XTT Assay: A Colorimetric Assay to Assess Cell Viability and Mitochondrial Activity

Abstract

Trendowski, M., et al. Generation and Quantitative Analysis of Pulsed Low Frequency Ultrasound to Determine the Sonic Sensitivity of Untreated and Treated Neoplastic Cells. J. Vis. Exp. (2015)

This article describes the protocol for XTT assay which is a colorimetric assay to assess cell viability and mitochondrial activity.

Protocol

1. XTT Assay to Determine Cell Viability and Mitochondrial Activity of Treated U937 and THP1 Cells

1. Prior to the XTT assays, grow U937 and THP1 cells under the same conditions. Pretreat cells with the same concentration range of MeβCD for 30 min prior to sonication. Use the same ultrasound parameters as before except that cells are now sonicated using a range of 1-3 one sec pulses spaced one sec apart to develop a range of damage for the XTT kit to assess.
   NOTE: Many of these steps are taken directly from the XTT kit manual and are only repeated for the convenience of interested laboratories.
2. Collect cells by centrifug....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Iscove's Modified Dulbecco's Medium  w/ NaHCO3 & 25mM Hepes	Life Technologies	12440079
Branson SLPe 40kHz Cell Disruptor with  3" (25mm) Cuphorn	Branson Ultrasonics	101-063-726	sonication device
Tip One 100 μl and 1,000 μl Filter Tips	USA Scientific	1120-1840, 1126-7810
100 μl Micropipette	Wheaton	851164
1000 μl Micropipette	Wheaton	851168
Forma Scientific Dual chamber water jacketed Incubator	Forma Scientific	3131
XTT Cell Proliferation Assay Kit	ATCC	30-1011K
96-Well Microplate Reader	Cole-Palmer	EW-13055-54
U937 Human Monocytic Leukemia Cells	ATCC	CRL1593.2
THP1 Human Monocytic Leukemia Cells	ATCC	TIB-202