Resveratrol, dimethylsulfoxide (DMSO), N-acetyl-L-cysteine (NAC), and methylthiazolyldiphenyl-tetrazolium bromide (MTT) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Terminal deoxynucleotidyl transferase- (TdT-) mediated dUTP nick end labeling (TUNEL) kit was purchased from Roche Inc. (Germany). MitoSOX™ Red mitochondrial superoxide indicator was purchased from Invitrogen (Molecular Probes, Invitrogen, OR, USA). 2′-7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) was purchased from Beyotime Institute of Biotechnology (Jiangsu, China). The antibodies against SOD2, CAT, SULT1A1, and SULT1C2 were purchased from Proteintech (Chicago, IL, USA), and pro-caspase-3, active-caspase-3, pro-caspase-9, and active-caspase-9 from Abcam (Cambridge, UK).
Active caspase 9
Manufactured by Abcam
Sourced in United Kingdom, United States
Active-caspase-9 is an enzyme involved in the process of apoptosis, or programmed cell death. It functions as a critical component of the caspase-dependent apoptotic pathway.
Automatically generated - may contain errors
Lab products found in correlation
Active caspase 3, by Abcam (3 mentions)
Bcl2 associated x (bax), by Abcam (2 mentions)
Pro caspase 3, by Abcam (1 mentions)
Pro caspase 9, by Abcam (1 mentions)
Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
N acetyl l cysteine, by Merck Group (1 mentions)
2 7 dichlorodihydrofluorescein diacetate dcfh da, by Beyotime (1 mentions)
P70s6k, by Merck Group (1 mentions)
Cytochrome c, by Abcam (1 mentions)
Cleaved parp, by Abcam (1 mentions)
Lamp1, by Abcam (1 mentions)
Rapamycin rapa, by Merck Group (1 mentions)
Sqstm1, by Abcam (1 mentions)
P mtor s 2448, by Abcam (1 mentions)
3 methyladenine 3 ma, by Merck Group (1 mentions)
β actin, by Abcam (1 mentions)
Bcl 2, by Abcam (1 mentions)
Beclin 1, by Abcam (1 mentions)
4 protocols using active caspase 9
1
Antioxidant and Apoptosis Modulators
2
Autophagy Regulation by Akt/mTOR Pathway
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P-PDK1(Ser241), Akt, p-Akt(Ser473), 4EBP1 and p-4EBP1(Ser65) and α-tubulin were purchased fromCell Signaling Technology (Danvers, MA, USA). P70S6K and p-P70S6K(Thr470) were acquired from Millipore (Boston, MA, USA). Antibodies against LC3, SQSTM1, Atg5, Beclin1, LAMP1, Bcl-2, Bax, cytochrome C, active caspase-9, active caspase-3, cleaved PARP, mTOR, p-mTOR(S2448) and β-actin were obtained from Abcam (Cambridge, UK). YZT was dissolved in DMSO and diluted with culture medium. CQ, 3-methyladenine (3-MA) and rapamycin (RAPA) were bought from Sigma-Aldrich (Saint Louis, MO, USA).
3
Protein Expression and Quantification
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Cells or tissues were collected and lysed with RIPA buffer (Beyotime), and the total protein was detected using a BCA protein assay kit (Beyotime). The equivalent volume of protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE) and transferred onto the polyvinylidene fluoride membrane. After blocking, the membrane was incubated overnight at 4 °C with primary antibodies, including CyclinD1 (1:1000, Cell Signaling Technology, Danvers, MA, USA), CDK4 (1:1000, Abcam, Cambridge, MA, USA), active caspase 3 (c-caspase 3; 1:1000, Abcam), active caspase 9 (c-caspase 9; 1:1000, Abcam), β-catenin (1:1000, Cell Signaling Technology), and Histone H3 (1:1000, Cell Signaling Technology), DVL2 (1:1000, Abcam), GSK3β (1:1000, Cell Signaling Technology), TCF 4 (1:1000, Abcam), C-MYC (1:1000, Cell Signaling Technology). Anti-β-actin antibody (1:2000, Abcam) acted as an internal control. The membrane was then hybridized with an HRP-conjugated secondary antibody (1:5000, Cell signaling Technology) and the signals were measured using chemiluminescence analysis.
4
Western Blot Analysis of Apoptosis Markers
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Cell lysates collected by radio-immunoprecipitation assay lysis buffer (50 mM Tris pH 7.4, 150 mM NaCl, 2 mM ethylenediaminetetraacetic acid, 1% NP-40, 0.1% sodium lauryl sulfate) were quantified by BCA protein detection method (Pierce, IL, USA). After separation by sulfate polyacrylamide gel electro-phoresis, 30μg of protein was transferred onto polyvinylidene fluoride membrane (Merck Millipore) and incubated with the primary and secondary antibodies. The primary antibodies included Bax (ab32503, Abcam, MA, USA), Bcl-2 (MA5-11950, Invitrogen), Cytochrome C (556433, BD Biosciences, NI, USA), active-caspase-3 (9661, Cell Signaling Technology, MA, USA), active-caspase-9 (ab2324, Abcam), JAK2 (3230, Cell Signaling Technology), p-STAT3 (9145, Cell Signaling Technology), STAT3 (9139, Cell Signaling Technology), GAPDH (60004-1-Ig, Proteintech, IL, USA). The protein membrane was developed by enhanced chemiluminescence (Merck Millipore, Darmstadt, Germany) and photographed by the LAS-4000 imager (GE Healthcare Biosciences, Pittsburgh, PA, USA).
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