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Methods Mol Biol. 2018;1843:95-106. doi: 10.1007/978-1-4939-8704-7_8.

CRAC Channel Components Quantitative Expression (In Tissues and Cell Lines) Using qPCR.

Author information

1
INSERM U1003, Laboratoire d'Excellence Canaux Ioniques d'Intérêt Thérapeutique, Équipe Labellisée Par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
2
INSERM U1003, Laboratoire d'Excellence Canaux Ioniques d'Intérêt Thérapeutique, Équipe Labellisée Par la Ligue Nationale Contre le Cancer, SIRIC ONCOLille, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France. natacha.prevarskaya@univ-lille1.fr.

Abstract

Since last decade real-time qPCR has become a routine and robust approach for measuring the expression of genes of interest. Indeed, using qPCR, expression profile analyses are now possible and participate to the understanding of physiological or pathological role of channels such as calcium release-activated channels (CRAC). Initially discovered in lymphocyte T and immunity perturbations, recent studies have highlighted the role of CRAC channels in other pathologies such as cancer. Here we describe a protocol to quantify CRAC components expression, in tissue sample and cell lines, to validate knockdown strategies or identify their roles in physiological and pathological conditions (Hoth and Penner, J Physiol 465:359-386, 1993; Hoth and Penner, Nature 355:353-356, 1992; and Zweifach and Lewis, Proc Natl Acad Sci U S A 90:6295-6299, 1993).

KEYWORDS:

Expression profile; ORAI; STIM; qPCR

PMID:
30203280
DOI:
10.1007/978-1-4939-8704-7_8
[Indexed for MEDLINE]

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