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Edman Sequencing
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Protein Analytics - Edman Sequencing
 

PROTOCOLS
  • N-terminal sequence analysis is determined using Edman chemistry on an Applied Biosystems (AB) Procise liquid-pulse protein sequenator. PTH-Amino Acids are separated on a Brownlee C-18 reverse phase column (2.1 mm x 22 cm) at 55 C using a linear gradient of 14 to 44% buffer B over 17.6 min. Buffer A is 3.5% Tetrahydrofuran and 2% AB premix buffer concentrate. (The recipe for the concentrate is proprietary to AB, but they have published the approximate composition as follows: 10-20% Acetic Acid, < 10% Sodium Acetate, < 10% Sodium Hexanesulfonate, 65-75% Water.) Buffer B is 12% Isopropanol in Acetonitrile. PTH-amino acid analysis is performed by Model 610A software, version 2.1