Cycloheximide pulse chase assay
WebMay 18, 2024 · Cycloheximide is a powerful way to stop further protein production and enable assay of decay rate in vivo ... One way to decrease this effect is to follow the pulse with a chase consisting of a large concentration of unlabelled precursor (the ‘pulse-chase’ approach), which will decrease the chances for re-incorporation of labelled precursor ... WebLuciferase assays were carried out in 96-well white solid bottom plates. d -Luciferin (50 μl of 1 mg/ml in PBS) was added into each well containing 50 μl of medium and incubated at room temperature with shaking at 500 …
Cycloheximide pulse chase assay
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WebCertain lysosomal cathepsin proteins have come into focus as being good candidates for therapeutic targeting, based on them being over-expressed in a variety of cancers and based on their regulation of the apoptotic pathway. Here, we report novel findings that highlight the ability of cathepsin S expression to be up-regulated under Paclitaxel … WebJun 6, 2016 · Pulse-chase As described in the main text, this assay had the following steps, and all liquid handling was performed with electronic multi-channel pipettes or plate washer to improve the...
WebJan 3, 2024 · Cycloheximide pulse chase assay was performed as described previously ( 20 ). Cells were seeded in 6-well plate at 3–5 × 10 5 cells per well. Cells were transfected with EGFP-Nrf2 plasmid and exposed with 50 μg/mL cycloheximide (Sigma). Total protein was extracted and Nrf2 residue was analyzed through Western blot analysis and EGFP … WebTo measure protein half-lives in cultured mammalian cells (HEK293), we used cycloheximide at a concentration of 100 mkg/ml. The control protein which is GFP fused to the C-terminal degron of...
WebBegin the cycloheximide chase as demonstrated for each of the remaining yeast cell suspensions at regular time intervals. At each subsequent time point, vortex the yeast cell suspensions and transfer 950 microliters to labeled microcentrifuge tubes containing 50 microliters of pre-chilled 20x Stop Mix. Vortex and place the collected cells on ice.
WebCycloheximide (CHX) (Sigma Aldrich) is stored as a 500 μg/mL stock solution, solved in dimethylsulfoxide (DMSO) at − 20°C in small aliquots. Proteasome inhibitor MG132 (Enzo Lifesciences) is prepared as a 10 m M stock solution in …
WebCycloheximide is known to inhibit protein synthesis of symbiotic Chlorella of the ciliate P. bursaria preferentially, but it only slightly inhibits host protein synthesis (Ayala and Weis, … hamza yassin new york new york showWebApr 30, 2024 · Cycloheximide Chase This provides a much easier alternative to the pulse-chase and avoids you having to work with radioactivity. Typically, the method … hamza yusuf scotlandWebHere, we identify cyclin-dependent kinase 1 (CDK1) and peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 (PIN1) as two previously uncharacterized regulators of pVHL in multiple types of human cancers harboring wild-type VHLincluding triple … hamza yusuf universityWebhi, I want to ask several questions regarding cycloheximide pulse chase assay. procedure: I want to see if my target protein A extends the half-life of another protein B, so I transfected 293FT cells with B alone (group 1) or A+B (group 2). 48h post transfection, I added cycloheximide at final conc. of 100 ug/ml and lysed cells at 0h, 12h, 24h, 48h … hamz communityWebCycloheximide (CHX) (Sigma Aldrich) is stored as a 500 μg/mL stock solution, solved in dimethylsulfoxide (DMSO) at − 20°C in small aliquots. Proteasome inhibitor MG132 … hamz community websiteWeb这个实验的名字是cycloheximide chase assay（ 环己酰亚胺 ，别称放线菌酮） 实验大致的过程如下 出来的结果是这样子的 咱们再来一张 注意时间点，还有最少有5个时间点。 有 … busabout new zealandWebJan 1, 2015 · Determine the time course of cycloheximide chase beforehand. There can be great variation among proteins. For an unfamiliar protein, it is recommended to start with a 4-hour interval and chase till 24 h. Cycloheximide may cause cytotoxicity to certain cells if cells are exposed to it over 20–24 h. hamza yusuf parents