Design, preparation and use of ligated phosphoproteins: A novel approach to study protein phosphatases by dot blot array, ELISA and Western blot assays

Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products

“…This enzyme can be widely found in human tissues, calf, shrimp as well as bacteria. Besides its effect on human health problem, ALP are usefully applied in various fields such as molecular research tool as well as enzyme immunoassays especially bacterial alkaline phosphatase (Dong & Zeikus, 1997;Sun et al, 2007). ), milk industry for validation of milk pasteurization (Rankin et al, 2010), cosmetics as a substance for the regeneration and metabolism of cells, environment for monitoring herbicides or pesticides (Muginova et al, 2007), clinical diagnosis as a marker and medicine such as calf intestinal alkaline phosphates, a therapeutic drug for lipopolysaccharide-mediated diseases (Beumer et al, 2003).…”

Extractive fermentation for enhanced production of alkaline phosphatase from Bacillus licheniformis MTCC 1483 using aqueous two-phase systems

“…The importance of alkaline phosphatase in clinical medicine and molecular biology has made it a popular subject for scientific study and commercial utility. Alkaline phosphatases have been extensively used in the diagnostics, immunology and molecular biology as biochemical markers in quantitative measurements of disease, linked enzymes in ELISA (Sun et al, 2007;Muginova et al, 2007;Baranov et al, 2008) and also used in nonradioactive detection techniques, probing, blotting and sequencing systems (Nilgiriwala et al, 2008;Sasajima et al, 2010). Alkaline phosphatase activity has been conventionally used as index of adequate pasteurization and the detection of alkaline phosphatase activity of thermally treated liquid milk products has become a common procedure for milk quality control (Rankin et al, 2010).…”

Phosphatase assays using ligated phosphoprotein substrates

“…Our work indicates that the use of ligated phosphoprotein substrates results in significantly higher sensitivity and consistency compared to the use of unligated peptides in ELISA (12). The IPL procedure is easy to do without sophisticated equipments.…”

“…Synthetic phosphopeptides were easily ligated through a carboxyl-terminal reactive thioester of an intein-generated carrier protein (CP) to yield the LPP substrates. These substrates were used for immobilization and ELISA analysis on a 96 well microtiter plates 12 . This approach circumvents the problems associated with inefficient and variable binding by small peptides to polystyrene microtiter plates, thus resulting in significantly higher sensitivity and consistency, when compared to the use of free peptide substrates.…”