Therefore, the N-terminal kit likely measures full-length betatrophin and the C-terminal kit measures total betatrophin, including both full-length protein and the C-terminal fragments.
Indeed, the betatrophin levels in lean and non-diabetic subjects determined by the C-terminal kit are higher than those determined by the N-terminal kit (Table 2).
Both full-length and total betatrophin concentrations were determined in serum samples by both N-terminal and C-terminal kits.
In conjunction with the Affymetrix GeneChip WT Terminal Labeling Kit, the Ambion WT Expression Kits were designed to generate amplified and biotinylated sense-strand DNA targets from the entire expressed genome and hybridized onto GeneChip Mouse Exon 1.0ST arrays (Affymetrix).
C-tails were added to the sheared DNA using the terminal transferase kit (New England Biolabs, MA) and the chain terminator ddCTP (GE Healthcare Biosciences, NJ).
The purified double strand cDNA served as a template to synthesize biotin-labeled cRNA using the GeneChip2 WT Terminal Labeling Kit (Affymetrix), and the biotin-labeled cRNA was purified using the GeneChip Sample Cleanup Module kit (Affymetrix) and fragmented to 35 200 bases.
Amplified DNA was fragmented and end-labeled using the GeneChip WT Double-Stranded DNA Terminal Labeling Kit, hybridized to GeneChip Human Promoter 1.0R arrays, and stained using the GeneChip Hybridization, Wash, and Stain Kit (all Affymetrix).
The DNA probes were 3'dig-ddUTP labelled using a terminal transferase kit (Roche).
Single-stranded cDNA was fragmented and end-labeled with the GeneChip WT Terminal Labeling Kit (Affymetrix).
The fragmented DNA was end-labeled using the GeneChip WT Double-Stranded DNA Terminal Labeling Kit (Affymetrix) according to the manufacturer's protocol.
This single-stranded DNA was labeled with biotinylated ddUTP using and Enzo BioArray Terminal Labeling kit (Enzo Life Sciences, Farmingdale, NY) according to the instructions provided by the manufacturer.
