Use of sFRP2 and PCPE1 as joint target points in preparation of medicament for fibrotic diseases
A technology for fibrotic diseases and drugs, used in drug combinations, respiratory diseases, biological tests, etc., can solve the problem of unclear interaction of procollagen, and achieve the effect of inhibiting collagen deposition
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[0022] 1. Experimental method
[0023] ① Shear analysis of type Ⅰ procollagen in vitro
[0024] (1) Extraction of radioactive 3H-type procollagen refers to published literature [Scott IC, Blitz IL, Pappano WN, et al. Mammalian BMP-1 / Tolloid-related metalloproteinases, including novel family member mammalian Tolloid-like 2, have differential enzymatic activities and distributions of expression relevant to patterning and skeleton [J]. Dev Biol, 1999, 213(2): 283-300.]. Mix the purified protein 3H-type I procollagen and BMP1, then add 20ng PCPE1, 20ng sFRP2 or 10ng PCPE1 / 10ng sFRP2 respectively, incubate in 100μl buffer A at 37°C for 6h;
[0025] (2) Add SDS sample lysate, heat in a 95°C metal bath for 5 minutes to denature the protein, and then place it on ice to cool for 5 minutes;
[0026] (3) The protein sample was separated by 7.5% SDS-PAGE gel, and then transferred to the membrane;
[0027] (4) Add EN3HANCE (PerkinElmer) and use autoradiography to detect uncut type I pro...
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