Web8 de jun. de 2024 · Autophagy-mediated long lived proteins degradation assay We adopted a well-described protein degradation assay30,31 for use in our differentiated epididymal pre-adipocyte cells. Five days after differentiation induction, cells were ‘pulsed’ for 24 h with 50 µCi/ml L-[14C(U)]-Valine (NEC291EU050UC, PerkinElmer), then washed Web5 de mai. de 2024 · The Long-lived Protein Degradation Assay: an Efficient Method for Quantitative Determination of the Autophagic Flux of Endogenous Proteins in Adherent …
Interaction of TBC1D9B with Mammalian ATG8 Homologues …
WebKeywords: Long-lived protein degradation, Autophagy, Autophagic flux, Endogenous cargo, Quantitative assay, Pulse-chase, Valine, 14C radioactivity [Background] Pulse-chase labelling approaches have been used to study protein turnover for decades. In the longlived protein degradation (LLPD) assay described here, the proteome of cells in … Web2 de fev. de 2016 · To determine if a functional relationship exists between α-syn accumulation and the lysosomal system in human midbrain dopamine (DA) neurons, we measured the degradation rate of long-lived proteins in neurons that overexpress α-syn by lentiviral transduction. symptoms in broad problems examples
Quantitative Cell-based Protein Degradation Assays to Identify …
Web10 de fev. de 2024 · Long-lived protein degradation assay Cells were incubated for 36 h with 0.135 μCi/ml L-[ 14 C]valine supplemented complete RPMI (with 10% FBS and 1% penicillin and streptomycin) medium, followed by two washes and a 3 h chase for degradation of short-lived proteins in fresh medium containing 10 mM nonradioactive L … WebHowever, traditional methods, such as a radioisotope labeling assay, are technically tedious and have low sensitivity. Here, we report a novel method for quantification of long-lived … Web10 de set. de 2024 · Long-lived protein degradation assay was performed as previously described 48. Briefly, cells were cultured in DMEM in 24-well plates. L-[ 14 C]-valine was added to a final concentration of 0.2 ... thai development pcl. itd ที่อยู่