MABSol® Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) (TNA-H82E3) is expressed from human HEK293 cells. It contains AA Val 77 - Leu 233 (Accession # NP_000585.2).
Predicted N-terminus: Val 77
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).
The protein has a calculated MW of 20.0 kDa. The protein migrates as 21 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
The biotin to protein ratio is 0.5-1 as determined by the HABA assay.
Less than 1.0 EU per μg by the LAL method.
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
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 12 months under sterile conditions after reconstitution.
Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
The purity of Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) (Cat. No. TNA-H82E3) was more than 90% in HP-SEC, and the molecular weight of this protein is around 55-70 kDa verified by SEC-MALS.
Immobilized Humira at 5 μg/mL (100 μL/well) can bind Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) (Cat. No. TNA-H82E3) with a linear range of 0.4-6 ng/mL (QC tested).
活性（Bioactivity）-Cell based assay
Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) (Cat.No. TNA-H82E3) induces cytotoxicity effect on the WEH1-13VAR cells in the presence of the metabolic inhibitor actinomycin D. The EC50 for this effect is 0.029-0.052 ng/mL (Routinely tested).
Neutralization assay shows that the cytotoxicity effect of Biotinylated Human TNF-alpha, His,Avitag (active trimer) (MALS verified) (Cat. No. TNA-H82E3) was inhibited by increasing concentration of Adalimumab. The concentration of TNF-alpha used is 1 ng/mL. The IC50 is 7 ng/mL (Routinely tested).
Authors: T Gunde, et al
Journal: US20190153089A1 2019
Authors: Bourquard T, et al.
Journal: J Immunol 2018
Application: Homogeneous time-resolved fluorescence–based competition assay
Authors: Robert Karlsson, et al.
Journal: J Pharmaceut Biomed 2017
Authors: Ji Y, et al.
Journal: J Cell Biochem 2017
Application: Precipitation of TNFR Complexes
Tumor necrosis factor alpha (TNFα) is a cytokine produced primarily by monocytes and macrophages. It is found in synovial cells and macrophages in the tissues.The primary role of TNFα is in the regulation of immune cells. TNFα is able to induce apoptotic cell death, to induce inflammation, and to inhibit tumorigenesis and viral replication. Dysregulation of TNFα production has been implicated in a variety of human diseases, including major depression, Alzheimer's disease and cancer. Recombinant TNFα is used as an immunostimulant under the INN tasonermin. TNFα can be produced ectopically in the setting of malignancy and parallels parathyroid hormone both in causing secondary hypercalcemia and in the cancers with which excessive production is associated.