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Monday, May 03, 2004
Description Cytokine-Induced Proliferation of Indicator Cell Lines Procedure Materials Indicator cell line (see Quick Guide Chart for a given cytokine) Culture Medium (RPMI-1640 supplemented with 10% FBS) 96-well flat-bottom culture plate (Costar Cat. No. 3595) MTT solution (Sigma Cat. No. M5655) 5 mg/ml stock in PBS kept at room temperature (protect from light) MTT Lysing Solution 20% SDS/50% DMF Instruments Pipettes and pipettors Humidified incubator 96-well micro test spectrophotometer Experiment Duration 24-48 hour incubation (see summary chart) 1 hour assay preparation Method Add 100 祃 of Culture Medium to each well of the 96-well Assay plate. Dilute samples and standard (see Quick Guide Chart for standard range) by 2-fold serial dilution in the Assay plate from row 2 to 12. Leave row 1 as blank. Wash indicator cells 3 times with RPMI-1640 and resuspend in Culture Medium at a density of 2-3.5 x 105 cells/ml (see Quick Guide Chart). Add 100 祃 of cell suspension to each well. Incubate cells for 24-48 hours (see Quick Guide Chart) at 37癈, 5% CO2 in a humidified incubator. Add 10 祃/well of 5 mg/ml MTT solution to the plate and incubate for 4 hours. Add 50 祃/well of MTT Lysing Solution to the plate and incubate overnight. Read plate at 570-650 nm. Graph standard curve and analyze data. Recipes Supplies Tips (责任编辑：泉水) |
Cytokine-Induced Proliferation of Indicator Cell Lines
时间:2006-07-24 14:10来源:Scienceboard.net 作者:admin
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