【MTT細胞增殖分析試劑盒】MTT Cell Proliferation and Cytotoxicity Assay Kit貨號abx090676

MTT細胞增殖分析試劑盒 | MTT Cell Proliferation and Cytotoxicity Assay Kit貨號abx090676

MTT Cell Proliferation and Cytotoxicity Assay Kit
MTT是一種檢測細胞增殖(cell proliferation)和細胞毒性(cytotoxicity)的試劑盒。 MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide] [3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化物]，一種黃色四唑(yellow tetrazole)，被粒線體脫氫酶(mitochondrial dehydrogenases)還原為不溶性紫色結晶甲臢。具有活性代謝的活細胞將MTT轉化為 formazan，然而，死細胞喪失了這種能力。因此，顏色形成僅作為活細胞的有用且方便的標記。測得的吸光度（570nm）與活細胞數成比例。高 O.D.s 表示細胞增殖，而低 O.D.s 表示細胞毒性。使用提供的 Formazan Diluent Buffer 時，無需去除固有的細胞培養基，因此可以避免不必要的測定錯誤。該檢測試劑盒為提供了easy-to-use，non-radioactive非放射性和high-throughput method 高通量的方法。評估細胞之 cell proliferation細胞增殖、 cell viability chemotaxis趨化性、cytotoxicity細胞毒性和 apoptosis細胞凋亡。

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MTT Staining Solution: 5 ml
Formazan Diluent Buffer: 55 ml

Non-radioactive and high-throughput method to measure cell proliferation/viability/cytotoxicity

Assay type: Quantitative
Detection method: Colorimetric
Platform: Microplate reader
Assay time: 3 hr 15 min
Sample type: Adherent cells, Suspension cells

Target	MTT Cell Proliferation and Cytotoxicity
Form	Liquid
Storage	Store the MTT Staining Solution at -20 °C in the dark for up to one year. Store the Formazan Diluent Buffer between 2-8°C for up to one year.
Directions for use	Protocol Bring all reagents to room temperature before use. Collect logarithmic phase cells and adjust the cell suspension concentration. Add 100 µl into each well of a cell-culture plate and adjust the cell density to 1000-10,000 cells per well (sterile PBS should be added to the edge of the wells of the plate). Seed cells in a 5% CO₂ incubator in the dark at 37 °C until cells are uniform at the bottom of the wells. The cell number for each well should be determined by cells’ size and proliferation rate. Add 0-10 µl of the compound of interest per well (for a concentration gradient, this can be added after cells adhere, or every 2 hours, or every half-day, etc). It is recommended to set 5 wells to analyse in duplicate. Incubate the cells for 16-48 hours, then observe under microscope by inverting the cells. Add 10 µl of MTT Staining Solution into each well. Culture for 4 hours. If the compound of interest can react with MTT, centrifuge and remove the cultured medium. Rinse with PBS carefully 2-3 times, then add the MTT-containing cultured medium. Add 100 µl of Formazan Diluent Buffer into each well, then mix gently at low speed for 10-30 min until the crystal is dissolved completely. If undissolved Formazan is still present, it is recommended to pipette up and down gently, avoiding bubbles, 2-3 times, or incubate overnight. Measure the O.D. value at 570 nm. Set blank wells (cell culture medium, MTT Staining Solution, Formazan diluent buffer) and control wells (cells, media containing the compound of interest with the same concentration, culture medium, MTT Staining Solution, Formazan diluent buffer). Notes: Ensure that the wells do not dry out due to evaporation, particularly when the cell culture time is long. Avoid using the outermost wells; use PBS, water or culture medium to replace any liquid that has evaporated; or place the 96-well plate near water in the incubator. The MTT Staining Solution should not be used if it has turned a gray-green colour. If a precipitate forms in the Formazan Diluent Buffer, heat using a 37 °C water bath until the precipitate dissolved. Incubation steps should be carried out in the dark. Wear Personal Protective Equipment (PPE), such as lab coats, lab glasses and gloves, when using this product.