||ELISA, FACS, IF, IHC, IP, WB
||caspase-3 Antibody (E-8) is a mouse monoclonal IgG2a provided at 200 µg/ml raised against amino acids 1-277 representing the full length precursor form of caspase-3 of human origin recommended for detection of caspase-3 and full length procaspase-3 of human origin by WB, IP, IF, IHC(P), FCM and ELISA agarose conjugate for IP studies, sc-7272 AC, 500 µg/0.25 ml agarose HRP conjugate, sc-7272 HRP, 200 µg/ml fluorescein (sc-7272 fiTC) and rhodamine (sc-7272 TRITC) conjugates for immunofluorescence, 200 µg/ml phycoerythrin conjugate for intracellular FCM, sc-7272 PE, 100 tests Alexa Fluor® 405 (sc-7272 AF405), Alexa Fluor® 488 (sc-7272 AF488) and Alexa Fluor® 647 (sc-7272 AF647) conjugates for FCM or immunofluorescence; 100 µg/2 ml
||Caspase-3, also known as apopain, SCA-1, Yama and CPP32, is an aspartate-specific cysteine protease that belongs to the ICE subfamily of caspases. Caspase-3 is expressed in cells as an inactive precursor from which the p17 and p11 subunits of the mature caspase-3 are proteolytically generated during apoptosis. The caspase-3 precursor is first cleaved at Asp175-Ser176 to produce the p11 subunit and the p20 peptide. Subsequently, the p20 peptide is cleaved at Asp28-Ser29 to generate the mature p17 subunit. The active caspase-3 enzyme is a heterodimer composed of two p17 and two p11 subunits. At the onset of apoptosis, caspase-3 proteolytically cleaves PARP at an Asp216-Gly217 bond. During the execution of the apoptotic cascade, activated caspase-3 releases SREBP from the membrane of the ER in a proteolytic reaction that is distinct from their normal sterol-dependent activation. Caspase-3 cleaves and activates SREBPs between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Caspase-3 also cleaves and activates caspase-6, -7 and -9. The human caspase-3 gene encodes a cytoplasmic protein that is highly expressed in lung, spleen, heart, liver, kidney and cells of the immune system.
3 - Good (Publishable / Would use again)
Sample: Huh-7 cell lusate
Buffer: 20mM Tris-HCl, pH 7.5, 150mM NaCl, 2mM EDTA, 2mM EGTA, 0.5% (v/v) Triton X-100, 2mM dithiothreitol, protease inhibitor cocktail Complete (Roche), 10mM NaF, 1 mM okadaic acid, 1mM Na vanadate
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