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Caspase 3 3g2 mouse mab

Manufactured by Cell Signaling Technology
Sourced in United States

Caspase-3 (3G2) mouse mAb is a monoclonal antibody that recognizes the cleaved form of caspase-3, a key effector of apoptosis. It is used for the detection of activated caspase-3 in various applications.

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3 protocols using caspase 3 3g2 mouse mab

1

Chromatographic Separation and MS Analysis

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Flash column chromatographic separations were accomplished on Biotage Isolera Four system (Biotage, Uppsala, Sweden). NMR was recorded on DRX-400 and DRX-500 NMR spectrometers (Bruker, Rheinstetten, Germany). ESI-, APCI- and HR ESI-MS were recorded on Agilent 1290 LC-MS or Agilent G6530 TOF MS spectrometer (Agilent Technologies Inc., California, USA). Western blot results were visualized on Mini-HD9-Auto Biomolecular imager (Leader Oriental Technology. LTD, Beijing, China). Dichlone and flavonoids were purchased from Fisher Scientific (Fair Lawn, NJ, USA) and Ark Pharm, Inc. (Arlington Heights, IL, USA). Cell lines were gotten from the American Type Culture Collection (Manassas, VA). Caspase-3 (3G2) mouse mAb, caspase-3 antibody, cleaved caspase-3 (Asp175) (5A1E) rabbit mAb, PARP (46D11) rabbit mAb and horseradish peroxidase (HRP) were purchased from Cell Signaling Technology, Inc (Danvers, MA, USA).
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2

Apoptosis Signaling Pathway Analysis

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SBC-3 cells (cell concentration: 5
× 105 cells/mL) were cultured in a six-well flat-bottom
plate and treated with EtOH/H2O (1:1), 10 μM cisplatin,
or 15 μM 1 for 24 h. The Western blotting analysis
was carried out by the same procedures as previously reported.26 (link) The following antibodies were recruited: β-actin
(8H10D10 Mouse mAb, product number 3700, 1:1000; Cell Signaling Technology,
Danvers, MA, USA), caspase-8 (1C12 Mouse mAb, product number 9746,
1:1000; Cell Signaling Technology), caspase-9 (C9 Mouse mAb, product
number 9508, 1:2000; Cell Signaling Technology), caspase-3 (3G2 Mouse
mAb, product number 9668, 1:1000, Cell Signaling Technology), PARP
(46D11 Rabbit mAb, product number 9532, 1:1000; Cell Signaling Technology),
Bax (2D2 Mouse mAb, product number 89477, 1:1000; Cell Signaling Technology),
Bcl-2 (124 Mouse mAb, product number 15071, 1:1000; Cell Signaling
Technology), anti-rabbit IgG, horseradish peroxidase (HRP)-linked
antibody (product number 7074, 1:10000; Cell Signaling Technology),
and anti-mouse IgG, HRP-linked antibody (product number 7076, 1:10000;
Cell Signaling Technology). The signals were detected using ECL Prime
Western Blotting Detection Reagents (GE Healthcare, Boston, MA, USA),
and photographed by a LAS-3000 luminescent image analyzer (FUJIFILM,
Tokyo, Japan).
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3

Immunodetection of Apoptotic Markers

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Protease Inhibitor Cocktail Set III was obtained from Calbiochem (Darmstadt, Germany); CD-31 (PECAm-1) was obtained from Invitrogen (Carlsbad, CA, USA) and Dulbecco's modified Eagle's medium (DMEM) and fetal calf serum (FCS) were obtained from Gibco-Invitrogen (Paisley, UK). Bovine serum albumin (BSA), streptavidin-horseradish peroxidase, 3,3' ,5,5'-tetramethylbenzidine (TmB) and poly-Llysine solution were obtained from Sigma-Aldrich (St. Louis, mO, USA). Taq DNA polymerase was obtained from Invitrogen. Oligonucleotide primers and Tris buffer were obtained from Sigma-Aldrich and dNTPs from Fermentas (Waltham, mA, USA). 3-3'-Diaminobenzidine chromogen/H 2 O 2 substrate in buffer solution (pH 7.5) (DAB kit) and rabbit anti-SRL polyclonal antisera were obtained from Bangalore Genei India Pvt. Ltd. (Bangalore India). Caspase-3 (3G2) mouse mAb was procured from Cell Signaling Technology, Inc. (Danvers, mA, USA). Species-specific HRP-labelled secondary antibodies were procured from Bio-Rad Laboratories (Hercules, CA, USA). Anticoagulated tubes were obtained from Greiener Bio-One (1 ml K3E K3EDTA; Frickenhausen, Germany) and a flat bottom 96-well micro-test plate were obtained from Tarsons (Kolkata, India). All other reagents were of analytical grade and milli-Q water was used for all the preparations.
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