TNF a Human, His
Greater than 97.0% as determined by SDS-PAGE and HPLC analyses.
Description
Tumor Necrosis Factor-α Human Recombinant His produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 164 amino acids fragment and having a molecular mass of 18.3kDa with an N-terminal hexahistidine tag. The TNF-alpha His is purified by standard chromatographic techniques.
Product Specs
Tumor necrosis factor (TNF), a cytokine, plays a crucial role in systemic inflammation and belongs to a family of cytokines that initiate the acute phase reaction. Primarily produced by macrophages, TNF exerts pleiotropic effects on cells, including the induction of apoptosis, stimulation of proliferation and differentiation, modulation of inflammation, involvement in tumorigenesis and viral replication, and regulation of lipid metabolism and coagulation. Notably, TNF plays a central role in regulating immune cell function. Dysregulation and excessive production of TNF are implicated in various human diseases, including autoimmune disorders, insulin resistance, and cancer.
Recombinant human Tumor Necrosis Factor-alpha (TNF-α) with a His tag, expressed in E. coli, is a single, non-glycosylated polypeptide chain. This protein fragment consists of 164 amino acids, has a molecular weight of 18.3 kDa, and includes an N-terminal hexahistidine tag. The purification process of TNF-alpha His involves standard chromatographic techniques.
The product appears as a sterile, filtered, and lyophilized (freeze-dried) powder, white in color.
The product is lyophilized from a 0.2 µm filtered concentrated solution prepared in phosphate-buffered saline (PBS) at a pH of 7.0.
For reconstitution of the lyophilized TNF-α, it is recommended to use sterile 18 MΩ-cm H₂O at a concentration not less than 100 µg/ml. This solution can be further diluted into other aqueous solutions as required.
Lyophilized TNF-α, while stable at room temperature for up to 3 weeks, should be stored desiccated at a temperature below -18°C. After reconstitution, TNF-α should be stored at 4°C for a period of 2-7 days. For long-term storage, it is advisable to add a carrier protein (0.1% HSA or BSA). Freeze-thaw cycles should be avoided.
The purity of the product is determined to be greater than 97.0% based on SDS-PAGE and HPLC analyses.
The ED50, determined by a cytotoxicity assay using murine L929 cells in the presence of actinomycin D, is less than 0.05 ng/ml. This corresponds to a specific activity greater than 2.0 × 10⁷ IU/mg.
MHHHHHHVRS SSRTPSDKPV AHVVANPQAE GQLQWLNRRA NALLANGVEL RDNQLVVPSE GLYLIYSQVL FKGQGCPSTH VLLTHTISRI AVSYQTKVNL LSAIKSPCQR ETPEGAEAKP WYEPIYLGGV FQLEKGDRLS AEINRPDYLD FAESGQVYFG IIAL.
Product Science Overview
The recombinant human TNF-α protein with a His tag is typically expressed in Escherichia coli (E. coli) or Chinese Hamster Ovary (CHO) cells . The His tag, usually consisting of six histidine residues, is added to the N- or C-terminus of the protein to facilitate purification through affinity chromatography . The recombinant protein is often produced as a non-glycosylated polypeptide chain with a molecular mass of approximately 18.3 kDa .
Recombinant TNF-α proteins are characterized by high purity levels, often exceeding 95% as determined by SDS-PAGE . The endotoxin levels are kept very low, typically below 1.000 EU/µg, to ensure the protein’s safety and efficacy in research applications . The bioactivity of the recombinant protein is maintained, making it suitable for various experimental setups .
TNF-α is widely used in research to study its role in inflammation, immune response, and cell signaling pathways. It is particularly valuable in investigating the mechanisms of diseases such as rheumatoid arthritis, Crohn’s disease, and cancer . The His tag allows for easy purification and detection of the protein, making it a versatile tool in molecular biology and biochemistry research .
Recombinant TNF-α proteins are typically lyophilized and can be reconstituted in PBS (phosphate-buffered saline) for use . They should be stored at -25 to -15°C for long-term storage and at 2-8°C for short-term use after reconstitution . It is recommended to aliquot the protein into smaller quantities to avoid repeated freeze-thaw cycles, which can degrade the protein .
