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N-terminal Sequencing: Unlocking Protein Identity
N-terminal sequencing is a powerful technique that reveals the precise order of amino acids at the beginning of a protein chain. This information is crucial for understanding protein structure, function, and potential modifications.
The Process: Step by Step
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PITC Reaction: The protein's N-terminal amino acid reacts with PITC (phenylisothiocyanate), forming a stable complex.
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Cleavage with TFA: Trichloroacetic acid (TFA) selectively cleaves the first amino acid, still bound to PITC.
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Conversion to PTH: The PITC-amino acid is converted into a more stable PTH-amino acid for analysis.
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HPLC Identification: HPLC chromatography separates and identifies the PTH-amino acid by comparing it to known standards.
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Repeat: The process is repeated sequentially to determine the entire N-terminal sequence.
Why Choose N-terminal Sequencing?
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Protein Verification: Confirm the expected N-terminal sequence of a protein.
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Modification Detection: Identify post-translational modifications at the N-terminus.
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Proteolysis Studies: Map the sites where proteins are cleaved by enzymes.
Let Agile Bioscience guide you through the power of N-terminal sequencing – contact us to learn more!
N-Terminal Sequencing
