housekeeping gene for B cell lines - (Jul/10/2008 )
Please can somebody answer me if i can use TBP gene as a reference in quantitative real time PCR for B cell lines.
well, nobody can answer you this because nobody knows your intention of the experiment (drug treatment, compare normal/malignant, compare stages of differentiation,...).
but you may want to look into this paper:
1: Anal Biochem. 2004 Jun 15;329(2):293-9.
Approach for defining endogenous reference genes in gene expression experiments.
García-Vallejo JJ, Van Het Hof B, Robben J, Van Wijk JA, Van Die I, Joziasse DH,
Van Dijk W.
Department of Molecular Cell Biology and Immunology, Vrije Universiteit Medical
Centre, 1081 BT, Amsterdam, The Netherlands.
The quantification of gene expression by real-time polymerase chain reaction
(PCR) has revolutionized the field of gene expression analysis. Due to its
sensitivity and flexibility it is becoming the method of choice for many
investigators. However, good normalization protocols still have to be implemented
to facilitate data exchange and comparison. We have designed primers for 10
unrelated genes and developed a simple protocol to detect genes with stable
expression that are suitable for use as endogenous reference genes for further
use in the normalization of gene expression data obtained by real-time PCR. Using
this protocol, we were able to identify human proteosome subunit Y as a reliable
endogenous reference gene for human umbilical vein endothelial cells treated for
up to 18 h with TNFalpha, IL-4, or IFNgamma and for B cells isolated from healthy
controls and patients suffering from IgA nephropathy. Other optional endogenous
reference genes that can be considered are phosphomannomutase (PPMM) and actin
for endothelial cells and glyceraldehyde-3-phosphate dehydrogenase and PPMM for B