Cell Signaling Technology

Product Pathways - DNA Damage

Cycloheximide #2112

apoptosis   bortezomib   CHX   cleaved parp   protein synthesis inhibitor   TNF   Tumor Necrosis Factor   ubiquitin  

Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Molecular Characterization

C15H23NO4

Purity

>90%

Structure

Structure

Chemical structure of cycloheximide.放线菌酮的化学结构。

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) , or treated with Cycloheximide (50 μg/ml, 24 hr; +), Bortezomib #2204 (10 nM, 24 hr; +), or both, using Ubiquitin Antibody #3933 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Western blot分析Jurkat细胞提取物,未处理(-)或用放线菌酮(50 μg/ml, 24小时; +),Bortezomib #2204 (10 nM, 24小时; +)处理,或同时用两者处理,使用的抗体是:Ubiquitin Antibody #3933(上图)、β-Actin (D6A8) Rabbit mAb #8457(下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with increasing concentrations of Cycloheximide (24 hr), using PARP Antibody #9542 (upper), Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).Western blot分析Jurkat细胞提取物,未处理(-)或用浓度不断增加的放线菌酮(24小时,+)处理,使用的抗体是:PARP Antibody #9542(上图)、Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625(中间)、β-Actin (D6A8) Rabbit mAb #8457(下图)。

Directions for Use

Cycloheximide is supplied as a lyophilized powder. For a 10 mg/ml stock, carefully weigh out and reconstitute 50 mg in 5 ml DMSO or EtOH. Working concentrations and length of treatments vary depending on the desired effect, but it is typically used at 5-50 µg/ml for 4-24 hours. Soluble in DMSO, EtOH, or MeOH.

提供的放线菌酮为冻干粉。对于10 mg/ml浓度贮存液,在50 mg冻干粉中加入5 ml DMSO或乙醇。工作浓度和处理时间根据所需的预期效果来确定,一般是5-50 µg/ml,处理4-24小时。能溶于DMSO、乙醇和甲醇。

Background

Cycloheximide is a protein synthesis inhibitor in eukaryotes. Although its precise mechanism of action has yet to be fully elucidated, it has been shown to inhibit translation elongation through binding to the E-site of the 60S ribosomal unit and interfering with deacetylated tRNA (1-3). Although not all cell types are equally sensitive to the apoptosis-inducing effects of cycloheximide, it has been shown to induce cell death in T cells through a FADD-dependent mechanism (4). In addition, cycloheximide and Tumor Necrosis Factor possess a synergistic cytotoxicity (5,6), and consequently they are routinely used together to induce cell death. Investigators have demonstrated that cycloheximide blocks bortezomib-stimulated protein ubiquitination (7).

放线菌酮是真核生物的一种蛋白合成抑制剂,其精确的作用机制还需要进一步阐明,研究显示它能通过与60S核糖体的E位点结合和干扰去乙酰基的tRNA来抑制翻译延伸(1-3)。不是所有的细胞对放线菌酮的凋亡诱导效应都有相同的敏感性,研究显示它能通过FADD依赖机制诱导T细胞中的细胞死亡(4)。另外,放线菌酮和肿瘤坏死因子具有协同的细胞毒性(5,6),因此它们常一起使用来诱导细胞死亡。研究者已经证明放线菌酮能阻断硼替佐米刺激的蛋白的遍在蛋白化作用(7)。

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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