Purification of alkaline phosphatase from chicken intestine by three-phase partitioning and use of phenyl-Sepharose 6B in the batch mode.
Alkaline phosphatase from chicken intestine was purified from the crude preparation employing three-phase partitioning and by the use of phenyl Sepharose-6B in the batch mode. TPP uses a combination of ammonium sulphate and t-butanol to precipitate proteins from crude aqueous extracts. The precipitated protein forms interface between lower aqueous phase and upper organic solvent phase. The fold purification of the finally purified enzyme was 80 and the activity recovery was 61%. The sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis of enzyme showed considerable purification and its molecular weight was found to be around 67 kDa.
Sharma
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Chemistry Department, Indian Institute of Technology, Delhi, India.
Sharma
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Gupta
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MN
D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000818:Animals; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D007422:Intestines / Q000201:enzymology*; D012685:Sepharose / Q000031:analogs & derivatives*
C020927:phenyl-sepharose; D012685:Sepharose; D000469:Alkaline Phosphatase