A kind of autophagy inhibitory polypeptide
An autophagy inhibition, cell technology, applied in the field of bioengineering, can solve the problems of poor prognosis of patients, low specificity of chloroquine inhibiting autophagy, limiting the clinical application of chloroquine, etc. Conducive to the effect of large-scale promotion and utilization
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[0022] 1. The interaction test between FIP200 and its truncation mutants and ATG13 protein, the purpose is to identify the minimum structural domain that potentially mediates the mutual requirement of FIP200 and ATG13 protein:
[0023] Main experimental materials:
[0024] 1) cells
[0025] 293T cells were selected in this experiment. The cell culture is convenient, the growth is fast, and the autophagy activity of liposome transfection in vitro is high. In the case of autophagy induction, the interaction between FIP200 and ATG13 protein can be significantly detected by co-immunoprecipitation, and the autophagy The marker LC3ii band is easier to detect.
[0026] 293T, cells, purchased from ATCC company, culture medium DMEM Gibico, containing 10% fetal bovine serum life tech company, cultivated in a cell incubator at 5% CO2, 37 degrees, 95% humidity, 3), antibody, flag antibody sigma, 1:2000, HA-beads sigma, GFP antibody, FIP200 antibody, LC3 antibody, cell signal technology...
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