Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, ProClin 300.
Expression System:
E. coli
Formulierung:
Lyophilized powder
Sequenz:
N-terminal His-Tag, The IL35 complex composed of IL27B subunit (Arg21~Lys229 of NP_005746.2) and the mature form of human IL12A (Arg23~Ser219 of NP_000873.2) linked by a polypeptide linker (GGGGS)3
Anwendungsbeschreibung:
IL35 (Interleukin 35) is an IL-12 family cytokine, which is a dimeric protein composed of IL-12alpha and IL-27beta chains. IL35 is thought to mediate the immune inhibitory function of regulatory T cells and has been proven to promotes pancreas cancer growth through enhancement of proliferation and inhibition of apoptosis. Thus, proliferation assay of IL35 was conducted using PANC-1 cells. Briefly, PANC-1 cells were seeded into triplicate wells of 96-well plates at a density of 2000 cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of recombinant human IL35. After incubated for 48h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10 µl of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37C. Proliferation of PANC-1 cells after incubation with IIL35 for 48h observed by inverted microscope. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with human recombinant IL35 for 48h, and IL35 significantly increased cell viability of PANC-1 cells.
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