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Ac levd cho

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Ac-LEVD-CHO is a synthetic compound that functions as a caspase-3 inhibitor. It is a tetrapeptide aldehyde that can be used in research applications to study apoptosis and cell death pathways. The core function of Ac-LEVD-CHO is to inhibit the enzymatic activity of caspase-3, which is a key executioner caspase involved in the apoptotic process.

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Lab products found in correlation

Caco 2 tc7 cells, by American Type Culture Collection (2 mentions) Annexin 5 fluorescein isothiocyanate fitc apoptosis detection kit, by BD (1 mentions) Caspase colorimetric assay kit, by Abcam (1 mentions) Z levd fmk, by Enzo Life Sciences (1 mentions) Z atad fmk, by Abcam (1 mentions) Cytochalasin b, by Merck Group (1 mentions) Giemsa stain, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Hoechst 33342, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

3 protocols using ac levd cho

1

Caco-2 TC7 Cell Infection Model

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Caco-2 TC7 cells (ATCC) were seeded at 7.5 x104/ml and upon reaching confluence (7 days) the medium was changed every day for 7 the following 7 days. Before infection the cells were starved for 3 h in serum free DMEM. Monolayers were infected with primed EPEC(20 (link)) at an MOI of 1:10 for 3 h. The cells were then washed twice in PBS and the medium was replaced with serum free DMEM-high glucose plus penicillin and streptomycin at 100 U/ml and 100 μg/ml, respectively. After 1 h cells were washed and either processed for Western Blot (total IL-18) or incubated for a further 17 h (secreted caspase-4 and IL-18) with or without Ac-LEVD-CHO (Enzo Lifesciences). Supernatants were collected, cleared by centrifugation at 13000 rpm at 4 °C for 10 min and precipitated for Western blotting with the addition of 10 % (v/v) trichloroacetic acid for 17 h at 4 ºC. The concentration of IL-18 in cell supernatant (MBL) was determined by ELISA according to the manufacturer’s protocol.
Pallett M.A., Crepin V.F., Serafini N., Habibzay M., Kotik O., Sanchez-Garrido J., Di Santo J.P., Shenoy A.R., Berger C.N, & Frankel G. (2016). Bacterial Virulence Factor Inhibits Caspase-4/11 Activation in Intestinal Epithelial Cells. Mucosal immunology, 10(3), 602-612.
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2

Saikosaponin-Induced Apoptosis Pathway

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Saikosaponin (SSa; Nacalai Tesque Inc., Kyoto, Japan) was dissolved in DMSO (Sigma Chemical Co., St. Louis, MO) to form a 10-mM stock solution and stored at –20°C until use. Propidium iodide (PI), RNase A, Hoechst 33342, cytochalasin-B, Giemsa stain, and 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma. Caspase colorimetric assay kits were purchased from BioVision (Palo Alto, CA). Caspase-4 inhibitors, z-LEVD-fmk and Ac-LEVD-CHO, were obtained from Enzo Life Sciences (Plymouth Meeting, PA). The caspase-12 inhibitor, z-ATAD-fmk, was obtained from BioVision. z-DEVD-fmk (caspase-3 inhibitor), z-VDVAD-fmk (caspase-2 inhibitor), and z-IETD-fmk (caspase-8 inhibitor) were obtained from Calbiochem (La Jolla, CA). The annexin V-fluorescein isothiocyanate (FITC) apoptosis detection kit was purchased from BD Biosciences (Franklin Lakes, NJ, USA).
Kang S.J., Lee Y.J., Kang S.G., Cho S., Yoon W., Lim J.H., Min S.H., Lee T.H, & Kim B.M. (2017). Caspase-4 is essential for saikosaponin a-induced apoptosis acting upstream of caspase-2 and γ-H2AX in colon cancer cells. Oncotarget, 8(59), 100433-100448.
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3

Caco-2 TC7 Cell Infection Model

Check if the same lab product or an alternative is used in the 5 most similar protocols
Caco-2 TC7 cells (ATCC) were seeded at 7.5 x104/ml and upon reaching confluence (7 days) the medium was changed every day for 7 the following 7 days. Before infection the cells were starved for 3 h in serum free DMEM. Monolayers were infected with primed EPEC(20 (link)) at an MOI of 1:10 for 3 h. The cells were then washed twice in PBS and the medium was replaced with serum free DMEM-high glucose plus penicillin and streptomycin at 100 U/ml and 100 μg/ml, respectively. After 1 h cells were washed and either processed for Western Blot (total IL-18) or incubated for a further 17 h (secreted caspase-4 and IL-18) with or without Ac-LEVD-CHO (Enzo Lifesciences). Supernatants were collected, cleared by centrifugation at 13000 rpm at 4 °C for 10 min and precipitated for Western blotting with the addition of 10 % (v/v) trichloroacetic acid for 17 h at 4 ºC. The concentration of IL-18 in cell supernatant (MBL) was determined by ELISA according to the manufacturer’s protocol.
Pallett M.A., Crepin V.F., Serafini N., Habibzay M., Kotik O., Sanchez-Garrido J., Di Santo J.P., Shenoy A.R., Berger C.N, & Frankel G. (2016). Bacterial Virulence Factor Inhibits Caspase-4/11 Activation in Intestinal Epithelial Cells. Mucosal immunology, 10(3), 602-612.
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