Thermo Fisher Scientific Rat TNF-alpha Recombinant Protein

Applications

Tested Dilution

Publications

Western Blot (WB)

Assay-dependent

-

ELISA (ELISA)

Assay-dependent

-

Immunoprecipitation (IP)

Assay-dependent

-

Control (Ctrl)

Assay-dependent

-

Product Specifications

Species

Rat

Expression System

P. pastoris

Amino acid sequence

Rat TNF-alpha, amino acids 80-235 (Accession # P16599)

Molecular weight

17.2 kDa

Class

Recombinant

Type

Protein

Purity

>95% by SDS-PAGE

Activity

ED50 < 5 pg/mL; determined by the dose-dependent cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D.

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Lyophilized

Purification

purified

Storage buffer

PBS

Contains

no preservative

Storage conditions

4° C

Shipping conditions

Wet ice

Product Specific Information

The mature, soluble form of rat TNF-alpha was expressed in Pichia pastoris, KM71H. GenBank Accession # P16599, amino acids 80 - 235. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: EAEALRSSSQ. Purity is >95% by SDS-PAGE analyses.

Activity: The biological activity of recombinant rat TNFa was measured by its cytolytic effect on mouse WEHI-13VAR cells in the presence of Actinomycin D. The ED50 (or 1 unit of activity) for this effect is typically <5 pg/mL. For in vitro applications a dose-response assay should be performed to determine the optimal concentration for specific experiments.

For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: EAEALSSSQ. The EAEA portion showed incomplete cleavage of the alpha-Factor signal by the Ste 13 protease.

Reconstitution and Dilution: Store lyophilized material at 4ºC. Reconstitute in 0.5 mL of distilled or deionized water. Dilute in a buffer-containing carrier protein, aliquot, and store at or below -70ºC, avoiding multiple freeze-thaw cycles. PBS with 1% BSA or a neutral media containing <10% FCS are suitable buffers.

Target Information

TNF alpha is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophage and bind to its receptors, TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. TNF alpha is involved in the regulation of immune cells, cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS-PAGE under non-reducing conditions. Knockout studies in mice also suggested the neuroprotective function of TNF alpha, and has been observed to causes tumor necrosis when injected into tumor-bearing mice. Other functions of TNF-alpha include its role in the immune response to bacterial, viral, parasitic and certain fungal infections, as well as its role in the necrosis of specific tumors. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon-gamma in its cytotoxicity. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.