MTT assay
Alexander Röntgen
Published: 2023-11-01 DOI: 10.17504/protocols.io.e6nvwdoqwlmk/v1
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Abstract
Protocol for MTT cytotoxicity assay on SH-SY5Y cells.
Steps
General
1.
Culture human SH-SY5Y cells at 37 °C and 5% CO2
Day 1
2.
Detach 80–90% confluent cells with Trypsin-EDTA
3.
Count cells and seed 10000 cells/well
4.
Incubate at 37 °C and 5% CO2 for 24 h
Day 2
5.
Aspirate medium from cells
6.
Add in protein solutions
7.
Place back at 37 °C and 5% CO2 for 24 h
Day 3
8.
Dilute MTT 1:10 in RPMI medium
9.
Aspirate medium from cells
10.
Add 100 µL MTT/RPMI solution per well
11.
Place back at 37 °C and 5% CO2 for 4 h
12.
Aspirate medium from cells
13.
Add 100 µL stop solution per well
14.
Incubate at 37 °C, 500 rpm on plate shaker
15.
Read out absorbance at 570 nm on plate reader
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