Analyze the results by the SDS-PAGE assay and Western blotting (anti-Flag M2 antibodies with 1:5,000 dilution; β-actin antibodies with 1:10,000 dilution). Quantify the Western bands of Flag and β-actin in triplicates using MetaMorph Software or other equivalent softwares.

CHX chase assay visualizes protein degradation kinetics through CHX treatment over different times and western blotting analysis. Briefly, cells are treated with CHX at a suitable concentration, and then harvested at different timepoints. Next, western blot is employed to analyze the expression levels of a given protein.

2014年11月21日 · Cells were treated with CHX for the indicated time and half-lives of c-Myc were determined using western blotting. Quantification of the c-Myc levels is shown.

2023年11月10日 · 用放线菌酮（CHX,50 ng/mL）处理细胞不同时间（0、1、2、4hrs）后，收取细胞进行Western Blotting分析，检测蛋白的变化情况。 我这个western blot蛋白上样量是怎么定的呢？ 每个组取相同体积的蛋白吗？

Among the most frequently used methods to determine protein turnover are pulse chase and cycloheximide (CHX) chase. These two methods examine over time, respectively, the radioisotope-labeled proteins of interest in translation-proficient cells and total preexisting proteins of interest in translation-inhibited cells.

利用 荧光YFP标签 标记目的蛋白，进而利用短暂的闪光脉冲使10-60%的荧光基因失去荧光（即所谓“漂白”）,由于时间短暂因此不会对细胞造成影响。 漂白促使标记蛋白分为了两种亚群：荧光蛋白和非荧光蛋白。 而理论上漂白后细胞不会再合成新的非荧光蛋白，荧光的衰减则只可能是依赖于蛋白质清除。 实验原理是啥呢。 用放线菌酮抑制所有蛋白的合成，隔段时间取样检测目标蛋白的量。 横坐标为时间，蛋白量为纵坐标作图，找出蛋白剩下一半时的时间即为半衰期。 这里要注 …

2023年6月5日 · CHX chase assay visualizes protein degradation kinetics through CHX treatment over different times and western blotting analysis. Briefly, cells are treated with CHX at a suitable concentration, and then harvested at different timepoints. Next, western blot is employed to analyze the expression levels of a given protein. A short half-life

2015年11月4日 · Cycloheximide (Naramycin A) 是一种抗真菌 (antifungal) 抗生素，是真核生物蛋白质合成 (protein synthesis) 的抑制剂，抑制体内蛋白质合成和 RNA 合成的 IC50 值分别为 532.5 nM 和 2880 nM。 Cycloheximide 可抑制铁死亡 (ferroptosis) 并抑制自噬 (autophagy)。 * Please select Quantity before adding items. • Cell. 2025 Feb 6;188 (3):814-831.e21. [Abstract] • Signal Transduct Target Ther. 2024 Sep 26;9 (1):254.

小弟用CHX处理了细胞，分了2h，4h，6h，8h这几个时间梯度，现在有了western图，但是不知道怎么分析。 是用蛋白的灰度值去减内参的灰度还是用蛋白的灰度值除以内参灰度值？

Once protein synthesis is inhibited by CHX, the level of intracellular proteins decreases by degradation through the proteasome or lysosome system. Thus, the CHX chase assay is widely recognized and used to observe intracellular protein degradation and to determine the half-life of a given protein in eukaryotes.