Plasma membrane calcium ATPase (PMCA4): A housekeeper for RT-PCR relative quantification of polytopic membrane proteins |
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Authors: | Anna Maria Calcagno Katherine J Chewning Chung-Pu Wu Suresh V Ambudkar |
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Institution: | (1) Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, DHHS, Bethesda, MD 20892–42546, USA |
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Abstract: | Background Although relative quantification of real-time RT-PCR data can provide valuable information, one limitation remains the selection
of an appropriate reference gene. No one gene has emerged as a universal reference gene and much debate surrounds some of
the more commonly used reference genes, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). At this time, no gene encoding
for a plasma membrane protein serves as a reference gene, and relative quantification of plasma membrane proteins is performed
with genes encoding soluble proteins, which differ greatly in quantity and in targeting and trafficking from plasma membrane
proteins. In this work, our aim was to identify a housekeeping gene, ideally one that codes for a plasma membrane protein,
whose expression remains the same regardless of drug treatment and across a wide range of tissues to be used for relative
quantification of real-time RT-PCR data for ATP binding cassette (ABC) plasma membrane transporters. |
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