Human Tumor Necrosis Factor-alpha (Recombinant)

Recombinant Human Tumor Necrosis Factor-alpha (TNF-alpha)

Source: E.Coli

Background: TNF is secreted by macrophages, monocytes, neutrophils, T-cells, NK-cells following their stimulation by bacterial LPS . Cells expressing CD4 secrete TNF-alpha while CD8 cells secrete little or no TNF-alpha. The synthesis of TNF-alpha is induced by many different stimuli including interferons, IL2 , GM-CSF.

Description: Recombinant Human Tumor Necrosis Factor-alpha produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 158 amino acids and having a molecular mass of 17483.77 Dalton. TNF-alpha is purified by standard chromatographic techniques.

Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation: 1mg of Human TNF-alpha contain 20mM PBS, pH-7.2, and 10mM NaCl.

Solubility: It is recommended to reconstitute the lyophilized Tumor Necrosis Factor-alpha in sterile 18MΩ-cm H₂O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized TNF-alpha although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution TNF-alpha can be stored at 4°C for 2-7 days and below -18 C for longer. It is recommended to add a carrier protein (0.1% HSA or BSA) for long storage. Please prevent freeze-thaw cycles.

Purity: Greater than 98.0% as determined by: (a) Analysis by RP-HPLC. (b) Anion-exchange FPLC. (c) Analysis by SDS-PAGE.

Amino acid sequence: MVRSSSRTPS DKPVAHVVAN PQAEGQLQWL NRRANALLAN GVELRDNQLV VPSEGLYLIY SQVLFKGQGC PSTHVLLTHT ISRIAVSYQT KVNLLSAIKS PCQRETPEGA E AKPWYEPIY LGGVFQLEKG DRLSAEINRP DYLDFAESGQ VYFGIIAL.

Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity: The Specific Activity is >5.0×10⁷ IU/mg as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D.

Protein content: Protein quantitation was carried out by two independent methods:

1. UV spectroscopy at 280 nm using the absorbency value of 1.24 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). 2. Analysis by RP-HPLC, using a calibrated solution of TNF-alpha as a Reference Standard.

Usage: BioWORLD's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.