Cytoxicity Bioassay analysis of Bovine IFN-gamma was performed in a viral challenge assay using VS virus on MDBK cells. Various concentrations of Bovine IFN-gamma, ranging from 1.22-2500ng/ml (lyophilized, blue) or 0.12-250ng/ml (solution, red), were added to the cultures. Following incubation, plates were first stained with crystal violet for visual CPE determination, and then the dye was solubilized for absorbance readings. Percentage cytoprotection was determined and compared to cytoprotection achieved with IFN-alpha standard.
|Tested species reactivity||Bovine|
|Host / Isotype||Yeast|
|Storage Conditions||-80° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
For dilution, use a buffer containing carrier protein, e.g. PBS with 1% BSA or a neutral media containing 10% FCS.
Activity: The biological activity of recombinant bovine IFN-gamma was determined in a viral challenge assay using VS virus on MDBK cells. Plates were stained with crystal violet, a visual CPE determination was performed and the dye was then solubilized and absorbance was read. Treatment of MDBK cells with 6 ng/ml of bovine IFN-gamma causes 25% Cytoprotection in the assay.
Expression: The mature form of bovine IFN-gamma was expressed in Pichia pastoris, X-33. GenBank® Accession # AAB64477, amino acids 24 – 166. Refs: Picha, K.S. and P.E. Baker. (1986) Immunol. 57:131. Moscovitch-Lopatin, M. et al. (1991) Eur. J. Immunol. 21:1439.
IFN gamma, also called Type II interferon, macrophage activation factor, and immune interferon is produced primarily by T-lymphocytes and natural killer cells in response to antigens, mitogens, Staphylococcus enterotoxin B, phytohemaglutanin and other cytokines. IFN gamma is a glycoprotein that exists, functionally, as a homodimer of approximately 45 kDa. On SDS-PAGE it appears as a combination of 25, 20 and minor 15.5 kDa bands as a result of differential glycosylation. The biological activity of the IFN gamma homodimer is highly species specific and includes the following: antiviral activity, tumor antiproliferative activity, induction of class I and II MHC, macrophage activation, and enhanced immunoglobulin secretion by B lymphocytes. IFNg is involved in cytokine regulation and also acts synergistically with other cytokines.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.