TNF a Human, Sf9

Tumor Necrosis Factor-alpha Human Recombinant, Sf9
Cat. No.
BT24973
Source
Sf9, Baculovirus cells.
Synonyms
Tumor Necrosis Factor, TNFA, Tumor Necrosis Factor Ligand Superfamily Member 2, Cachectin, TNF-Alpha, TNFSF2, TNF-A, Tumor Necrosis Factor (TNF Superfamily, Member 2), Tumor Necrosis Factor-Alpha, TNF, Macrophage-Derived, TNF, Monocyte-Derived, TNF Superfamily, APC1 Protein, Member 2, DIF, Tumor necrosis factor.
Appearance
Sterile Filtered clear solution.
Purity
Greater than 90.0% as determined bySDS-PAGE.
Usage
Prospec'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.
Shipped with Ice Packs
In Stock

Description

TNF a produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 163 amino acids (77-233a.a.) and having a molecular mass of 18.1kDa. 
(Molecular size on SDS-PAGE will appear at approximately 18-28kDa).
TNF a is expressed with a 6 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.

Product Specs

Introduction
Tumor necrosis factor (TNF) is a cytokine that plays a key role in systemic inflammation. It belongs to a group of cytokines responsible for initiating the acute phase reaction. Primarily secreted by macrophages, TNF exerts pleiotropic effects, including the induction of apoptotic cell death, stimulation of cellular proliferation and differentiation, modulation of inflammation, involvement in tumorigenesis and viral replication, regulation of lipid metabolism, and participation in coagulation. Notably, TNF plays a crucial role in regulating immune cell function. Dysregulation and excessive production of TNF have been implicated in various human diseases, including autoimmune disorders, insulin resistance, and cancer.
Description
Produced in Sf9 insect cells using a baculovirus expression system, this recombinant TNF-alpha protein is a single, glycosylated polypeptide chain comprising 163 amino acids (residues 77-233). It has a molecular weight of 18.1 kDa, but on SDS-PAGE, it appears as a band at approximately 18-28 kDa due to glycosylation. This TNF-alpha variant is engineered with a 6-amino acid His tag at the C-terminus to facilitate purification, which is achieved through proprietary chromatographic techniques.
Physical Appearance
The product is a clear, sterile-filtered solution.
Formulation
The TNF-alpha protein is supplied as a 1 mg/ml solution in phosphate-buffered saline (PBS) at pH 7.4 with 10% glycerol.
Stability
For short-term storage (up to 4 weeks), the product can be stored at 4°C. For extended storage, it is recommended to store the protein at -20°C. To ensure long-term stability, adding a carrier protein like 0.1% HSA or BSA is advised. Repeated freeze-thaw cycles should be avoided.
Purity
The purity of the TNF-alpha protein is greater than 90%, as determined by SDS-PAGE analysis.
Biological Activity
The biological activity of this TNF-alpha protein was assessed using a cytotoxicity assay with L929 mouse fibrosarcoma cells. The assay measures the ability of TNF-alpha to induce cell death in the presence of actinomycin D, a metabolic inhibitor that enhances TNF-alpha-mediated cytotoxicity. The ED50, which represents the concentration of TNF-alpha required to achieve 50% cell death, is less than or equal to 0.2 ng/ml in this assay.
Synonyms
Tumor Necrosis Factor, TNFA, Tumor Necrosis Factor Ligand Superfamily Member 2, Cachectin, TNF-Alpha, TNFSF2, TNF-A, Tumor Necrosis Factor (TNF Superfamily, Member 2), Tumor Necrosis Factor-Alpha, TNF, Macrophage-Derived, TNF, Monocyte-Derived, TNF Superfamily, APC1 Protein, Member 2, DIF, Tumor necrosis factor.
Source
Sf9, Baculovirus cells.
Amino Acid Sequence
VRSSSRTPSD KPVAHVVANP QAEGQLQWLN RRANALLANG VELRDNQLVV PSEGLYLIYS QVLFKGQGCP STHVLLTHTI SRIAVSYQTK VNLLSAIKSP CQRETPEGAE AKPWYEPIYL GGVFQLEKGD RLSAEINRPD YLDFAESGQV YFGIIALHHH HHH.

Product Science Overview

Introduction

Tumor Necrosis Factor-alpha (TNF-α) is a potent pro-inflammatory cytokine that plays a crucial role in the immune system. It is involved in systemic inflammation and is one of the cytokines that make up the acute phase reaction. TNF-α is produced primarily by macrophages, but it can also be produced by other cell types such as lymphocytes, natural killer cells, and neurons.

Structure and Expression

The human recombinant TNF-α produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 163 amino acids, with a molecular mass of approximately 18.1 kDa . The recombinant protein is expressed with a 6 amino acid His tag at the C-terminus, which facilitates its purification through chromatographic techniques .

Biological Activity

TNF-α is a key regulator of immune cells and has a wide range of biological activities. It can induce fever, apoptotic cell death, sepsis (through IL-1 & IL-6 production), cachexia, inflammation, and inhibit tumorigenesis and viral replication . The recombinant TNF-α produced in Sf9 cells retains these biological activities and is often used in research to study these processes.

Applications in Research

Recombinant TNF-α is widely used in scientific research to study its role in various diseases, including rheumatoid arthritis, psoriatic arthritis, and psoriasis . It is also used to investigate the mechanisms of inflammation and immune response, as well as to develop and test new therapeutic agents targeting TNF-α.

Storage and Stability

The recombinant TNF-α protein is typically stored at 4°C if it will be used within 2-4 weeks. For longer storage, it is recommended to keep the protein frozen at -20°C, with the addition of a carrier protein such as 0.1% HSA or BSA to prevent degradation . It is important to avoid repeated freeze-thaw cycles to maintain the protein’s stability and activity.

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