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Human TNF-alpha Protein, His Tag

  • Synonym
    DIF,TNF-alpha,TNFA,TNFSF2,cachexin,cachectin,TNFα
  • Source
    Human TNF-alpha, His Tag (TNA-H5228) is expressed from human 293 cells (HEK293). It contains AA Val 77 - Leu 233 (Accession # NP_000585.2).
    Predicted N-terminus: Val 77
  • Molecular Characterization
    Online(Val 77 - Leu 233) NP_000585.2

    This protein carries a polyhistidine tag at the C-terminus.

    The protein has a calculated MW of 18.2 kDa. The protein migrates as 18.2 kDa under reducing (R) condition (SDS-PAGE).

  • Endotoxin
    Less than 1.0 EU per μg by the LAL method.
  • Purity

    >95% as determined by SDS-PAGE.

  • 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.

  • 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.

  • Storage

    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:

    1. 4-8°C for 12 months in lyophilized state;
    2. -70°C for 3 months under sterile conditions after reconstitution.
SDS-PAGE
Human TNF-alpha, His Tag (Cat. No. TNA-H5228) SDS-PAGE gel

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%.

Bioactivity-ELISA
Human TNF-alpha, His TagHuman TNF-alpha, His Tag (Cat. No. TNA-H5228) ELISA bioactivity

Immobilized Human TNF-alpha, His Tag (Cat. No. TNA-H5228) at 0.2 μg/mL (100 μl/well) can bind Human TNFR1, Fc Tag (Cat. No. TN1-H5251) with a linear range of 0.4-6.25 ng/mL (QC tested).

Bioactivity-SPR
Human_FcRn_Heterodimer_Protein_SPR

Humira (Adalimumab) captured on CM5 chip via anti-human IgG Fc antibodies surface, can bind Human TNF-alpha, His Tag (Cat. No. TNA-H5228) with an affinity constant of 0.244 nM as determined in a SPR assay (Biacore T200) (Routinely tested).

Bioactivity-Cell based assay
Human FcRn Heterodimer Protein Cell_Base

Human TNF-alpha, His Tag (Cat. No. TNA-H5228) induces cytotoxicity effect on the WEH1-13VAR cells in the presence of the metabolic inhibitor actinomycin D. The EC50 for this effect is 0.013-0.074 ng/ml (Routinely tested).

Human FcRn Heterodimer Protein Cell_Base

Neutralization assay shows that the cytotoxicity effect of Human TNF-alpha, His Tag (Cat. No. TNA-H5228) was inhibited by increasing concentration of Adalimumab. The concentration of TNF-alpha used is 1 ng/mL. The IC50 is 26.8 ng/mL (Routinely tested).

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