to prevent foaming and protein denaturation.

2. Aliquoting and Storage

• After reconstitution, aliquot the protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade protein activity.
• Store aliquots at –20°C to –70°C for long-term storage, or at 2–8°C for short-term use (up to 1 month).
• Use a manual defrost freezer for long-term storage and avoid repeated freeze-thaw cycles.

3. Preparation for Cell Culture

• Before adding to cell cultures, dilute the reconstituted protein to the desired working concentration using cell culture medium or sterile PBS with 0.1% BSA, as appropriate for your assay.
• Filter-sterilize the final working solution if sterility is a concern and the protein preparation is not already sterile-filtered.
• Add the protein to cell cultures under sterile conditions.

4. Additional Notes

• If using the protein as a standard in ELISA or other assays, some protocols recommend reconstituting in sterile distilled water or a specific sample diluent provided with the kit. Always refer to the assay protocol for these cases.
• For functional assays (e.g., B cell activation), confirm the bioactivity of the reconstituted protein using a relevant bioassay if possible.

Summary Table: Key Steps for Recombinant Mouse CD40 Protein Reconstitution

Always consult the specific product datasheet for any unique requirements, as formulations and recommended procedures may vary between preparations.