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rad140实验引用的文献资料

发布者:RAD140-深圳市华莱生物科技有限公司  发布时间:2017-11-01  阅读:115次
rad140实验引用的文献资料
 Cell assay (Western blots) [2] Cultures are incubated for 5 minutes with 2 μM calcein acetoxymethyl ester and then examined using an
inverted fluorescent IX70 Olympus microscope. The number of healthy, positively stained cells is counted in 4 separate fields (in a
predetermined, regular pattern) per well, 3 wells per condition in each experiment (n ≥ 3 independent culture preparations). Counts of
viable neurons in vehicle–treated controls ranged from 250–300 per well.Lysates are collected from treated cultures using a reducing
sample buffer (62.5 mM Tris–HCl, 1% sodium dodecyl sulfate, 2.5% glycerol, 0.5% 2–β–mercaptoethanol), boiled for 5 minutes, and
centrifuged at 13 000 × g for 10 minutes. The supernatants were analyzed by immunoblotting using a standard protocol with 1 μg/mL
anti–phosphoERK1/2 primary antibody and corresponding horseradish peroxidase–conjugated secondary antibody (1:5000) and
detected using enhanced luminescence. Animal administration [2] GDX rats are administered 1 mg/kg RAD140 suspended in 0.5%
methyl cellulose (1 mg/mL) by daily oral gavage for 2 weeks. This dose was chosen based on previous reports for RAD140 efficacy.
Vehicle–treated animals are gavaged with a similar weight/volume of 0.5% methyl cellulose. On day 13 of the 2–week hormone
treatment period, kainate (10 mg/kg; Enzo Life Sciences) or sterile water control is injected ip. Kainate is dissolved immediately prior to
use in sterile water and lightly heated to fully solubilize. On day 14, SARM–treated rats are administered 1 mg/mL of RAD140
suspended in safflower oil by sc injection because oral gavage is difficult following kainate lesion.
 

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