A microtiter plate transglutaminase assay utilizing 5-(biotinamido)pentylamine as substrate

Role of transglutaminase 2 in PAC1 receptor mediated protection against hypoxia-induced cell death and neurite outgrowth in differentiating N2a neuroblastoma cells

“…The assay was performed as per the method described by Slaughter et al (1992) with the modifications of Lilley et al (1998). Briefly, 96-well microtitre plates were coated overnight at 4°C with 250 μl of N′,N′-dimethylcasein (10 mg ml -1 in 100 mM Tris-HCl, pH …”

A1 adenosine receptor-induced phosphorylation and modulation of transglutaminase 2 activity in H9c2 cells: A role in cell survival

“…Protein was determined using the bicinchoninic acid (BCA) protein assay [24], using a commercially available kit (Sigma-Aldrich Co. Ltd), with bovine serum albumin (BSA) as the standard Biotin-labeled cadaverine-incorporation assays were performed according to Slaughter et al. [25] with modifications [26], as described previously [27]. The biotin-labeled peptide cross-linking assay was performed according to the method of Trigwell et al [28] with minor modifications [27].…”

“…TGs can catalyse two types of cross-linking, namely (i) intra-, and/or inter-molecular covalent cross-links between protein-bound glutamine and protein-bound lysine residues, and (ii) cross-links between primary amines and protein-bound glutamine (both protein and polyamines links are transamidation).H9c2 cells were treated with CPA (1 µM) or adenosine (100 µM) for varying time periods and cell lysates subjected to the biotincadaverine amine-incorporation assay [25]. Both CPA and adenosine produced transient increases in TG2 catalysed biotin-cadaverine incorporation activity peaking at 10 min Figure 3).…”

Localization of Tissue Transglutaminase in Human Carotid and Coronary Artery Atherosclerosis: Implications for Plaque Stability and Progression

“…The lysates were centrifuged at 16,000 ϫ g for 30 minutes at 4°C, and the supernatant was used in an ELISA-based assay for transglutaminase, as previously described (Slaughter et al, 1992), with minimal modifications. This assay is based on the incorporation of the TG substrate 5-biotinamidopentylamine (5-BPA; Pierce Chemical Company) into immobilized N, NЈ-dimethylcasein, with subsequent detection using streptavidin-alkaline phosphatase.…”