MABSol® Biotinylated Human TNF-alpha (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 19.2 kDa. The protein migrates as 21 kDa on a SDS-PAGE gel under reducing (R) condition due to glycosylation. More than 95% of Biotinylated Human TNF-alpha (TNA-H82E3) presents as active trimers with a molecular mass of 57 kDa, as determined by SEC-HPLC analysis.
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.
>95% as determined by SEC-HPLC.
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.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 12 months under sterile conditions after reconstitution.
Biotinylated Human TNF-alpha, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95%.
Over 95% of Biotinylated Human TNF-alpha (TNA-H82E3) presents as active trimers with a molecular mass of 57 kDa, as determined by SEC-HPLC analysis.
Immobilized Humira at 5 μg/mL (100 μL/well) can bind Biotinylated Human TNF-alpha, His Tag (Cat. No. TNA-H82E3) with a linear range of 0.4-6 ng/mL (QC tested).
活性（Bioactivity）-Cell based assay
Recombinant Human TNF-alpha (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 0.029-0.052ng/ml (Routinely tested).
Neutralization assay shows that the cytotoxicity effect of TNF-alpha (Cat.No. TNA-H82E3) was inhibited by increasing concentration of Adalimumab. The concentration of TNF-alpha used is 1ng/mL. The IC50 is 7ng/mL (Routinely tested).
Authors: Bourquard T, et al.
Journal: J Immunol 43388
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.
专利使用说明（Limited Use License）
The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the Acrobiosystems’s Avitag™ proteins confers to the purchaser the limited right to use the Avitag™ technology for non-commercial, or research use, or for purposes of evaluating the Avitag™ technology.
Commercial use of the Avitag™ technology to manufacture a commercial product, or use of the Avitag™ technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.