Recombinant Biotinylated Human CD40 Ligand / TNFSF5 Protein, Avitag™,His,Flag Tag (active trimer) (MALS verified), 25µg

Background
CD40 ligand is also known as CD40L, CD154, TNFSF5 and T-cell antigen Gp39, is a single-pass type I I membrane protein which belongs to the TNF superfamily of molecules. CD40 ligand is expressed predominantly on activated CD4+ T lymphocytes, and also found in other types of cells, including platelets, mast cells, macrophages, basophils, NK cells, B lymphocytes, as well as non-haematopoietic cells (smooth muscle cells, endothelial cells, and epithelial cells). Although all monomeric, dimeric and trimeric forms of soluble CD40 ligand can bind to CD40, the trimeric form of soluble CD40 ligand has the most potent biological activity through oligomerization of cell surface CD40, a common feature of TNF receptor family members. CD40 ligand binds to CD40 on antigen-presenting cells (APC), which leads to many effects depending on the target cell type. In general, CD40 ligand plays the role of a costimulatory molecule and induces activation in APC in association with T cell receptor stimulation by MHC molecules on the APC. In total CD40 ligand has three binding partners: CD40, α5β1 integrin and αIIbβ3. CD40 ligand regulates B cell function by engaging CD40 on the B cell surface. A defect in this gene results in an inability to undergo immunoglobulin class switch and is associated with hyper IgM syndrome.

Source
Recombinant Biotinylated Human CD40 Ligand, Avitag,His,Flag Tag (CDL-H82Db) is expressed from human 293 cells (HEK293). It contains AA Gly 116 - Leu 261 (Accession # P29965-1).
Predicted N-terminus: Gly

Molecular Characterization
This protein carries a Avi tag (Avitag™) at the N-terminus, followed by flag tag. The polyhistidine tag is inserted in-between the Avitag™ and Flag tags.
The protein has a calculated MW of 51.6 kDa. The protein migrates as 50-55 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.

Biotinylation
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.

Biotin:Protein Ratio
Passed as determined by the HABA assay / binding ELISA.

Endotoxin
Less than 1.0 EU per μg by the LAL method.

Purity
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.

Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.

Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
-20°C to -70°C for 12 months in lyophilized state;
-70^°C for 3 months under sterile conditions after reconstitution.

Bioactivity
Please refer to product data sheet.

Clinical and Translational Updates

(1) "Clinical and Immunological Features, Genetic Variants, and Outcomes of Patients with CD40 Deficiency"
Banday, Nisar, Patra et al
J Clin Immunol (2023) 44 (1), 17
(2) "The CD40/CD40 ligand dyad and its downstream effector molecule ISG54 in relating acute neuroinflammation with persistent, progressive demyelination"
Hazra, Das Sarma
IUBMB Life (2023)
(3) "Novel peptide inhibitors targeting CD40 and CD40L interaction: A potential for atherosclerosis therapy"
Solanki, Kumar, Khan et al
Curr Res Struct Biol (2023) 6, 100110
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