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2112
Cycloheximide

Cycloheximide #2112

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).

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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).

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Structure

Chemical structure of cycloheximide.

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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.

Storage:

Store lyophilized or in solution at -20ºC, desiccated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 3 months to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Molecular Weight:

281.3 g/mol

Purity:

>90%

Molecular Formula:

C15H23NO4

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).

  1. Schneider-Poetsch, T. et al. (2010) Nat Chem Biol 6, 209-217.
  2. Klinge, S. et al. (2011) Science 334, 941-8.
  3. Pestova, T.V. and Hellen, C.U. (2003) Genes Dev 17, 181-6.
  4. Mimnaugh, E.G. et al. (2004) Mol Cancer Ther 3, 551-66.
  5. Tang, D. et al. (1999) J Biol Chem 274, 7245-52.
  6. Nolop, K.B. and Ryan, U.S. (1990) Am J Physiol 259, L123-9.
  7. Reid, T.R. et al. (1989) J Biol Chem 264, 4583-9.
For Research Use Only. Not For Use In Diagnostic Procedures.

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

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2112S 1 g $99.00.0
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