2019年1月15日 · To understand if the decreased AGR2 by MG132 is caused by protein stability, cycloheximide (CHX), a protein synthesis inhibitor, was included in the study below.

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 …

The whole cell lysate input was probed using anti-FLAG antibody to determine the expression of flag-ELL2. GAPDH was detected as loading control.

通过CO-IP、免疫荧光探究USP22和NLRP3蛋白之间的关系；使用CHX、MG132、CQ处理Flag标记的USP22和HA-NLRP3共转染的293T细胞，探究USP22降解NLRP3蛋白的方式；通过构建EBSS或雷帕霉素诱导的USP22敲除的细胞自噬模型，采用WB、免疫荧光法探究USP22对自噬的影响；通过CO-IP、WB、免疫荧光、ELISA探究USP22影响自噬作用的分子机制；最后通过点突变+CO-IP+泛素化实验对蛋白-蛋白的结合位点进行解析。

GAPDH，全称（glyceraldehyde-3-phosphate dehydrogenase）或者甘油醛-3-磷酸脱氢酶，实际上它是糖酵解反应中的一种酶，在所有组织细胞中都呈现高表达，在同一细胞或者组织中表达量基本恒定，很少受外部诱导物的影响。 少见于缺氧环境或者糖尿病及其他疾病情况下，GAPDH 在特定细胞和组织中有表达. 如何跑一条漂亮的内参. WB做的是否完美，一条整齐的内参条带是必要因素之一。 有多少小伙伴因为内参不整齐反复调整来回跑WB到秃头~，实际上跑内参也是有小 …

2011年1月4日 · Inhibition of new protein synthesis by cycloheximide (CHX) can block the survival pathway and, thus, is widely used in studying cell death. Indeed, treatment of primary MEF cells with 30 ng/ml...

1990年7月16日 · GAPDH mRNA is induced two- to fivefold over resting levels upon IL2 stimulation, due in part to an increased rate of transcription. GAPDH enzymatic activity is induced approx. sevenfold over resting levels. The induction of GAPDH mRNA is inhibited only slightly by CHX under conditions in which cell proliferation is inhibited.

2021年7月27日 · Here, we show that the m 6 A reader YTHDF3 is essential for autophagy induction. m 6 A modification is up-regulated to promote autophagosome formation and lysosomal degradation upon nutrient...

2016年7月4日 · By measuring the turnover of the well-characterized CMA substrate, GAPDH, following protein synthesis inhibition with cycloheximide (CHX) for 24h, we detected that GAPDH protein levels were significantly decreased by 49.6% in Gba1 WT MEF following CHX treatment, whereas GAPDH protein levels in Gba1 HET and Gba1 KO MEF were similar to non ...

2023年6月12日 · Homologous recombination (HR), a form of error‐free DNA double‐strand break (DSB) repair, is important for the maintenance of genomic integrity. Here, we identify a moonlighting protein, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), as a regulator of HR repair, which is mediated through HDAC1‐dependent regulation of RAD51 stability.