sCD40L Human
Soluble CD-40 Ligand/TRAP Human Recombinant
Cat. No.
BT91
Source
Escherichia Coli.
Synonyms
CD40-L, Tumor necrosis factor ligand superfamily member 5, TNF-related activation protein, TRAP, T cell antigen Gp39, CD154 antigen, sCD40, IGM, IMD3, HIGM1, T-BAM, TNFSF5, hCD40L.
Appearance
Sterile Filtered White lyophilized (freeze-dried) powder.
Purity
Greater than 97.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description
sCD40L Human Recombinant produced in E.Coli is a non-glycosylated, Polypeptide chain containing 149 amino acids and having a molecular mass of 16308 Dalton.
The sCD40L is purified by proprietary chromatographic techniques.
Product Specs
Introduction
CD40L, also known as CD154, is a membrane glycoprotein found on the surface of T cells. It plays a critical role in the immune response by activating B cells and other immune cells. When CD40L binds to its receptor, CD40, on B cells, it triggers B cell proliferation, differentiation, and antibody production. CD40L also stimulates the production of cytokines, which are signaling molecules that regulate immune responses. Additionally, CD40L has been shown to activate monocytes, enhancing their ability to kill tumor cells. It can also costimulate the proliferation of activated T cells, leading to the production of cytokines such as IFN-gamma, TNF-alpha, and IL-2.
Description
Recombinant human sCD40L, produced in E. coli, is a non-glycosylated polypeptide chain comprising 149 amino acids with a molecular weight of 16308 Daltons. This protein is purified using proprietary chromatographic techniques to ensure high purity.
Physical Appearance
Sterile Filtered White lyophilized powder
Formulation
Lyophilized from a 0.2µm filtered concentrated (1mg/ml) solution in phosphate-buffered saline (PBS) at pH 7.0.
Solubility
To reconstitute the lyophilized sCD40L, it is recommended to dissolve it in sterile 18 MΩ-cm H2O to a concentration of at least 100 µg/ml. This solution can then be further diluted in other aqueous solutions as needed.
Stability
Lyophilized sCD40L is stable at room temperature for up to 3 weeks. However, it is recommended to store it desiccated at a temperature below -18°C. After reconstitution, CD154 should be stored at 4°C for 2-7 days. For long-term storage, it is advisable to add a carrier protein such as 0.1% HSA or BSA and store it below -18°C. Avoid repeated freeze-thaw cycles.
Purity
The purity of sCD40L is greater than 97.0% as determined by: (a) Reverse-phase high-performance liquid chromatography (RP-HPLC) analysis and (b) Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis.
Biological Activity
The ED50, which represents the concentration of sCD40L required to achieve 50% of the maximum biological response, was determined to be 5-10 ng/ml. This value was obtained by measuring the dose-dependent stimulation of IL-12 induction and IL-8 production by human peripheral blood mononuclear cells (PBMCs).
Protein Content
Protein concentration was determined using two independent methods: 1) UV spectroscopy at 280 nm, using an extinction coefficient of 1.1 for a 0.1% (1 mg/ml) solution. This value was calculated using the PC GENE computer analysis program of protein sequences (IntelliGenetics). 2) Analysis by RP-HPLC, using a standard solution of CD40 as a reference standard.
Synonyms
CD40-L, Tumor necrosis factor ligand superfamily member 5, TNF-related activation protein, TRAP, T cell antigen Gp39, CD154 antigen, sCD40, IGM, IMD3, HIGM1, T-BAM, TNFSF5, hCD40L.
Source
Escherichia Coli.
Product Science Overview
Introduction
Soluble CD-40 Ligand, also known as CD40L, CD154, TRAP (Tumor Necrosis Factor-Related Activation Protein), or TNFSF5, is a member of the Tumor Necrosis Factor (TNF) superfamily. This protein plays a crucial role in the immune system, particularly in the interactions between T cells and antigen-presenting cells (APCs) such as B cells, dendritic cells, and macrophages .
Structure and Expression
CD40 Ligand is a membrane glycoprotein and differentiation antigen expressed on the surface of activated CD4+ T lymphocytes. It can exist in two forms: membrane-bound and soluble. The soluble form is generated through proteolytic cleavage and comprises approximately two-thirds of the extracellular domain .
Biological Functions
The interaction between CD40 Ligand and its receptor, CD40, is essential for various immune responses:
- B Cell Activation and Proliferation: CD40 Ligand stimulates B cells, leading to their activation, proliferation, and differentiation. This interaction is critical for isotype class-switching and the production of different immunoglobulin isotypes .
- Dendritic Cell Maturation: CD40 Ligand promotes the maturation of dendritic cells, enhancing their ability to present antigens and activate T cells .
- Cytokine Production: The interaction induces the production of cytokines such as Interleukin-8 (IL-8), Interferon-gamma (IFN-γ), Tumor Necrosis Factor-alpha (TNF-α), and Interleukin-2 (IL-2) in peripheral blood monocytes and T cells .
- Tumoricidal Activity: CD40 Ligand has been shown to induce tumoricidal activity in peripheral blood monocytes, contributing to the immune system’s ability to target and destroy cancer cells .
Applications in Research
Recombinant human soluble CD40 Ligand is widely used in research for various applications:
- Cell Culture and Differentiation Studies: It is used to stimulate B cell activation and proliferation, as well as dendritic cell maturation .
- Functional Assays: CD40 Ligand is employed in assays to study cytokine production and immune cell interactions .
- Immunotherapy Research: Due to its role in immune activation, CD40 Ligand is investigated for potential therapeutic applications in cancer and autoimmune diseases .
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