Rat Tumor Necrosis Factor-alpha (Recombinant)

Recombinant Rat 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 rat TNF-alpha produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 157 amino acids and having a molecular mass of 17339.44 Dalton. Recombinant Rat TNF-alpha is purified by standard chromatographic techniques.

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

Formulation: The protein was lyophilized with no additives.

Solubility: It is recommended to reconstitute the lyophilized Rat TNF-alpha in sterile 18MO-cm H2O not less than 100 g/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized Rat TNF-alpha although stable at room temperature for 3 weeks, should be stored desiccated below -180C. Upon reconstitution Rat TNF-alpha should be stored at 40C between 2-7 days and for future use below -180C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.

Purity: Greater than 97.0% as determined by: (a) Analysis by RP-HPLC. (b) Anion-exchange FPLC. (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Leu-Arg-Ser-Ser.

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

Biological Activity: ProSpec\\\'s Rat TNF-alpha is fully biologically active when compared to standard. The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D is < 0.05ng/ml, corresponding to a Specific Activity of 5x107 IU/mg.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of Rat TNF-alpha.

Protein content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm using the absorbency value of 1.25 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-a 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.