Rao, N. M. ; Nagaraj, R. (1989) A recycling assay for alkaline phosphatase applied to studies on its transport in E. coli K12 Journal of Biochemical and Biophysical Methods, 19 (4). pp. 301-308. ISSN 0165-022X
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Official URL: http://linkinghub.elsevier.com/retrieve/pii/016502...
Related URL: http://dx.doi.org/10.1016/0165-022X(89)90061-4
Abstract
A recycling assay for alkaline phosphatase, based on its ability to hydrolyse NADP to NAD+, is presented. The product NAD+ is recycled in a coupled assay consisting of NADH regeneration and reduction of a nitroblue tetrazolium salt. This assay is 10-12 times more sensitive than the conventional assay. We demonstrate the role of energy poisons in transport of this protein into the periplasm by combining the improved detection with phase separation of the periplasmic and cytoplasmic alkaline phosphatase pools.
| Item Type: | Article |
|---|---|
| Source: | Copyright of this article belongs to Elsevier Science. |
| Keywords: | NADP; Coupled Assay; Respiratory Inhibitor; Protein Secretion |
| ID Code: | 23853 |
| Deposited On: | 01 Dec 2010 13:01 |
| Last Modified: | 09 Jun 2011 08:43 |
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