Benefits: Simple: The assay employs an add-mix-measure format combining a caspase-1 substrate, Z-WEHD-aminoluciferin, with a thermostable luciferase in an optimized, lytic reagent. Specific: Assay specificity for caspase-1 is confirmed by the subsequent use of the caspase-1 inhibitor Ac-YVAD-CHO. Direct Measurement: Measure caspase-1 activity directly in microplate wells or monitor released caspase-1 activity in culture medium from cells, preserving cells for use with other assays. See Results: Using this assay system, caspase-1 activation has been demonstrated in cell culture models of inflammasome activation, such as human THP-1 and mouse J774A.1 cell lines, and with primary cell models, such as human PBMCs. Detect Caspase-1 Activity in Cells or Released in Culture Medium 24,000 22,000 20,000 18,000 16,000 14,000 12,000 10,000 2,000 4,000 6,000 8,000 0 Pam3CSK4 Signal: background ratio 12.40 3.76 4.63 2.88 R848 culture medium Pam3CSK4 cells R848 no-cell control vehicle control treated cells treated cells + YVAD-CHO Inhibitor THP-1 cells were treated with two different inflammasome activators. Cells were either assayed directly or supernatant was transferred to another plate and assayed with or without a caspase-1 inhibitor to confirm specificity of the assay results. Learn More View the Poster Monitoring Inflammasome Activation with a Bioluminescent Caspase-1 Assay » enews | March 2016 Luminescence (RLU) 13125MA Pagina 4
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Promega Australia - Monthly Online Magazine - March 2016 Lees publicatie 10927Home