Chx-chase assay
WebJan 3, 2024 · When protein biosynthesis is inhibited by cycloheximide (CHX), which prevents translational elongation, it is possible to estimate the protein turnover rate under steady state conditions. Cycloheximide chase assay determined NPNT was a short-lived protein when de novo protein synthesis was blocked ( Figure S1b ), hinting at secretion … WebAug 1, 2015 · CHX chase assay. The half-life of DISC1 protein was evaluated by CHX chase assay using published methods with some modifications (Patrick et al., 1998). Briefly, PC12 cells were seeded into 6-well plates at a density of 3 × 10 5 cells/well in DMEM media containing 10%FBS and 5%HS, and incubated overnight at 37 °C in a CO 2 incubator.
Chx-chase assay
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WebApr 1, 2024 · CHX-chase assay Transfected HEK 293T cells were cultured in presence of 50 μg/ml CHX (Sigma), in some instances in combination with 50 μM MG132 (Calbiochem) for the indicated time periods. WebMay 5, 2024 · Next, a cycloheximide (CHX) chase assay was used to investigate the effect of TRIM25 on RIP3 stability. ... Specific primers used for RT-PCR assays were 5′-ACTACAATACCGCCCACAAC-3′, 5 ...
WebJul 23, 2024 · Cycloheximide-chase assays HEK293 cells were transfected with the indicated expression plasmids for 48 h, and then the cells were treated with 50 µg/mL cycloheximide (CHX). Chronological... WebJan 2, 2024 · CHX chase assay and protein immunoprecipitation (IP) For CHX chase assay, indicated HNSCC cells were treated with 100 μg/ml of CHX for 9 hours maximally. Western blotting was then performed to determine the half-life of the PKM2 protein. For IP assay, the lysed cells were incubated with anti-HA antibody, followed by addition of …
WebSep 7, 2024 · Cycloheximide (CHX) Assay. g at 4°C, then protein samples from cleared whole cell lysates were denatured in 1× Laemmli sample buffer at 95°C for 5 min. Protein samples were resolved on SDS-PAGE gels, transferred to PVDF membranes, then immunoblotting was completed. To determine the protein half-life of wild-type and … WebCycloheximide (CHX; mol. wt. 281.35) is a beige colored, crystal powdered, water-soluble, semisynthetic compound, ... An alternative is the cycloheximide chase assay, where global protein synthesis is blocked using the translation elongation inhibitor cycloheximide. With this method, the clearance of the protein of interest can be ascertained ...
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WebOct 7, 2024 · Cycloheximide (CHX) chase assay was performed to measure RRBP1 degradation in different USP35 expression circumstances. As shown in Fig. 2A , overexpression of wild-type USP35 (WT), but not the catalytic dead mutant (C450A), decreased degradation of RRBP1 in H1299 cells. flixbus vs blablacar busWebApr 1, 2024 · CHX-chase assay. Transfected HEK 293T cells were cultured in presence of 50 μg/ml CHX (Sigma), in some instances in combination with 50 μM MG132 (Calbiochem) for the indicated time periods. For immunoblot read-out, cells were harvested and lysed in NP-40 buffer, followed by immunoblotting. Read-out was by the Odyssey Imaging … great grandchildren in portugueseWebApr 10, 2024 · A cycloheximide (CHX) pulse-chase experiment was then performed to confirm that OTUD6B stabilizes the ... Cycloheximide Pulse-Chase Assay. For the protein half-life assay, the indicated cells were treated with CHX (50 µg mL −1) for the indicated time points, and subjected to immunoblotting. great grandchildren loveWebMay 13, 2011 · Development of dual-reporter cell lines and a CHX chase assay to monitor UPS function. A, HeLa cells stably expressing Ub G76V-GFP and ODD-Luc were treated with the indicated amounts of MG132 for 2 h. Signal intensities for GFP and luciferase accumulation were normalized to values obtained with DMSO-treated cells. B, ... flixbus von paris bercy seineWebApr 1, 2024 · Each SNAP-GPCR-CHX-chase assay was performed in triplicate, and data were analyzed with GraphPad Prism software to calculate the protein half-life (t 1/2) with … great grandchildren in italianWebSep 7, 2010 · CHX files are used for auditing drawings and reporting any violations in an HTML report. They are particularly useful in situations where multiple parties have … great-grandchildren meaningWebChx Chase Assay, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more great-grandchildren definition