Serial dilution of plasma revealed that a difference of 20%% can be detected by immunoblot and the subsequent quantification of the signals with OptiQuant software (not shown).
Variation in gene expression of large numbers of individuals is then assessed either directly by RNA-seq and subsequent quantification of reads mapped back to the assembled transcriptome (Smith et al. 2013) or by designing a custom microarray to quantify the expression of only a subset of genes contained in transcriptome (Renaut and Bernatchez 2011; Kvist et al. 2013).
RIP-chip, that is, immunoprecipitation of RNA-induced silencing complexes (RISC) with an Ago2-specific monoclonal antibody followed by RNA extraction and subsequent quantification of mRNAs on microarrays, has recently been utilized to identify mRNAs that are associated with the RNA-silencing machinery and are therefore targets of cellular miRNAs [ 35– 35].
Enzymatic digestion using GluC and trypsin generated chemically identical heavy and light peptide pairs, which were resolved by nano-LC-MS/MS, with subsequent quantification of acetylation stoichiometry in a site-specific manner.
Two approaches exist 1) primary autoantibody screening followed by quantification of risk by further autoantibody, genetic, and metabolic testing, and 2) primary genetic screening (e.g., of newborns using cord blood or dried blood spots) with determination of high-risk HLA (DR/DQ) gene alleles and subsequent quantification of risk by further autoantibody and metabolic testing.
The amount of acidic sites generated during oxidative treatment was quantified by reacting them with amine terminated octadecane and the subsequent quantification of the products of this grafting process by Fourier transform infrared spectroscopy.
Subsequent quantification of the nanostructure by fringe analysis indicates differences between the samples in terms of length, tortuosity, and separation of the graphitic fringes.
The decreased vascularization in treated tumors was further validated by immunostaining of corresponding tumor sections and subsequent quantification of CD31-positive area fractions (control group, 4.8 ± 0.50%; therapy group, 1.7 ± 0.37%; **P < 0.001; Figure 4D; representative pictures are shown in Figure 4E).
AC activity was tested using 30 min incubation of 0.02 μg sAC in 20 mM Tris/HCl pH 7.8, 5 mM ATP, 5 mM CaCl2 and 10 mM MgCl2 at 37°C, and subsequent quantification of cAMP formed by using an immunoassay kit (Assay Designs) as recommended by the manufacturer.
The remaining attached bacteria were heat-fixed by incubating at 60°C for 60 min. Subsequent quantification of biofilm was performed using the modified Christensen's method [41].
