| Assay Method Information | |
| | TG Activity Assays |
| Description: | For measurement of TG2 activity, the incorporation of biotinylated cadaverineinto immobilized N,N-dimethylcasein was used as described previously (Wanget al., 2012). For determination of the activity of TG1, TG3, TG6, and FXIIIa, commercial microassays were used (TG-CovTest; Covalab) (Hitomi et al., 2009), according to the manufacturer's instructions. For comparable purposes, a number ofTG2 assays were also undertaken using this assay. Briefly, TG-specific biotinylated peptides, including pepF11KA (FXIII pre-activated with thrombin),pepK5 (TG1), pepT26 (TG2), pepE51 (TG3), or pepY25 (TG6) (Hitomi et al., 2009; Fukui et al., 2013) were incubated with suitable TG family members in the presence of polyamine substrates immobilized onto 96-well microplates. The incorporated biotinylated peptides were measured using horseradish peroxidase-conjugated streptavidin and then measured using o-phenylenediamine dihydrochloride substrates. The absorbance was measured at 490 nm using a microplat |
| Affinity data for this assay | |
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