ADI (lot number: 20180522) was supplied by Guizhou Ebay Pharmaceutical Co., Ltd. (Guizhou, China). DOX (purity > 98%) and the internal standard (IS), azithromycin (purity > 98%), were purchased from Dalian Meilun Biotech Co., Ltd. (Liaoning, China). DOXol was obtained from Glpbio (Montclair, CA, USA). Hoechst 33342 was purchased from Beyotime Institute of Biotechnology (Haimen, China). Aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatinine kinase (CK), CK-MB isoenzyme and caspase-3 activity assay kits were provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). A mouse BNP (brain natriuretic peptide) ELISA kit was purchased from Shanghai MLBIO Biotechnology Co., Ltd. (Shanghai, China). BCA assay kit, CBR1 and GAPDH antibody were purchased from Abcam (Cambridge, UK). High performance liquid chromatography (HPLC)-grade acetonitrile, methanol and formic acid were supplied by Merck Company Inc. (Darmstadt, Germany). Distilled water was obtained from Watsons Group Co., Ltd. (Hong Kong, PRC). All other chemicals and reagents used were of chromatographic or analytical grade.
Caspase 3 activity assay kit
Manufactured by Nanjing Jiancheng
Sourced in China
The Caspase-3 activity assay kit is a laboratory tool used to measure the activity of the enzyme Caspase-3. Caspase-3 is a key mediator of apoptosis, a form of programmed cell death. The kit provides the necessary reagents and protocols to quantify the enzymatic activity of Caspase-3 in cell or tissue samples.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Caspase 9 activity assay kit, by Nanjing Jiancheng (2 mentions)
Ck mb isoenzyme, by Nanjing Jiancheng (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Elisa kit, by Mlbio (1 mentions)
Aspartate aminotransferase, by Nanjing Jiancheng (1 mentions)
Lactate dehydrogenase, by Nanjing Jiancheng (1 mentions)
Azithromycin, by Meilun (1 mentions)
Malondialdehyde mda detection kit, by Nanjing Jiancheng (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
In situ apoptosis detection kit, by Roche (1 mentions)
Ca2 atpase assay kit, by Nanjing Jiancheng (1 mentions)
A spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Annexin 5 pi kit, by BD (1 mentions)
Cell mda assay kit, by Nanjing Jiancheng (1 mentions)
Bradford protein assay kit, by Nanjing Jiancheng (1 mentions)
Horse serum, by Solarbio (1 mentions)
Glutathione reductase assay kit, by Nanjing Jiancheng (1 mentions)
Ldh cytotoxicity kit, by Nanjing Jiancheng (1 mentions)
Gentamicin, by Solarbio (1 mentions)
18 protocols using caspase 3 activity assay kit
1
Pharmacokinetic Analysis of Doxorubicin
2
Oxidative Stress and Apoptosis Assays
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ALA, paraformaldehyde, and 2,3,5-triphenyltetrazolium chloride (TTC) were purchased from Sigma-Aldrich (MO, USA). Tissue protein extraction kits, the bicinchoninic acid assay (BCA) kits, the primary antibodies, and the respective secondary antibodies were purchased from Santa Cruz Biotechnology (CA, USA). The malondialdehyde (MDA) detection kits and caspase-3 activity assay kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The ELISA kits for protein carbonyl and 8-hydroxydeoxyguanosine (8-OHdG) were purchased from Cell Biolabs (CA, USA).
3
Caspase 3 Activity Quantification
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The activity of caspase 3 was measured using caspase 3 activity assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) according to the manufacturer’s protocol. The assay is based on spectrophotometric detection of the chromatophore ρ-nitroaniline (ρNA) after cleavage from the labeled substrate Ac-DEVD-ρNA by caspase 3 protease. The level of caspase 3 activity was tested at 405 nm in a microplate reader. Fold increase in caspase 3 activity was determined by comparing the absorbance from an apoptotic sample with an untreated control after subtracting the background value reading from cell lysates and buffers.
4
Measuring Myocardial Apoptosis and Caspase-3
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Fresh heart samples were collected and assayed for myocardial apoptotic cell death using terminal deoxynucleotidyl transferase-meditated dUTP nick-end labeling (TUNEL) staining with an in situ apoptosis detection kit (Roche Diagnostics Ltd., USA). Fresh heart samples were homogenized, and the activity of caspase3 in the heart was examined using a caspase3 activity assay kit from Nanjing Jiancheng Bioengineering Institute.
5
Biochemical Markers in Cellular Processes
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Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were determined using an automatic biochemical analyzer (Chemray 80, Shenzhen, China). Intracellular SERCA activity was measured using a Ca2+-ATPase assay kit (Jiancheng, Nanjing, China) and normalized to protein concentration. The caspase-3 activity assay kit (Jiancheng, Nanjing, China) was used to detect the activity of caspase-3 enzyme in cell and tissue lysates.
6
Apoptosis in Liver Cancer Cells
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The apoptosis in HepG2 cells and primary hepatoma carcinoma cells was detected by flow cytometry (FCM) using the Annexin V: PI kit (BD, Franklin lakes, New Jersey, USA, 556463, 560931). Briefly, the cells were collected and washed twice with cold PBS. The cells were incubated in 100 μL of binding buffer with 5 μL Annexin V and 10 μL PI in the dark at room temperature for 30 min. The samples were analyzed by FCM (Beckman Coulter, Miami, FL). The caspase 3 activity of HepG2 cells was detected using caspase 3 activity assay kit (Jiancheng, China). Briefly, the cells were collected and washed twice with cold PBS. The cells were resuspended and lysed on ice for 30 min. The supernatant was incubated with Ac‐DEVD‐pNA and reaction buffer at 37°C for 4 h. The absorbance at 405 nm was recorded with a spectrophotometer (Thermo Scientific).
7
Antioxidant and Apoptotic Effects of Natural Compounds
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Poly-D-lysine hydrobromide, ferulic acid (99%), vanillin (99%), vanillic acid (97%), 3-(4,5-dimethylthialzal-z-yl)-2,5-diphenylterazolium (MTT), DCFH-DA probes, and Nutrient Mixture F-12 Ham were purchased from the Sigma-Aldrich Shanghai Trading Co. (Shanghai, China). Curcumin (98%) was obtained from the Aladdin Industrial Corporation (Shanghai, China). Fetal bovine serum (FBS) was bought from (Gibco BRL, Grand Island, NY, USA). Horse serum was bought from Solarbio Biological Technology Co., Ltd. (Beijing, China). Hydrogen peroxide (H2O2) was obtained from Shuangshuang Chemical Co., Ltd. (Yantai, China). An annexin V-FITC/PI apoptosis detection kit was obtained from Bibo Biological Technology Co., Ltd. (Nanjing, China). A Bradford protein assay kit, caspase-3 activity assay kit, caspase-9 activity assay kit, and cell MDA assay kit were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All other reagents used were of analytical grade.
8
Caspase-3 Activity Assay Protocol
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A caspase-3 activity assay was performed using a colorimetric Caspase-3 Activity Assay kit (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), according to the manufacturer's protocol. Briefly, 200 µg tissue lysate was combined with 100 µl reaction buffer containing 5 µl caspase-3 substrate DEVD-pNA (4 mM), 1% NP-40, 20 mM Tris-HCl (pH 7.5), 137 mM n-acetyl-cysteine and 10% glycerol. The lysates were incubated at 37°C for 2 h in the dark and the absorbance was measured at a wavelength of 405 nm using a microplate reader, as previously described (23 (link)).
9
Assaying Cellular Cytotoxicity and Apoptosis
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The LDH release, GSH content, caspase-3 activity and caspase-9 activity were determined using the LDH cytotoxicity kit, glutathione reductase assay kit, caspase-3 activity assay kit and caspase-9 activity assay kit (Nanjing JianCheng Bio Institute, Nanjing, China), respectively, according to the manufacturer protocol [18 (link),27 (link),28 (link)].
10
Investigating Cardiomyocyte Stress Responses
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N-acetylcysteine (NAC; antioxidant), PA, 4-phenylbutyrate (4-PBA; ER stress inhibitor), and bovine serum albumin (BSA) were purchased from Sigma–Aldrich (St. Louis, MO, U.S.A.). Fetal bovine serum (FBS) was purchased from Gibco (Grand Island, NY, U.S.A.). Dulbecco’s modified Eagle’s medium (DMEM) was obtained from HyClone (Logan, Utah, U.S.A.). H9c2 cardiomyocytes were purchased from the Chinese Academy of Sciences (Shanghai, China). A caspase 3 Activity Assay Kit was purchased from the NanJing JianCheng Bioengineering Institute (Nanjing, Jiangsu, China). NaOH, phosphate-buffered saline (PBS), penicillin, and gentamicin were obtained from Solarbio (Beijing, China). A CCK8 Assay Kit, ROS Assay Kit, and Annexin V-PE/PI Apoptosis Analysis Kit were purchased from the Beyotime Institute of Biotechnology (Shanghai, China). TRIzol and a PrimeScript® RT Reagent Kit were purchased from Invitrogen (Carlsbad, CA, U.S.A.). PVDF membranes were obtained from Millipore (Bedford, MA).
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