Research reportImmunochemical analyses of human plasma fibronectin-cytosolic transglutaminase interactions
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Quantification of human tissue transglutaminase by a luminescence sandwich enzyme-linked immunosorbent assay
2011, Analytical BiochemistryCitation Excerpt :Nevertheless, our method showed a low background and low intra- and interassay variation. The analytical sensitivity of the new assay (40 pg tTG/ml) outranges those of other sandwich ELISAs with detection limits of 0.1 ng/ml [34], approximately 1 ng/ml [42], and 50 ng/ml [43,44]. The first of the above-mentioned assays, using a combination of mono- and polyclonal antibodies [34], was the most sensitive test until now.
A crucial sequence for transglutaminase type 2 extracellular trafficking in renal tubular epithelial cells lies in its N-terminal β-sandwich domain
2011, Journal of Biological ChemistryCitation Excerpt :Assay linearity was demonstrated by a TG2 standard curve produced by applying purified guinea pig liver TG2 to wells containing no cells (supplemental Fig. 1C). Extracellular TG2 antigen was measured using a modified quantitative ELISA adapted from Achyuthan et al. (31) based on the ability of TG2 to adhere to fibronectin. 8 × 104 cells/well were plated into a fibronectin-coated 96-well plate.
Validated sandwich ELISA for the quantification of tissue transglutaminase in tissue homogenates and cell lysates of multiple species
2008, Journal of Immunological MethodsExternal GTP-bound transglutaminase 2 is a molecular switch for chondrocyte hypertrophic differentiation and calcification
2005, Journal of Biological ChemistryCitation Excerpt :For qualitative evaluation of TG2 expression in conditioned media, transfected CH-8 cells were placed in serum-free medium for the last 8 h of incubation, and then the conditioned medium was precipitated as described above and separated by SDS-PAGE. TG2 was quantified in the conditioned medium after binding to a Nunc-Immuno Module plate (Nunc, Rochester, NY) for detection by direct enzyme-linked immunosorbent assay, using biotin-labeled TG2-specific antibody CUB7402 (38). To assess the ability of CH-8 cells overexpressing specific TG2 mutants to bind to fibronectin, Nunc-Immuno Module plates were coated with 0.5 μg/ml fibronectin, blocked with 3% bovine serum albumin, and incubated with aliquots of 25 μg in protein of cell lysates and detected as described (38).
Analysis of tissue transglutaminase function in the migration of Swiss 3T3 fibroblasts. The active-state conformation of the enzyme does not affect cell motility but is important for its secretion
2002, Journal of Biological ChemistryCitation Excerpt :The presence of tTGase in the protein pellet was detected by Western blotting as outlined above. Alternatively, the cell growth medium was lyophilized, reconstituted in one-tenth of the initial volume, and analyzed for tTGase antigen by a modified enzyme-linked immunosorbent assay (ELISA) method according to Achyuthan et al. (35) as described below. The protein concentration was determined according to the method of Lowry et al. (36).