HEK293T cells stably expressing GABAA receptors were seeded at 2.5 × 105 cells per well in 6-well plates and incubated at 37°C overnight. Cells were then treated with Hispidulin or TP003 for 24 h prior to cycloheximide-chase assay. To stop protein translation, cells were treated with 100 μg/mL cycloheximide, a potent protein synthesis inhibitor (Enzo Life Sciences, catalog #: ALX-380–269). Cells were then chased for the indicated time, harvested, and lysed for SDS-PAGE and Western blot analysis.
Alx 380 269
Manufactured by Enzo Life Sciences
ALX-380–269 is a laboratory instrument used for analytical purposes. It is designed to perform specific functions as part of the research and testing procedures conducted in a laboratory setting. The detailed specifications and intended use of this product are not available for this response.
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Lab products found in correlation
2 protocols using alx 380 269
1
GABA Receptor Stability Assay
2
Transcriptional Response to 20E in Silkworm
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BmN cells were maintained in TC-100 medium (BIOTECH, Germany) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco). Sf9 cell line was cultured in HyClone SFX medium (Thermo Scientific) supplemented with 5% FBS. Cells were pre-incubated for 1 day before further experiments. For ex vivo experiments, newly collected prothoracic gland and fat body tissues from L5D2 larvae were cultured in Grace’s medium (Sigma-Aldrich, 11300-043) at 25 °C. After pre-incubation for 1 h, the medium was replaced with fresh medium. To determine which BmE75 genes are 20E-primary response genes, 2 μM 20E or/and 10 μg/ml CHX (Enzo Life Science, ALX-380-269) were added. After incubation for the indicated times, mRNA and protein were isolated for qPCR analysis and Western blotting.
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