Intracellular isoform of the interleukin-1 receptor antagonist
An interleukin and antagonistic technology, applied in the field of biology, can solve the problems of unclear biological significance
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[0067] Cloning and Characterization of icIL-1ra II
[0068] Materials and methods
[0069] Reagent
[0070] The following commercially available reagents are used for cell culture and isolation: pyrogen-free saline and distilled water for clinical use: RPMI 1640 medium; DMEM medium; M199 medium; L-glutamine; Percoll; Ficoll-Hipaque; Aseptically collected fetal bovine serum; Endothelial cell growth supplement (ECGS) prepared from bovine brain; Heparin.
[0071] The endotoxin contained in all reagents is less than 0.125EU / ml, which is checked by limulus amebocyte lysate test.
[0072] cell
[0073] Human circulating PMN and mononuclear cells are separated by isotonic (285mOsm) Perke discontinuous (46% for monocytes and 62% for PMN) gradient centrifugation from the peripheral blood of healthy donors, such as Colotta F., Peri G., The method described in Villa SA., Mantovani A., Rapid killing of actinomycin D treated tumor cells by human mononuclear cells. J. Immunol. 132:936,1...
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