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Edman Degradation N-terminal Sequence Analysis

The N-terminal sequence is closely related to the function and stability of proteins. Analyzing the N-terminal sequence of a protein can determine the starting point of the protein, which helps analyze the protein's advanced structure and reveal its biological function. Edman degradation involves the stepwise removal of amino acids from the N-terminus of a protein and identifying their types to determine the N-terminal amino acid sequence of a protein or polypeptide. The Edman sequencing reaction is a cyclic process, where the polypeptide/protein reacts with phenylisothiocyanate (PITC) under mild alkaline conditions, followed by acid treatment to release the terminal residue from the polypeptide chain as a phenylthiohydantoin derivative of the amino acid. This allows the identification of one amino acid per cycle, leading to the determination of the N-terminal amino acid sequence.

Diagram of Edman degradation N-terminal sequence analysis

BaiTai Packer Biotechnology BTP has established an N-terminal sequence analysis platform based on Shimadzu's Edman sequencing system and has obtained dual quality system certification of CNAS/ISO9001. We provide N-terminal sequencing services for purified protein products, antibodies, protein vaccines, recombinant collagen, and more for researchers and clients. Our sequencing system can determine the sequence information of 30 N-terminal amino acids. Using a specific protein loading system, it can determine 60-70 N-terminal amino acids.Feel free to contact us for more detailed information!

Laboratory Instruments

Fully Automatic Protein/Peptide Sequencer (PPSQ33A)

Technical Advantages

Samples can be prepared by gel electrophoresis transfer or membrane spotting without the need for any protein database, allowing direct analysis of amino acid information and accurate analysis of the N-terminal amino acid sequence.
Accurately distinguishes between isoleucine and leucine, glutamine and lysine.
Accurately identifies and characterizes truncated forms (N-1, N-2, etc.) in the full-length sequence.
Accurately determines the sequence of proteins containing a large number of repetitive fragments.

Case Illustration

To calibrate for 19 PTH-amino acids, we first test the mixed standard of 19 PTH amino acids. The calibration test chromatogram of the mixed standard is as follows:

Standard Calibration Test Chromatogram

After the calibration test of the PTH amino acid mixed standard is completed, the N-terminal sequence of the test sample is tested. The sample test chromatogram is as follows:

N-terminal Position 1

N-terminal Position 2

N-terminal Position 3

From the above chromatogram, the N-terminal sequence of the protein is determined to be Tyr-Val-Trp.

Bilingual Project Report in Chinese/English

In the technical report, BaiTai Packer provides you with a detailed bilingual report in Chinese and English, including:

1. Experimental procedures (in Chinese and English)
2. Relevant mass spectrometry parameters (in Chinese and English)
3. Detailed information on Edman degradation N-terminal sequence analysis
4. Mass spectrometry images
5. Raw data

Edman Degradation N-terminal Sequence AnalysisOne-stop Service

All you need to do is place an order and send the sample
BaiTai Packer provides a one-stop service: sample processing - on-machine analysis - data analysis - project report