Dulbecco’s modified Eagle’s medium low glucose (DMEM-LG), fetal bovine serum (FBS), trypsin, dexamethasone (Dex), β-glycerophosphate (β-GP) and vitamin C (vit C) were purchased from Gibco (Grand Island, NY). U0126, antibodies specific for ERK1/2, phospho-ERK1/2, phospho-p38-MAPK monoclonal antibody, IgG/HRP and β-actin polyclonal antibody were purchased from Cell Signaling Technology (Beverly, MA). Gomori staining kit, alizarin red dye solution, rat collagen-I (Col-I) and ELISA detection kit were obtained from the Jiancheng Bioengineering Institute (Nanjing, China). TRIzol was purchased from Invitrogen (Carlsbad, CA). Primer synthesis and RT, PCR kits were purchased from Sangon Biotech (Shanghai, China). The CCK-8 kit and Western blotting reagents were purchased from Beyotime Biotechnology Institute (Haimen, China).
Western blotting reagents
Manufactured by Beyotime
Sourced in China
Western blotting reagents are a set of laboratory tools used to detect and quantify specific proteins in a sample. The core function of these reagents is to facilitate the process of protein separation, transfer, and detection through techniques such as electrophoresis and immunodetection.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Alizarin red dye solution, by Nanjing Jiancheng (1 mentions)
Elisa detection kit, by Nanjing Jiancheng (1 mentions)
β actin polyclonal antibody, by Cell Signaling Technology (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Cck 8 kit, by Beyotime (1 mentions)
Gomori staining kit, by Nanjing Jiancheng (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Primary antibodies against lc3b, by Cell Signaling Technology (1 mentions)
Type 2 collagen, by Cell Signaling Technology (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Immunofluorescence reagents, by Beyotime (1 mentions)
β galactosidase staining kit, by Beyotime (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Ki 67, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
6 protocols using western blotting reagents
1
Osteogenic Differentiation Protocol
2
Chondrocyte Autophagy and Differentiation
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Alcian dye, rat-derived recombinant PTH and type II collagenase were purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). Primary antibodies against LC3b, Beclin-1, P62, ATG5, type II collagen, SOX-9 and β-actin were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). β-galactosidase staining kits, immunofluorescence reagents, western blotting reagents, DAPI, and cell protein extraction kits were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Finally, cell culture reagents, including Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS) and 0.25% trypsin were purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA).
3
Autophagy and Apoptosis Regulation in Cells
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MTT, crystal violet solution, PI, RNAse A and DAPI were purchased from Beijing Solarbio Science & Technology Co., Ltd. Western blotting reagents were obtained from Beyotime Institute of Biotechnology. An Annexin V/PI apoptosis detection kit was purchased from 7 Sea Biotech (http://www.7seapharmtech.com/ ).
Primary antibodies against the following proteins were purchased from Cell Signaling Technology, Inc. (CST): AKT (cat. no. 9272), phosphorylated (p)-AKT (Ser473; cat. no. 4060), mTOR (cat. no. 2983), p-mTOR (Ser2448; cat. no. 5536), p70S6K (cat. no. 9202), p-p70S6 kinase (p70S6K; Ser371; cat. no. 9208), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1; cat. no. 9644), p-4E-BP1 (Thr37/46; cat. no. 2855), Beclin-1 (cat. no. 3495), p-Beclin-1 (Ser93; cat. no. 14717), Cyclin D1 (cat. no. 2978), Cyclin E1 (cat. no. 4129), cleaved caspase-3 (cat. no. 9664), autophagy related 5 (ATG5; cat. no. 9980) and GAPDH (cat. no. 5174). LC3-I/II (cat. no. ABC929) was purchased from Sigma-Aldrich (Merck KGaA) and ki67 (cat. no. ab15580) was obtained from Abcam. Secondary horseradish peroxidase-linked antibodies against rabbit (cat. no. 7074) and mouse (cat. no. 7076) were purchased from CST. Small interfering RNA (si)ATG5 was purchased from Shanghai GenePharma Co., Ltd., and Lipofectamine® 2000 was obtained from Thermo Fisher Scientific, Inc.
Primary antibodies against the following proteins were purchased from Cell Signaling Technology, Inc. (CST): AKT (cat. no. 9272), phosphorylated (p)-AKT (Ser473; cat. no. 4060), mTOR (cat. no. 2983), p-mTOR (Ser2448; cat. no. 5536), p70S6K (cat. no. 9202), p-p70S6 kinase (p70S6K; Ser371; cat. no. 9208), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1; cat. no. 9644), p-4E-BP1 (Thr37/46; cat. no. 2855), Beclin-1 (cat. no. 3495), p-Beclin-1 (Ser93; cat. no. 14717), Cyclin D1 (cat. no. 2978), Cyclin E1 (cat. no. 4129), cleaved caspase-3 (cat. no. 9664), autophagy related 5 (ATG5; cat. no. 9980) and GAPDH (cat. no. 5174). LC3-I/II (cat. no. ABC929) was purchased from Sigma-Aldrich (Merck KGaA) and ki67 (cat. no. ab15580) was obtained from Abcam. Secondary horseradish peroxidase-linked antibodies against rabbit (cat. no. 7074) and mouse (cat. no. 7076) were purchased from CST. Small interfering RNA (si)ATG5 was purchased from Shanghai GenePharma Co., Ltd., and Lipofectamine® 2000 was obtained from Thermo Fisher Scientific, Inc.
4
Brevilin A Cytotoxicity and Apoptosis Induction
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Brevilin A (Figure 1A ) was purchased from Dalian Meilun Biotechnology, Co., Ltd., and its purity was more than 98% as determined by HPLC. Temozolomide was obtained from Selleckchem (Munich, Germany). DMEM and FBS were obtained from Gibco (Eggenstein, Germany). Penicillin and streptomycin were purchased from Solarbio Co., Ltd. (Beijing, China). Annexin V–FITC/PI apoptosis detection kit, Reactive Oxygen Species (ROS) assay kit, mitochondrial membrane potential (MMP) assay kit with JC-1, MTT reagent, Western blotting reagents, and dimethyl sulfoxide (DMSO) were purchased from Beyotime Institute of Biotechnology (Haimen, Jiangsu, China). The glutathione (GSH) assay kit was obtained from Nanjing Jiancheng Bio-Engineering Institute (Nanjing, Jiancheng, China). The primary antibodies for cleaved forms of caspase-9, caspase-3, and PARP as well as p-JNK, JNK p-p38, and p38 were obtained from Cell Signaling Technology (Beverly, MA, USA) whereas primary antibodies for Bax, Bak, Bcl-2, Bcl-xL, Xiap, Cytochrome c, and GAPDH were obtained from Proteintech (Wuhan, China). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Sigma-Aldrich (St Louis, MO, USA).
5
BRDT Inhibitors for Cell Studies
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The BRDT inhibitors PLX51107 (cat. no. S8739) and INCB054329 (cat. no. S8753) were purchased from Selleck Chemicals. Both were dissolved in DMSO at 10 mmol/l, stored at −80°C and diluted in DMSO just before use. Exfect transfection reagent (cat. no. T202) was purchased from Vazyme Biotech Co., Ltd. Polyclonal antibody against BRDT (cat. no. NBP1-88357) was purchased from Novus Biologicals, Ltd. c-Myc (D3N8F; cat. no. 13987) and eIF4EBP1 (53H11; cat. no. 9644) rabbit monoclonal antibodies were both purchased from Cell Signaling Technology, Inc. Rabbit polyclonal antibody against β-actin (cat. no. 0407-3) was purchased from Huanan Biotechnology. DMSO was purchased from Sigma-Aldrich; Merck KGaA. NP40 lysates and western blotting reagents were all purchased from Beyotime Institute of Biotechnology.
6
Nanoparticle Synthesis and Cellular Assays
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The ZnO NPs (20 nm–50 nm, >98% pure) and ZnO used in the studies were acquired from Sigma-Aldrich (St. Louis, MO, USA), and were stored at 4 °C. Dimethyl-sulfoxide (DMSO), Fluo-3AM (fluorescent probe of Ca2+) and JC-1 (probe of mitochondrial membrane potential) were purchased from AMRESCO Inc. (Solon, OH, USA). A cell counting Kit-8 (CCK-8) and 2′,7′-dichlorofluorescensin diacetate (DCFH-DA) were obtained from Solarbio Science & Technology Co., Ltd. (Beijing, China). Cell culture reagents, including Dulbecco’s Modified Eagle’s medium (DMEM) and fetal bovine serum, were provided by Life Technologies (Therma Fisher Scientific, NY, USA) and Tianhang Co., Ltd. (Deqing, China). Western blotting reagents were acquired from the Beyotime Institute of Biotechnology (Haimen, China). An assay kit of oxidative stress species was obtained from Jiancheng Bioengineering Inc. (Jiangsu, China). The gluta solution was provided by the Aladdin Company (Shanghai, China). Sodium carboxymethylcellulose and Formalin Fix Solution (10%) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China).
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