Selection of reference genes for gene expression studies in human neutrophils by real-time PCR.
BACKGROUND
Reference genes, which are often referred to housekeeping genes, are frequently used to normalize mRNA levels between different samples. However the expression level of these genes may vary among tissues or cells, and may change under certain circumstances. Thus the selection of reference gene(s) is critical for gene expression studies. For this purpose, 10 commonly used housekeeping genes were investigated in isolated human neutrophils.


RESULTS
Initial screening of the expression pattern demonstrated that 3 of the 10 genes were expressed at very low levels in neutrophils and were excluded from further analysis. The range of expression stability of the other 7 genes was (from most stable to least stable): GNB2L1 (Guanine nucleotide binding protein, beta polypeptide 2-like 1), HPRT1 (Hypoxanthine phosphoribosyl transferase 1), RPL32 (ribosomal protein L32), ACTB (beta-actin), B2M (beta-2-microglobulin), GAPD (glyceraldehyde-3-phosphate dehydrogenase) and TBP (TATA-binding protein). Relative expression levels of the genes (from high to low) were: B2M, ACTB, GAPD, RPL32, GNB2L1, TBP, and HPRT1.


CONCLUSIONS
Our data suggest that GNB2L1, HPRT1, RPL32, ACTB, and B2M may be suitable reference genes in gene expression studies of neutrophils.
Zhang
Xiaozhu
X
The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, St, Paul's Hospital, University of British Columbia, Vancouver, Canada. xzhang@mrl.ubc.ca
Ding
Lily
L
Sandford
Andrew J
AJ
D000199:Actins / Q000032:analysis; D019204:GTP-Binding Proteins / Q000032:analysis; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling / Q000592:standards*; D005796:Genes; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000032:analysis; D009363:Neoplasm Proteins / Q000032:analysis; D009504:Neutrophils / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards*; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000032:analysis; D012015:Reference Standards; D012269:Ribosomal Proteins / Q000032:analysis; D001613:beta 2-Microglobulin / Q000032:analysis
D000199:Actins; D009363:Neoplasm Proteins; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D012269:Ribosomal Proteins; D001613:beta 2-Microglobulin; C041806:ribosomal protein L32; D012313:RNA; D007041:Hypoxanthine Phosphoribosyltransferase; D019204:GTP-Binding Proteins
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