Activity and enantioselectivity of the enzymes was usually assayed by time-consuming chromatographic analysis of substrates and the corresponding reaction products.
The amounts of GCF MMP-8 were assayed by time-resolved IFMA (Medix Biochemica, Kauniainen, Finland) [ 45].
Cells were microinjected with cDNA encoding cyclin B1-Venus at a concentration of 3 ng/µl in G2 cells, using a semiautomatic microinjector (Eppendorf, Stevenage, UK) on a Leica DMIRBE microscope (Leica Microsystems, Milton Keynes, UK) and assayed by time-lapse DIC and fluorescence microscopy as previously described (Karlsson and Pines, 1998).
CMs from CSps (n = 20) and from CDCs (n = 20) were separately pooled and assayed by time-resolved immunofluorometric assay (TRIFA) before diafiltration on X-100A nonionic membranes (Diaflo, 43 mm diameter, Amicon) using a stirred cell (Model 52, Amicon); these CMs were finally concentrated to small volumes with 0.02 M Tris buffer solution, pH 7.2, 0.15 M NaCl.
RNA was then extracted and amplified from these cells, and assayed by real time PCR.
The correlation between genes assayed by real time PCR analysis and by microarray analysis was significant (R2 = .73, Figure 2A).Microarray analysis tended to underestimate the fold change, especially in the liver stage samples (Figure 2B).
Levels of Fas mRNA were assayed by real time RT-PCR.
In addition, twenty HTLV-I ELISA negative samples were assayed by real time PCR approach as negative controls.
Gene expression was assayed by Real Time PCR. Cell invasiveness was assayed through a Matrigel invasion assay; cell proliferation was determined through the soft agar assay, MTT, and [H] thymidine incorporation.
CBX5 RNA transcripts harvested from equivalent amounts of tumor and normal intestinal tissue of B6 Apc Min/+ mice heterozygous and homozygous for the insertion were assayed by real time PCR.
To assess the extent that exudate collected from the fruit suture vasculature might be contaminated a number of transcripts were assayed by real time RT-qPCR in both exudate and extracts from the surrounding, non-suture, pod tissue.
