Introduction
TNF-related apoptosis-inducing ligand (TRAIL), also known as Apo2L, is a protein that acts as a potent inducer of apoptosis (programmed cell death). TRAIL belongs to the TNF superfamily and is characterized as a type II transmembrane protein. In humans, the gene encoding TRAIL is situated on chromosome 3q26. TRAIL demonstrates a specific binding affinity for death receptors DR4 (TRAIL-R1) and DR5 (TRAIL-R2), initiating a caspase-8-dependent apoptotic cascade. This protein exhibits a selective cytotoxicity towards transformed and tumor cells, demonstrating minimal to no effect on normal cells despite its broad expression in various normal tissues.
Description
Recombinant Human TRAIL/APO 2 Ligand, expressed in E. coli, is a single-chain polypeptide. It lacks glycosylation and encompasses 168 amino acids (Met+Arg115-Gly281), resulting in a molecular weight of approximately 21 kDa. The purification of sTRAIL is achieved through proprietary chromatographic techniques.
Physical Appearance
The product appears as a sterile, white powder that has been lyophilized (freeze-dried).
Formulation
The lyophilization process is carried out from a 0.2 μm filtered solution containing 20mM Tris-HCl at pH 8.0 and 150mM NaCl.
Solubility
For reconstitution of the lyophilized TRAIL, it is recommended to use sterile 18 MΩ-cm H2O at a concentration of at least 100 μg/ml. This reconstituted solution can be further diluted as needed using other aqueous solutions.
Stability
Lyophilized APO 2 Ligand, while stable at room temperature for up to 3 weeks, should be stored in a dry environment at temperatures below -18°C. Once reconstituted, TRAIL should be kept at 4°C and used within 2-7 days. For long-term storage, freezing at -18°C is recommended, and the addition of a carrier protein like HSA or BSA (0.1%) is advised. It is important to minimize freeze-thaw cycles.
Purity
The purity of this product exceeds 95.0%, as determined by the following methods: (a) Reverse-phase high-performance liquid chromatography (RP-HPLC) analysis and (b) SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis.
Biological Activity
The biological activity of this product is assessed by its ability to induce cytolysis in Murine L929 cells in the presence of Actinomycin D. The ED50, which represents the concentration required for 50% cell death, is determined to be less than 2 ng/ml. This corresponds to a specific activity greater than 5,000,000 IU/mg.
Synonyms
Tumor necrosis factor ligand superfamily member 10, TNF-related apoptosis-inducing ligand, Protein TRAIL, Apo-2 ligand, Apo-2L, CD253 antigen, TL2, APO2L, TNFSF10.
Amino Acid Sequence
MRERGPQRVA AHITGTRGRS NTLSSPNSKN EKALGRKINS WESSRSGHSF LSNLHLRNGE LVIHEKGFYY IYSQTYFRFQ EEIKENTKND KQMVQYIYKY TSYPDPILLM KSARNSCWSK DAEYGLYSIY QGGIFELKEN DRIFVSVTNE HLIDMDHEAS FFGAFLVG.