分子别名（Synonym）
DIF,TNF-alpha,TNFA,TNFSF2,cachexin,cachectin,TNFα
表达区间及表达系统（Source）
ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (TNA-H4211) is expressed from human 293 cells (HEK293). It contains AA Val 77 - Leu 233 (Accession # P01375-1).
Predicted N-terminus: Val 77
蛋白结构（Molecular Characterization）
This protein carries no "tag".
The protein has a calculated MW of 17.4 kDa. The protein migrates as 17 kDa and 18 kDa under reducing (R) condition (SDS-PAGE).
内毒素（Endotoxin）
Less than 0.1 EU per μg by the LAL method.
纯度（Purity）
>95% as determined by SDS-PAGE.
>95% as determined by SEC-MALS.
制剂（Formulation）
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.
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 6 months under sterile conditions after reconstitution.
电泳（SDS-PAGE）
ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (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%.
SEC-MALS
The purity of ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) was more than 95% in HP-SEC, and the molecular weight of this protein is around 47-60 kDa verified by SEC-MALS.
Report
活性（Bioactivity）-ELISA
Immobilized ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) at 2 μg/mL (100 μL/well) can bind Humira with a linear range of 0.2-2 ng/mL (QC tested).
Protocol
Immobilized ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) at 2 μg/mL (100 μL/well) can bind Human TNFR1, Fc Tag (Cat. No. TN1-H5251) with a linear range of 0.2-3 ng/mL (Routinely tested).
Protocol
活性（Bioactivity）-SPR
Humira (Adalimumab) captured on CM5 chip via anti-human IgG Fc antibodies surface, can bind ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) with an affinity constant of 0.255 nM as determined in a SPR assay (Biacore T200) (Routinely tested).
Protocol
活性（Bioactivity）-Cell based assay
ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) induces cytotoxicity effect on the WEH1-13VAR cells in the presence of the metabolic inhibitor actinomycin D. The EC50 for this effect is 0.007-0.014 ng/mL (Routinely tested).
Protocol
Neutralization assay shows that the cytotoxicity effect of ActiveMax® Human TNF-alpha, Tag Free, low endotoxin (active trimer) (MALS verified) (Cat. No. TNA-H4211) was inhibited by increasing concentration of Adalimumab. The concentration of TNF-alpha used is 1 ng/mL. The IC50 is 29 ng/mL (Routinely tested).
Protocol
文献引用（Citations）
Authors: Hua TNM, et al
Journal: Exp Mol Med 2020
Application: proneural–mesenchymal transition (PMT) assay
Authors: Zhao C,et al.
Journal: J Cell Biochem 2018
Application: ELISA
Authors: Yu L, et al.
Journal: J Cell Physiol 2018
Application: Cell Culture
Diffusion of cytokines in live lymph node tissue using microfluidic integrated optical imaging
Authors: A.E. Ross, et al.
Journal: ANAL CHIM ACTA 2017
Application:
背景（Background）
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.
