The rates of dead cells were determined by dye‐exclusion assay using Ho and PI as described previously.31 , 32 Then, 1 × 105 cells/sample were plated on 6‐well plates. After 24 hours, the cells were treated with vehicle (DMSO) or alkylating agents without or with 2 hours pretreatment by pan‐caspase inhibitor Z‐Val‐Ala‐Asp(OMe)‐CH2F (z‐VAD‐FMK; Peptide Institute). The cells were then further cultured for 96 hours and co–stained with Ho and PI. The rate of PI positive cells (dead cells) to Ho positive cells (total cells) was calculated as the cell death rate. Fluorescence images were obtained using a BZ‐X700 microscope (Keyence).
Z val ala asp ome ch2f z vad fmk
Manufactured by Peptide Institute
Sourced in Japan
Z-Val-Ala-Asp(OMe)-CH2F (z-VAD-FMK) is a laboratory reagent used for research purposes. It is a cell-permeable fluoromethyl ketone peptide that inhibits caspase enzymes, which are involved in apoptosis or programmed cell death. The product is used to study cellular processes and mechanisms related to apoptosis in various research applications.
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2 protocols using z val ala asp ome ch2f z vad fmk
1
Quantifying Cell Death Rates
2
Pharmacological Modulation of Glutathione Metabolism
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CDDP, SAS, and DL-propargylglycine (PPG) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Buthionine sulfoximine (BSO) was purchased from Cayman Chemical (Ann Arbor, MI, USA). Z-Val-Ala-Asp(OMe)-CH2F (Z-VAD-FMK) was purchased from Peptide Institute, Inc. (Osaka, Japan). Anti-xCT antibody (ab37185) was purchased from Abcam (Cambridge, UK). Anti-cleaved poly (ADP-ribosyl) polymerase (PARP) antibody (#9541) was purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). Anti-GCLC antibody (GTX16315) was purchased from Gene Tex, Inc. (San Antonio, TX, USA). Anti-cystathionine gamma lyase (CGL) antibody (12217-1-AP) was purchased from Proteintech (San Antonio, TX, USA).
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