膜杰作
Star Staining
分子别名（Synonym）
CD40LG,CD154,CD40L,HIGM1,IGM,IMD3,T-BAM,TNFSF5,TRAP,gp39
表达区间及表达系统（Source）
Human CD40 Ligand Protein, Fc Tag, premium grade (CDL-H5269) is expressed from human 293 cells (HEK293). It contains AA Gly 116 - Leu 261 (Accession # P29965-1) trimer Design.
Predicted N-terminus: Pro
It is produced under our rigorous quality control system that incorporates a comprehensive set of tests including sterility and endotoxin tests. Product performance is carefully validated and tested for compatibility for cell culture use or any other applications in the early preclinical stage. When ready to transition into later clinical phases, we also offer a custom GMP protein service that tailors to your needs. We will work with you to customize and develop a GMP-grade product in accordance with your requests that also meets the requirements for raw and ancillary materials use in cell manufacturing of cell-based therapies.
蛋白结构（Molecular Characterization）
This protein carries a human IgG1 Fc tag at the N-terminus.
The protein has a calculated MW of 74.7 kDa. The protein migrates as 120-150 kDa when calibrated against Star Ribbon Pre-stained Protein Marker under non-reducing (NR) condition (SDS-PAGE) due to glycosylation.
内毒素（Endotoxin）
Less than 0.1 EU per μg by the LAL method.
无菌（Sterility）
The sterility testing was performed by membrane filtration method.
支原体（Mycoplasma）
Negative.
纯度（Purity）
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
制剂（Formulation）
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法（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.
存储（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.
电泳（SDS-PAGE）
Human CD40 Ligand Protein, Fc Tag, premium grade on SDS-PAGE under non-reducing (NR) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).
SEC-MALS
The purity of Human CD40 Ligand Protein, Fc Tag, premium grade (Cat. No. CDL-H5269) is more than 90% and the molecular weight of this protein is around 160-180 kDa verified by SEC-MALS.
Report
活性（Bioactivity）-Bioactivity CELL BASE
Human CD40 Ligand Protein, Fc Tag, premium grade (Cat. No. CDL-H5269) stimulates secretion of IL-6 by human B cells. The ED50 for this effect is 9.68-11.48 ng/mL in the presence of 5 ng/mL of Recombinant Human IL‑4 (Routinely tested).
Protocol
Neutralization assay shows that the cytokine secretion effect of Human CD40 Ligand Protein, Fc Tag, premium grade (Cat. No. CDL-H5269) was inhibited by increasing concentration of anti-CD40L mAb. The concentration of CD40L used is 50 ng/mL. The EC50 is 0.051-0.058 μg/mL (Routinely tested).
Protocol
活性（Bioactivity）-FACS
FACS analysis shows that Human CD40 Ligand Protein, Fc Tag, premium grade (Cat. No. CDL-H5269) can bind to 293T-CD40 cells surface CD40. The concentration of Human CD40 Ligand is 0.3 μg/mL (Routinely tested).
Protocol
活性（Bioactivity）-ELISA
Immobilized Human CD40 Protein, His Tag (Cat. No. CD0-H5228) at 1 μg/mL (100 μL/well) can bind Human CD40 Ligand Protein, Fc Tag, premium grade (Cat. No. CDL-H5269) with a linear range of 0.2-3 ng/mL (QC tested).
Protocol
背景（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.
